Chimeric antigen receptors specific for g protein-coupled receptor class c group 5 member d (GPRC5d)

Abstract

To provide: chimeric antigen receptors (CARs), which contain antibody portions specific to G Protein-Coupled Receptor Class C Group 5 Member D (GPRC5D) and polynucleotides that encode CARs specific for GPRC5D; and genetically engineered cells containing such GPRC5D-binding receptors, and uses thereof in adoptive cell therapy.SOLUTION: A chimeric antigen receptor comprises: (1) an extracellular antigen-binding domain that specifically binds to GPRC5D, and comprises a variable heavy chain (VH) region and a variable light chain (VL) region which comprise specific amino acid sequences; (2) a spacer of at least 125 amino acids in length; (3) a transmembrane domain; and (4) an intracellular signaling region.SELECTED DRAWING: Figure 12C

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application is a continuation of U.S. Provisional Patent Application No. 62 / 754,576, filed November 1, 2018, entitled "CHIMERIC ANTIGEN RECEPTORS SPECIFIC FOR G PROTEIN-COUPLED RECEPTOR CLASS C GROUP 5 MEMBER D (GPRC5D)"; U.S. Provisional Patent Application No. 62 / 774,159, filed November 30, 2018, entitled "CHIMERIC ANTIGEN RECEPTORS SPECIFIC FOR G PROTEIN-COUPLED RECEPTOR CLASS C GROUP 5 MEMBER D (GPRC5D)"; This application claims priority to U.S. Provisional Patent Application No. 62 / 819,422, filed March 15, 2019, entitled "CHIMERIC ANTIGEN RECEPTORS SPECIFIC FOR G PROTEIN-COUPLED RECEPTOR CLASS C GROUP 5 MEMBER D (GPRC5D)"; U.S. Provisional Patent Application No. 62 / 904,197, filed September 23, 2019, entitled "CHIMERIC ANTIGEN RECEPTORS SPECIFIC FOR G PROTEIN-COUPLED RECEPTOR CLASS C GROUP 5 MEMBER D (GPRC5D)"; and U.S. Provisional Patent Application No. 62 / 904,187, filed September 23, 2019, entitled "BICISTRONIC POLYNUCLEOTIDE CONSTRUCTS ENCODING CHIMERIC ANTIGEN RECEPTORS," the contents of which are incorporated by reference in their entirety for all purposes.

[0002] INCORPORATION-BY-REFERENCE TO SEQUENCE LISTING This application is filed with an electronic Sequence Listing, which is provided in a file entitled 735042013740SeqList.TXT, created on October 31, 2019, and is 294 kilobytes in size. The information in the electronic Sequence Listing is incorporated by reference in its entirety.

[0003] Field In some aspects, the present disclosure relates to chimeric antigen receptors (CARs) comprising an antibody portion specific for G protein-coupled receptor class C group 5 member D (GPRC5D) and polynucleotides encoding CARs specific for GPRC5D. Additionally, the present disclosure relates to genetically engineered cells comprising such receptors that bind to GPRC5D and their use in adoptive cell therapy. [Background technology]

[0004] background G protein-coupled receptor class C group 5 member D (GPRC5D) is a G protein-coupled receptor whose specific function has not yet been determined. GPRC5D expression is high in bone marrow samples from patients with multiple myeloma (MM), whereas GPRC5D expression is minimal in bone marrow samples from patients with other hematopoietic malignancies. Based on its expression, GPRC5D could be a marker and therapeutic target for MM tumors. Various chimeric antigen receptors (CARs) that bind to GPRC5D and cells expressing such CARs are available. However, there remains a need for improved GPRC5D-binding CARs and modified GPRC5D-CAR-expressing targeting cells, for example, for use in adoptive cell therapy. Embodiments that meet this need are provided herein. Summary of the Invention

[0005] overview (1) an extracellular antigen-binding domain that specifically binds to human G protein-coupled receptor class C group 5 member D (GPRC5D), comprising: (i) a V as set forth in any of SEQ ID NOs: 21, 23, 25, 27, 29, 31, or 33; H heavy chain variable (V) domains comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the V domain amino acid sequence. H ) region; and (ii) a V region as set forth in any of SEQ ID NOs: 22, 24, 26, 28, 30, 32, or 34. L a light chain variable (V) region comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the V region amino acid sequence; L (1) an extracellular antigen-binding domain comprising a nucleotide sequence corresponding to the nucleotide sequence of ...

[0006] Also included is (1) an extracellular antigen-binding domain that specifically binds to human G protein-coupled receptor class C group 5 member D (GPRC5D), comprising: (i) a V selected from any one of SEQ ID NOs: 21, 23, 25, 27, 29, 31, or 33. H The heavy chain variable region (V) includes CDR-H1, CDR-H2, and CDR-H3 contained within the amino acid sequence of the region. H and (ii) V selected from any one of SEQ ID NOs: 22, 24, 26, 28, 30, 32, or 34. L The light chain variable (V) domain contains CDR-L1, CDR-L2, and CDR-L3. L(1) an extracellular antigen-binding domain comprising a nucleotide sequence corresponding to the nucleotide sequence of the target gene; (2) an extracellular antigen-binding domain comprising a nucleotide sequence corresponding to the target gene; (3) a spacer of at least 125 amino acids in length; (4) a transmembrane domain; and (5) an intracellular signaling region.

[0007] Also included is (1) an extracellular antigen-binding domain that specifically binds to human G protein-coupled receptor class C group 5 member D (GPRC5D), comprising: (i) a heavy chain complementarity-determining region 1 (CDR-H1) comprising an amino acid sequence selected from any one of SEQ ID NOs: 75, 78, 80, 82, 90, 93, 95, 97, 105, 108, 110, 112, 120, 123, 125, 127, 135, 138, 140, 142, 135, 152, 162, 165, 167, or 169; (b) a heavy chain complementarity-determining region 2 (CDR-H1) comprising an amino acid sequence selected from any one of SEQ ID NOs: 75, 78, 80, 82, 90, 93, 95, 97, 105, 108, 110, 112, 120, 123, 125, 127, 135, 138, 140, 142, 135, 152, 162, 165, 167, or 169; and (c) a heavy chain complementarity determining region 2 (CDR-H2) comprising an amino acid sequence selected from any one of SEQ ID NOs: 76, 79, 81, 83, 91, 94, 96, 98, 106, 109, 111, 113, 121, 124, 126, 128, 136, 139, 141, 143, 150, 153, 154, 155, 163, 166, 169, or 170; and (d) a heavy chain complementarity determining region 3 (CDR-H3) comprising an amino acid sequence selected from any one of SEQ ID NOs: 77, 84, 92, 99, 107, 114, 133, 129, 137, 144, 151, 156, 164, or 171. H and (ii) a light chain complementarity determining region 1 (CDR-L1) comprising an amino acid sequence selected from any one of SEQ ID NOs: 85, 88, 100, 103, 115, 118, 130, 133, 145, 148, 157, 160, 172, or 174; (b) a light chain complementarity determining region 2 (CDR-L2) comprising an amino acid sequence selected from any one of SEQ ID NOs: 86, 89, 101, 104, 116, 119, 131, 134, 146, 149, 158, or 161; and (c) a light chain complementarity determining region 3 (CDR-L3) comprising an amino acid sequence selected from any one of SEQ ID NOs: 87, 102, 117, 132, 147, 159, 173, 175, or 297. L(1) an extracellular antigen-binding domain comprising a nucleotide sequence corresponding to the nucleotide sequence of the target gene; (2) an extracellular antigen-binding domain comprising a nucleotide sequence corresponding to the target gene; (3) a spacer of at least 125 amino acids in length; (4) a transmembrane domain; and (5) an intracellular signaling region.

[0008] In some of any of the provided embodiments, the extracellular antigen-binding domain of the chimeric antigen receptor comprises (i) a V sequence as set forth in any of SEQ ID NOs: 21, 23, 25, 27, 29, 31, or 33. H heavy chain variable (V) domains comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the V domain amino acid sequence. H ) portion; and (ii) V as set forth in any of SEQ ID NOs: 22, 24, 26, 28, 30, 32 or 34. L a light chain variable (V) region comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the V region amino acid sequence; L ) area.

[0009] In some of any of the provided embodiments, the spacer is 125 or about 125 to 300 or about 300 amino acids in length, 125 or about 125 to 250 or about 250 amino acids in length, 125 or about 125 to 230 or about 230 amino acids in length, 125 or about 125 to 200 or about 200 amino acids in length, 125 or about 125 to 180 or about 180 amino acids in length, 125 or about 125 to 150 or about 150 amino acids in length, 150 or about 150 to 300 or about 300 amino acids in length, 150 or about 150 to 250 or about 250 amino acids in length, 150 or about 150 to 230 or about 230 amino acids in length, 150 or about 150 to 200 or about 200 amino acids in length, 150 Alternatively, the length may be about 150 to 180 or about 180 amino acids, 180 or about 180 to 300 or about 300 amino acids, 180 or about 180 to 250 or about 250 amino acids, 180 or about 180 to 230 or about 230 amino acids, 180 or about 180 to 200 or about 200 amino acids, 200 or about 200 to 300 or about 300 amino acids, 200 or about 200 to 250 or about 250 amino acids, 200 or about 200 to 230 or about 230 amino acids, 230 or about 230 to 300 or about 300 amino acids, 230 or about 230 to 250 or about 250 amino acids, or 250 or about 250 to 300 or about 300 amino acids.In some of any of the provided embodiments, the spacer is at least or at least about 130 amino acids in length, at least or at least about 140 amino acids in length, at least or at least about 150 amino acids in length, at least or at least about 160 amino acids in length, at least or at least about 17 ... 10 or about 170 amino acids in length, at least 180 or about 180 amino acids in length, at least 190 or about 190 amino acids in length, at least 200 or about 200 amino acids in length, at least 210 or about 210 amino acids in length, at least 220 or about 220 amino acids in length, at least 220 or about 220 amino acids in length, at least 221 or about 221 amino acids in length, at least 222 or about 222 amino acids in length, at least 223 or about 223 amino acids in length, at least 224 or about 224 amino acids in length, at least 225 or about 225 amino acids in length or is 225 or about 225 amino acids in length, at least 226 or at least about 226 amino acids in length, at least 227 or at least about 227 amino acids in length, at least 228 or at least about 228 amino acids in length, or at least 229 or at least about 229 amino acids in length, or 229 or about 229 amino acids in length, or has a length between any of the foregoing.

[0010] In some of any of the provided embodiments, the spacer is derived from an immunoglobulin. In some of any of the provided embodiments, the spacer comprises sequences of the hinge region, CH2 region, and CH3 region. In some of any of the provided embodiments, one or more of the hinge, CH2, and CH3 are derived in whole or in part from IgG4 or IgG2, optionally human IgG4 or human IgG2. In some of any of the provided embodiments, the hinge, CH2, and CH3 are derived from IgG4. In some of any of the provided embodiments, one or more of the hinge, CH2, and CH3 are chimeric and comprise sequences derived from IgG4 and IgG2. In some of any of the provided embodiments, the spacer comprises an IgG4 / 2 chimeric hinge, or a modified IgG4 comprising at least one amino acid substitution compared to human IgG4, an IgG2 / 4 chimeric CH2, and an IgG4 CH3 region.

[0011] In some of any of the provided embodiments, the spacer is or comprises: (i) the sequence set forth in SEQ ID NO:17; (ii) a functional variant of SEQ ID NO:17 having at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:17; or (iii) a contiguous portion of (i) or (ii) that is at least 125 amino acids in length. In some of any of the provided embodiments, the spacer is or comprises the sequence set forth in SEQ ID NO:17. In some of any of the provided embodiments, the spacer is or comprises the sequence encoded by the nucleotide sequence set forth in SEQ ID NO:48 (also set forth in SEQ ID NO:74).

[0012] In some of any of the embodiments provided, V H Area and V Lthe region comprises the amino acid sequence set forth in SEQ ID NOs: 21 and 22, respectively, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NOs: 21 and 22, respectively; H Area and V L the region comprises the amino acid sequence set forth in SEQ ID NOs: 23 and 24, respectively, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NOs: 23 and 24, respectively; H Area and V L the region comprises the amino acid sequence set forth in SEQ ID NOs: 25 and 26, respectively, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NOs: 25 and 26, respectively; H Area and V L the region comprises the amino acid sequence set forth in SEQ ID NOs: 27 and 28, respectively, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NOs: 27 and 28, respectively; H Area and V Lthe region comprises the amino acid sequence set forth in SEQ ID NOs: 29 and 30, respectively, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NOs: 29 and 30, respectively; H Area and V L the region comprises the amino acid sequence set forth in SEQ ID NOs: 31 and 32, respectively, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NOs: 31 and 32, respectively; or V H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NOs:33 and 34, respectively, or amino acid sequences having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NOs:33 and 34, respectively.

[0013] In some of any of the embodiments provided, V H The regions include CDR-H1, CDR-H2, and CDR-H3, each of which comprises the amino acid sequences of SEQ ID NOs: 80, 81, and 77, and V L The regions include CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences of SEQ ID NOs: 85, 86, and 87; H The regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 82, 83, and 84, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences of SEQ ID NOs: 88, 89, and 87; HThe regions include CDR-H1, CDR-H2, and CDR-H3, each of which contains the amino acid sequences of SEQ ID NOs: 95, 96, and 92, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences of SEQ ID NOs: 100, 101, and 102; H The regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 97, 98, and 99, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences of SEQ ID NOs: 103, 104, and 102; H The regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 110, 111, and 107, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences of SEQ ID NOs: 115, 116, and 117; H The regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 112, 113, and 114, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences of SEQ ID NOs: 118, 119, and 117; H The regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 125, 126, and 122, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences of SEQ ID NOs: 130, 131, and 132; H The regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 127, 128, and 129, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences of SEQ ID NOs: 133, 134, and 132; HThe regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 140, 141, and 137, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences of SEQ ID NOs: 145, 146, and 147; H The regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 142, 143, and 144, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences of SEQ ID NOs: 148, 149, and 147; H The regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 140, 154, and 151, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences of SEQ ID NOs: 157, 158, and 159; H The regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 142, 155, and 156, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences of SEQ ID NOs: 160, 161, and 159; H The regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 167, 168, and 164, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3, each containing the amino acid sequences of SEQ ID NOs: 172, 86, and 173; H The regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 169, 170, and 171, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 174, 89, and 175, respectively; or V HThe regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 169, 170, and 171, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3, which comprise the amino acid sequences of SEQ ID NOs: 174, 89, and 297, respectively.

[0014] In some of any of the embodiments provided, V H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NOs: 21 and 22, respectively; H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NOs: 23 and 24, respectively; H Area and V L The regions contain the amino acid sequences shown in SEQ ID NOs: 25 and 26, respectively; H Area and V L The regions include the amino acid sequences set forth in SEQ ID NOs: 27 and 28, respectively; H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NOs: 29 and 30, respectively; H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NOs: 31 and 32, respectively; or V H Area and V L The regions comprise the amino acid sequences shown in SEQ ID NOs: 33 and 34, respectively.

[0015] In some of any of the embodiments provided, V H Area and V Lthe region comprises the amino acid sequence set forth in SEQ ID NOs: 21 and 22, respectively, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NOs: 21 and 22, respectively; H Area and V L the region comprises the amino acid sequence set forth in SEQ ID NOs: 23 and 24, respectively, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NOs: 23 and 24, respectively; H Area and V L the region comprises the amino acid sequence set forth in SEQ ID NOs: 27 and 28, respectively, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NOs: 27 and 28, respectively; or V H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NOs:31 and 32, respectively, or amino acid sequences having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NOs:31 and 32, respectively.

[0016] In some of any of the embodiments provided, V H The regions include CDR-H1, CDR-H2, and CDR-H3, each of which comprises the amino acid sequences of SEQ ID NOs: 80, 81, and 77, and V LThe regions include CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences of SEQ ID NOs: 85, 86, and 87; H The regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 82, 83, and 84, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences of SEQ ID NOs: 88, 89, and 87; H The regions include CDR-H1, CDR-H2, and CDR-H3, each of which contains the amino acid sequences of SEQ ID NOs: 95, 96, and 92, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences of SEQ ID NOs: 100, 101, and 102; H The regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 97, 98, and 99, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences of SEQ ID NOs: 103, 104, and 102; H The regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 125, 126, and 122, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences of SEQ ID NOs: 130, 131, and 132; H The regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 127, 128, and 129, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences of SEQ ID NOs: 133, 134, and 132; H The regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 140, 154, and 151, respectively, and V LThe regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 157, 158, and 159, respectively; or V H The regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 142, 155, and 156, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3, which comprise the amino acid sequences of SEQ ID NOs: 160, 161, and 159, respectively.

[0017] In some of any of the embodiments provided, V H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NOs: 21 and 22, respectively; H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NOs: 23 and 24, respectively; H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NOs: 27 and 28, respectively; or V H Area and V L The regions comprise the amino acid sequences shown in SEQ ID NOs: 31 and 32, respectively.

[0018] In some of any of the provided embodiments, the extracellular antigen-binding domain is cross-reactive to or binds to mouse GPRC5D and / or cross-reactive to or binds to cynomolgus monkey GPRC5D. In some of any of the provided embodiments, the extracellular antigen-binding domain is not cross-reactive to or does not bind to mouse GPRC5D or cynomolgus monkey GPRC5D.

[0019] In some of any of the embodiments provided, V H Area and V LThe regions comprise the amino acid sequences set forth in SEQ ID NOs:27 and 28, respectively, or amino acid sequences having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NOs:27 and 28, respectively.

[0020] In some of any of the provided embodiments, the chimeric antigen receptor is V as set forth in SEQ ID NO:27. H The heavy chain variable region (V) includes CDR-H1, CDR-H2, and CDR-H3 contained within the amino acid sequence of the region. H ); and V as shown in SEQ ID NO:28 L The light chain variable (V) domain contains CDR-L1, CDR-L2, and CDR-L3. L In some of any of the provided embodiments, the V H The regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 125, 126, and 122, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 130, 131, and 132, respectively. H The regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 127, 128, and 129, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 133, 134, and 132, respectively. H The regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 120, 121, and 122, respectively, and V LThe regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 130, 131, and 132, respectively. H The regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 123, 124, and 122, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 130, 131, and 132, respectively. H Area and V L The regions comprise the amino acid sequences shown in SEQ ID NOs: 27 and 28, respectively.

[0021] In some of any of the provided embodiments, the extracellular antigen-binding domain is a single-chain antibody fragment. In some of any of the provided embodiments, the fragment is or comprises a single-chain variable fragment (scFv).

[0022] In some of any of the embodiments provided, V H Area and V L The regions are linked by a flexible linker. H Area and V L The region is the amino acid sequence In some of the embodiments provided, V is linked by a linker comprising: H Area and V L The regions are linked by a flexible linker. H Area and V L The region is the amino acid sequence Linked by a linker containing TIFF2025183309000003.tif4128.

[0023] In some of any of the embodiments provided, V H The area is V L and the amino acid sequence is amino-terminal to the region. In some of any of the provided embodiments, the antigen-binding domain comprises an amino acid sequence selected from any one of SEQ ID NOs: 1, 3, 5, 7, 9, 11, or 13, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence selected from any one of SEQ ID NOs: 1, 3, 5, 7, 9, 11, or 13. In some of any of the provided embodiments, the antigen-binding domain comprises an amino acid sequence selected from any one of SEQ ID NOs: 1, 3, 5, 7, 9, 11, or 13. In some of any of the provided embodiments, the antigen-binding domain comprises an amino acid sequence selected from any one of SEQ ID NOs: 1, 3, 7 or 11, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence selected from any one of SEQ ID NOs: 1, 3, 7 or 11. In some of any of the provided embodiments, the antigen-binding domain comprises an amino acid sequence selected from any one of SEQ ID NOs: 1, 3, 7 or 11. In some of any of the provided embodiments, the antigen-binding domain comprises the amino acid sequence set forth in SEQ ID NO:7, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:7. In some of any of the provided embodiments, the antigen-binding domain comprises the amino acid sequence set forth in SEQ ID NO:7.

[0024] In some of any of the embodiments provided, V H The area is V Land a region carboxy-terminal to the region. In some of any of the provided embodiments, the antigen-binding domain comprises an amino acid sequence selected from any one of SEQ ID NOs: 2, 4, 6, 8, 10, 12, or 14, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence selected from any one of SEQ ID NOs: 2, 4, 6, 8, 10, 12, or 14. In some of any of the provided embodiments, the antigen-binding domain comprises an amino acid sequence selected from any one of SEQ ID NOs: 2, 4, 6, 8, 10, 12, or 14. In some of any of the provided embodiments, the antigen-binding domain comprises an amino acid sequence selected from any one of SEQ ID NOs: 2, 4, 8 or 12, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence selected from any one of SEQ ID NOs: 2, 4, 8 or 12. In some of any of the provided embodiments, the antigen-binding domain comprises an amino acid sequence selected from any one of SEQ ID NOs: 2, 4, 8 or 12. In some of any of the provided embodiments, the antigen-binding domain comprises the amino acid sequence set forth in SEQ ID NO:8, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:8. In some of any of the provided embodiments, the antigen-binding domain comprises the amino acid sequence set forth in SEQ ID NO:8.In some of any of the embodiments provided, the antigen-binding domain is encoded by the nucleotide sequence shown in SEQ ID NO:264.

[0025] In some of any of the provided embodiments, the intracellular signaling region comprises an intracellular cytoplasmic signaling domain. In some of any of the provided embodiments, the intracellular signaling domain is capable of inducing a primary activation signal in a T cell, is a component of a T cell receptor (TCR), and / or comprises an immunoreceptor tyrosine-based activation motif (ITAM). In some of any of the provided embodiments, the intracellular signaling domain is or comprises the cytoplasmic signaling domain of the zeta chain of the CD3-zeta (CD3ζ) chain or a functional variant or signaling portion thereof.

[0026] In some of any of the provided embodiments, the intracellular signaling domain is a human intracellular signaling domain or is derived from a human protein. In some of any of the provided embodiments, the intracellular signaling domain is or comprises the sequence set forth in SEQ ID NO:20, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:20.

[0027] In some of any of the provided embodiments, the intracellular signaling region further comprises a costimulatory signaling region. In some of any of the provided embodiments, the costimulatory signaling region comprises the intracellular signaling domain of a T cell costimulatory molecule, or a signaling portion thereof. In some of any of the provided embodiments, the costimulatory signaling region comprises the intracellular signaling domain of CD28, 4-1BB, or ICOS, or a signaling portion thereof. In some of any of the provided embodiments, the costimulatory signaling region comprises the intracellular signaling domain of 4-1BB, or a signaling portion thereof. In some of any of the provided embodiments, the costimulatory signaling region is a human costimulatory signaling region or is derived from a human protein. In some of any of the provided embodiments, the costimulatory signaling region comprises the intracellular signaling domain of CD28, for example, the intracellular signaling domain of human CD28.

[0028] In some of any of the provided embodiments, the costimulatory signaling region is or comprises the sequence set forth in SEQ ID NO:46 or an amino acid sequence that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:46. In some of any of the provided embodiments, the costimulatory signaling region comprises the intracellular signaling domain of 4-1BB. In some of any of the provided embodiments, the costimulatory signaling region is or comprises the sequence set forth in SEQ ID NO:19 or an amino acid sequence that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:19.

[0029] In some of any of the provided embodiments, the costimulatory signaling region is between the transmembrane domain and the intracellular signaling region. In some of any of the provided embodiments, the transmembrane domain is or includes a transmembrane domain derived from CD4, CD28, or CD8. In some of any of the provided embodiments, the transmembrane domain is or includes a transmembrane domain derived from CD28. In some of any of the provided embodiments, the transmembrane domain is a human transmembrane domain or is derived from a human protein. In some of any of the provided embodiments, the transmembrane domain is or includes the sequence set forth in SEQ ID NO:18, or an amino acid sequence exhibiting at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:18.

[0030] Also, (1) an extracellular antigen-binding domain that specifically binds to a human G protein-coupled receptor (GPRC5D), (i) V as shown in SEQ ID NO:27 H a heavy chain variable region (V) comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the V region amino acid sequence; H and (ii) V as set forth in any of SEQ ID NO:28. L a light chain variable (V) region comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the V region amino acid sequence; L(1) an extracellular antigen-binding domain comprising a CD3-zeta (CD3ζ) chain region; (2) a spacer as set forth in SEQ ID NO:17; (3) a transmembrane domain derived from human CD28; and (4) an intracellular signaling region comprising the cytoplasmic signaling domain of the zeta chain of the CD3-zeta (CD3ζ) chain and the intracellular signaling domain of a T cell costimulatory molecule.

[0031] In some of any of the embodiments provided, V H The region is V shown in SEQ ID NO:27 H CDR-H1, CDR-H2, and CDR-H3 contained within the amino acid sequence of the region, and V L The region is V shown in SEQ ID NO:28 L or V H The regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 125, 126, and 122, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences of SEQ ID NOs: 130, 131, and 132; H The regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 127, 128, and 129, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences of SEQ ID NOs: 133, 134, and 132; H The regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 120, 121, and 122, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 130, 131, and 132, respectively; or V HThe regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 123, 124, and 122, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 130, 131, and 132, respectively. H The region is V shown in SEQ ID NO:27 H CDR-H1, CDR-H2, and CDR-H3 contained within the amino acid sequence of the region, and V L The region is V shown in SEQ ID NO:28 L or V H The regions include CDR-H1, CDR-H2, and CDR-H3, which contain the amino acid sequences of SEQ ID NOs: 125, 126, and 122, respectively, and V L The regions include CDR-L1, CDR-L2, and CDR-L3, which comprise the amino acid sequences of SEQ ID NOs: 130, 131, and 132, respectively.

[0032] Also, (1) an extracellular antigen-binding domain that specifically binds to a human G protein-coupled receptor (GPRC5D), which is represented by V as shown in SEQ ID NO:27. H Heavy chain variable (V) domains, including CDR-H1, CDR-H2, and CDR-H3, contained within the amino acid sequence H ) region; and the V L The light chain variable (V) domain contains CDR-L1, CDR-L2, and CDR-L3. L ) region; or a V region comprising CDR-H1, CDR-H2, and CDR-H3 containing the amino acid sequences of SEQ ID NOs: 125, 126, and 122, respectively. H V region, and CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 130, 131, and 132, respectively. LV region including CDR-H1, CDR-H2, and CDR-H3 containing the amino acid sequences of SEQ ID NOs: 127, 128, and 129, respectively H V region, and CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 133, 134, and 132, respectively. L V region including CDR-H1, CDR-H2, and CDR-H3 containing the amino acid sequences of SEQ ID NOs: 120, 121, and 122, respectively H V region, and CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 130, 131, and 132, respectively. L or V comprising CDR-H1, CDR-H2, and CDR-H3 comprising the amino acid sequences of SEQ ID NOs: 123, 124, and 122, respectively. H V region, and CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 130, 131, and 132, respectively. L (1) an extracellular antigen-binding domain comprising a region; (2) a spacer as set forth in SEQ ID NO:17; (3) a transmembrane domain derived from human CD28; and (4) an intracellular signaling region comprising the cytoplasmic signaling domain of the zeta chain of the human CD3-zeta (CD3ζ) chain and the intracellular signaling domain of human CD28 or human 4-1BB.

[0033] Also, (1) an extracellular antigen-binding domain that specifically binds to a human G protein-coupled receptor (GPRC5D), which is represented by V as shown in SEQ ID NO:27. H Heavy chain variable (V) domains, including CDR-H1, CDR-H2, and CDR-H3, contained within the amino acid sequence H ) region; and the V L The light chain variable (V) domain contains CDR-L1, CDR-L2, and CDR-L3. L) region; or V comprising CDR-H1, CDR-H2, and CDR-H3 comprising the amino acid sequences of SEQ ID NOs: 125, 126, and 122, respectively. H V region, and CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 130, 131, and 132, respectively. L (1) an extracellular antigen-binding domain that specifically binds to a human G protein-coupled receptor (GPRC5D), the V domain being represented by SEQ ID NO:27; (2) a spacer as set forth in SEQ ID NO:17; (3) a transmembrane domain derived from human CD28; and (4) an intracellular signaling region comprising the cytoplasmic signaling domain of the zeta chain of the human CD3-zeta (CD3ζ) chain and the intracellular signaling domain of human CD28. H Heavy chain variable (V) domains, including CDR-H1, CDR-H2, and CDR-H3, contained within the amino acid sequence H ) region; and the V L The light chain variable (V) domain contains CDR-L1, CDR-L2, and CDR-L3. L ) region; or V comprising CDR-H1, CDR-H2, and CDR-H3 comprising the amino acid sequences of SEQ ID NOs: 125, 126, and 122, respectively. H V region, and CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 130, 131, and 132, respectively. L (1) an extracellular antigen-binding domain comprising a region; (2) a spacer as set forth in SEQ ID NO:17; (3) a transmembrane domain derived from human CD28; and (4) an intracellular signaling region comprising the cytoplasmic signaling domain of the zeta chain of human CD3-zeta (CD3ζ) chain and the intracellular signaling domain of human 4-1BB.

[0034] In some of any of the provided embodiments, the extracellular antigen-binding domain comprises the VH The amino acid sequence of the region and V shown in SEQ ID NO:28 L and / or in some of any of the provided embodiments, the extracellular antigen-binding domain comprises the scFv set forth in SEQ ID NO:7 or SEQ ID NO:8. In some of any of the provided embodiments, the extracellular antigen-binding domain comprises the V set forth in SEQ ID NO:27. H The amino acid sequence of the region and V shown in SEQ ID NO:28 L and the extracellular antigen-binding domain comprises the scFv shown in SEQ ID NO: 7. In some of any of the provided embodiments, the extracellular antigen-binding domain comprises the V shown in SEQ ID NO: 27. H The amino acid sequence of the region and V shown in SEQ ID NO:28 L and the extracellular antigen-binding domain comprises the scFv shown in SEQ ID NO:8.

[0035] In some of any of the provided embodiments, the transmembrane domain is or comprises the sequence set forth in SEQ ID NO: 18, or an amino acid sequence that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 18. In some of any of the provided embodiments, the transmembrane domain is or comprises the sequence set forth in SEQ ID NO: 18.

[0036] In some of any of the provided embodiments, the intracellular signaling region comprises the sequence set forth in SEQ ID NO:20, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:20, and the sequence set forth in SEQ ID NO:46, or an amino acid sequence that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:46. In some of any of the provided embodiments, the intracellular signaling region is or comprises the sequence set forth in SEQ ID NO:20 and the sequence set forth in SEQ ID NO:46. In some of any of the provided embodiments, the intracellular signaling region comprises the sequence set forth in SEQ ID NO:20, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:20, and the sequence set forth in SEQ ID NO:19, or an amino acid sequence that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:19. In some of any of the provided embodiments, the intracellular signaling region is or comprises the sequence set forth in SEQ ID NO:20 and the sequence set forth in SEQ ID NO:19.

[0037] In some of any of the provided embodiments, the chimeric antigen receptor comprises, in order from its N-terminus to C-terminus, an antigen-binding domain, a spacer, a transmembrane domain, and an intracellular signaling region.

[0038] Also provided is a polynucleotide comprising a nucleotide sequence encoding any of the chimeric antigen receptors provided herein.

[0039] In some of any of the provided embodiments, the nucleic acid encoding the spacer comprises at least one modified splice donor site and / or modified splice acceptor site, wherein the modified splice donor site and / or modified splice acceptor site comprises one or more nucleotide modifications corresponding to a reference splice donor site and / or reference splice acceptor site contained in the sequence set forth in SEQ ID NO:73. In some of any of the provided embodiments, the one or more nucleotide modifications comprise an amino acid substitution. In some of any of the provided embodiments, the reference splice donor site and / or reference splice acceptor site is a canonical splice site, an atypical splice site, or a cryptic splice site. In some of any of the provided embodiments, the reference splice donor and / or reference splice acceptor site has a nucleotide sequence of at least 0.4, at least 0.5, at least 0.6, at least 0.70, at least 0.75, at least 0.80, at least 0.85, at least 0.90, at least 0.95, at least 0.96, at least 0.97, at least 0.98, at least 0.99, at least 10 ... 5, has a splice site prediction score of at least or about 0.99 or at least 1.0 or about 1.0; and / or the reference splice donor and / or reference splice acceptor site is predicted to be involved in a splice event at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99% or at least 100% of the time.

[0040] In some of any of the provided embodiments, the reference splice donor site has the sequence TIFF2025183309000004.tif11147; and / or the reference splice acceptor site comprises the sequence Includes TIFF2025183309000005.tif11128.

[0041] In some of any of the provided embodiments, the reference splice donor and / or reference splice acceptor site has a splice site prediction score of at least or about 0.70, at least 0.75 or about 0.75, at least 0.80 or about 0.80, at least 0.85 or about 0.85, at least 0.90 or about 0.90, at least 0.95 or about 0.95, at least 0.99 or about 0.99, or at least 1.0; and / or in some of any of the provided embodiments, the reference splice donor and / or reference splice acceptor site is predicted to be involved in a splice event at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or at least 100% of the time.

[0042] In some of any of the provided embodiments, the reference splice donor site has the sequence TIFF2025183309000006.tif4128; and / or the reference splice acceptor site is Contains TIFF2025183309000007.tif4128

[0043] In some of any of the provided embodiments, at least one of the one or more nucleotide modifications is present within 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 residues of a splice site junction of the reference splice acceptor and / or reference splice donor site.

[0044] In some of any of the provided embodiments, the one or more nucleotide modifications are silent and / or result in degenerate codons compared to SEQ ID NO:73 and / or do not change the amino acid sequence of the encoding spacer.

[0045] In some of any of the provided embodiments, the modified splice donor site is TIFF2025183309000008.tif11151; and / or modified splice acceptor sites are shown in In some of the embodiments provided, the modified splice donor site is TIFF2025183309000010.tif4128 and / or modified acceptor sites Shown in TIFF2025183309000011.tif11128.

[0046] In some of any of the provided embodiments, the spacer is encoded by the nucleotide sequence set forth in SEQ ID NO:74 (also set forth in SEQ ID NO:48), or a portion thereof. In some of any of the provided embodiments, the spacer is encoded by the nucleotide sequence set forth in SEQ ID NO:73, or a portion thereof. In some of any of the provided embodiments, the spacer is encoded by the nucleotide sequence set forth in SEQ ID NO:74, or a portion thereof. In some of any of the provided embodiments, the spacer is encoded by the nucleotide sequence set forth in SEQ ID NO:283, or a portion thereof. In some of any of the provided embodiments, the spacer is encoded by the nucleotide sequence set forth in SEQ ID NO:284, or a portion thereof. In some of any of the provided embodiments, the spacer is encoded by the nucleotide sequence set forth in SEQ ID NO:305, or a portion thereof.

[0047] In some of any of the provided embodiments, upon expression of the polynucleotide in a cell, RNA transcribed from the polynucleotide, optionally messenger RNA (mRNA), exhibits at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% RNA homogeneity.

[0048] In some of any of the provided embodiments, upon expression in a cell, RNA, optionally messenger RNA (mRNA), transcribed from the polynucleotide exhibits reduced heterogeneity compared to the heterogeneity of mRNA transcribed from a reference polynucleotide, wherein the reference polynucleotide encodes the same amino acid sequence as the polynucleotide but differs from the polynucleotide by the presence of one or more splice donor sites and / or one or more splice acceptor sites in the nucleic acid encoding the spacer, and / or comprises one or more nucleotide alterations compared to the polynucleotide, and / or comprises the spacer set forth in SEQ ID NO: 73. In some of any of the provided embodiments, the RNA heterogeneity is reduced by more than or about 10%, more than or about 15%, more than or about 20%, more than or about 25%, more than or about 30%, more than or about 40%, more than or about 50%, or more. In some of any of the provided embodiments, the RNA transcribed from the reference polynucleotide, optionally messenger RNA (mRNA), exhibits an RNA heterogeneity of greater than or about 10%, greater than or about 15%, greater than or about 20%, greater than or about 25%, greater than or about 30%, greater than or about 40%, greater than or about 40%, greater than or about 50%, or greater. In some of any of the provided embodiments, the RNA homogeneity and / or RNA heterogeneity is examined by agarose gel electrophoresis, chip-based capillary electrophoresis, analytical ultracentrifugation, field-flow fractionation, or liquid chromatography.

[0049] In some of any of the embodiments provided, the polynucleotide is codon-optimized for expression in human cells.

[0050] In some of any of the provided embodiments, the chimeric receptor is a first chimeric receptor, and the polynucleotide further comprises a nucleotide sequence encoding a second chimeric antigen receptor. Thus, also provided herein are polynucleotides encoding a first chimeric receptor that targets GPRC5D, such as any of those provided herein, and a second chimeric receptor. In some of any of the provided embodiments, the first chimeric receptor and the second chimeric receptor are separated by one or more multicistronic elements. In some of any of the provided embodiments, the one or more multicistronic elements are or comprise a ribosomal skipping sequence. In some embodiments, the ribosomal skipping sequence is a T2A, P2A, E2A, or F2A element. In some of any of the provided embodiments, the one or more multicistronic elements comprise the amino acid sequence set forth in SEQ ID NO: 37. In some of any of the provided embodiments, the one or more multicistronic elements are encoded by a nucleotide sequence selected from among SEQ ID NOs: 44, 45, and 319. In some of any of the provided embodiments, the nucleotide sequence encoding the one or more multicistronic elements is or comprises the sequence set forth in SEQ ID NO:319.

[0051] In some of any of the provided embodiments, the second chimeric receptor comprises an extracellular antigen-binding domain that specifically binds to a second antigen expressed on or associated with multiple myeloma, such as one other than GPRC5D. In some of any of the provided embodiments, the second CAR comprises an extracellular antigen-binding domain that binds to the second antigen, a spacer, a transmembrane domain, and an intracellular signaling region. In some of any of the provided embodiments, the second antigen is selected from B-cell maturation antigen (BCMA), CD38, CD138, CS-1, BAFF-R, TACI, or FcRH5. In some of any of the provided embodiments, the second antigen is BCMA.

[0052] In some of any of the provided embodiments, the second CAR comprises (1) an extracellular antigen-binding domain that specifically binds BCMA, the extracellular antigen-binding domain comprising (i) a V domain as set forth in any of SEQ ID NOs: 189, 191, 193, 195, or 197. H a heavy chain variable region (V) comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the V region amino acid sequence; H and (ii) V as set forth in any of SEQ ID NOs: 190, 192, 194, 196, or 198. L a light chain variable (V) region comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the V region amino acid sequence; L (1) an extracellular antigen-binding domain, including a V region; (2) a spacer; (3) a transmembrane region; and (4) an intracellular signaling region. In some of any of the provided embodiments, the V region of the second CAR H The region may be any of the V sequences shown in SEQ ID NOs: 189, 191, 193, 195 or 197. HV includes CDR-H1, CDR-H2, and CDR-H3 contained within the amino acid sequence of the region; L The region may be any of the V sequences shown in SEQ ID NOs: 190, 192, 194, 196, or 198. L The regions include CDR-L1, CDR-L2, and CDR-L3 contained within the amino acid sequence.

[0053] In some of any of the provided embodiments, the second CAR comprises (1) an extracellular antigen-binding domain that specifically binds to BCMA, the extracellular antigen-binding domain comprising (i) the V as set forth in SEQ ID NO:197. H a heavy chain variable region (V) comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the V region amino acid sequence; H ); and (ii) V as set forth in any of SEQ ID NO:198 L a light chain variable (V) region comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the V region amino acid sequence; L (1) an extracellular antigen-binding domain that specifically binds to BCMA, the extracellular antigen-binding domain comprising (i) a V region as set forth in SEQ ID NO:197; (2) a spacer; (3) a transmembrane domain; and (4) an intracellular signaling region. In some of any of the provided embodiments, the second CAR comprises (1) an extracellular antigen-binding domain that specifically binds to BCMA, the V region comprising (i) a V region as set forth in SEQ ID NO:197; H a heavy chain variable region (V) comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the V region amino acid sequence; H ); and (ii) V as set forth in any of SEQ ID NO:198 La light chain variable (V) region comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the V region amino acid sequence; L (1) an extracellular antigen-binding domain comprising a V region; (2) a spacer as set forth in SEQ ID NO: 17; (3) a transmembrane domain derived from human CD28; and (4) an intracellular signaling region comprising the cytoplasmic signaling domain of the zeta chain of the human CD3-zeta (CD3ζ) chain and the intracellular signaling domain of human 4-1BB. In some of any of the provided embodiments, the V of the second CAR H The region may be any of the V sequences shown in SEQ ID NO:197. H V includes CDR-H1, CDR-H2, and CDR-H3 contained within the amino acid sequence of the region; L The region may be any of the V sequences shown in SEQ ID NO:198. L In some of the provided embodiments, the second CAR is or comprises the amino acid sequence set forth in SEQ ID NO: 251. In some of the provided embodiments, the second CAR is encoded by the nucleotide sequence set forth in SEQ ID NO: 246.

[0054] In some of any of the provided embodiments, the second CAR comprises (1) a heavy chain variable region (V) that specifically binds to BCMA, the heavy chain variable region (V) comprising: (i) a heavy chain complementarity determining region 1 (CDR-H1) comprising an amino acid sequence selected from any one of SEQ ID NOs: 199, 202, 206, 209, 212, or 215; (b) a heavy chain complementarity determining region 2 (CDR-H2) comprising an amino acid sequence selected from any one of SEQ ID NOs: 200, 203, 207, 210, 213, or 216; and (c) a heavy chain complementarity determining region 3 (CDR-H3) comprising an amino acid sequence selected from any one of SEQ ID NOs: 201, 204, 205, 208, 211, 214, or 217. H and (ii) a light chain complementarity determining region 1 (CDR-L1) comprising an amino acid sequence selected from any one of SEQ ID NOs: 218, 221, 224, 227, 230, 233, or 235; (b) a light chain complementarity determining region 2 (CDR-L2) comprising an amino acid sequence selected from any one of SEQ ID NOs: 219, 222, 225, 228, 231, 234, or 236; and (c) a light chain complementarity determining region 3 (CDR-L3) comprising an amino acid sequence selected from any one of SEQ ID NOs: 220, 223, 226, 229, or 232. L (1) an extracellular antigen-binding domain, including a spacer region; (2) a transmembrane domain; and (3) an intracellular signaling region.

[0055] In some of any of the provided embodiments, the V of the second CAR H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 199, 200, and 201, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 218, 219, and 220, respectively; HThe regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 202, 203, and 204, respectively, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 221, 222, and 223, respectively; H The regions include CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 199, 200, and 205, respectively, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 224, 225, and 226, respectively; H The regions include CDR-H1, CDR-H2, and CDR-H3 containing the amino acid sequences of SEQ ID NOs: 206, 207, and 208, respectively, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 227, 228, and 229, respectively; H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 212, 213, and 214, respectively, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 233, 234, and 229, respectively; or the V of the second CAR H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 215, 216, and 217, and the V of the second CAR LThe regions include CDR-L1, CDR-L2, and CDR-L3, which comprise the amino acid sequences of SEQ ID NOs: 235, 236, and 232, respectively.

[0056] In some of any of the embodiments provided, the V of the encoded second CAR H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; or the V of the second CAR H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 215, 216, and 217, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 235, 236, and 232, respectively. In some of any of the provided embodiments, the V of the encoded second CAR H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively. In some of any of the provided embodiments, the V of the second CAR H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 215, 216, and 217, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3, which comprise the amino acid sequences of SEQ ID NOs: 235, 236, and 232, respectively.

[0057] In some of any of the embodiments provided, the V of the encoded second CARH Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NO: 189 and SEQ ID NO: 190, respectively, or an amino acid sequence that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 189 and SEQ ID NO: 190; H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NO: 191 and SEQ ID NO: 192, respectively, or an amino acid sequence that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 191 and SEQ ID NO: 192; H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NO: 193 and SEQ ID NO: 194, respectively, or an amino acid sequence that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 193 and SEQ ID NO: 194; H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NO: 195 and SEQ ID NO: 196, respectively, or an amino acid sequence that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 195 and SEQ ID NO: 196; or H Area and V LThe regions comprise the amino acid sequences set forth in SEQ ID NO:197 and SEQ ID NO:198, respectively, or amino acid sequences that exhibit at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:197 and SEQ ID NO:198.

[0058] In some of any of the embodiments provided, the V of the encoded second CAR H Area and V L The regions comprise the amino acid sequences shown in SEQ ID NO: 189 and SEQ ID NO: 190, respectively; H Area and V L The region comprises the amino acid sequence shown in SEQ ID NO: 191 and SEQ ID NO: 192; H Area and V L The region comprises the amino acid sequence shown in SEQ ID NO: 193 and SEQ ID NO: 194; H Area and V L The region comprises the amino acid sequence shown in SEQ ID NO: 195 and SEQ ID NO: 196; or the V of the second CAR H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NO: 197 and SEQ ID NO: 198, respectively. In some of any of the provided embodiments, the V of the encoded second CAR H Area and V L The regions comprise the amino acid sequences shown in SEQ ID NO:197 and SEQ ID NO:198, respectively.

[0059] In some of any of the provided embodiments, the extracellular antigen-binding domain of the encoded second CAR is a single-chain antibody fragment. In some of any of the provided embodiments, the encoded fragment is or comprises a single-chain variable fragment (scFv). In some of any of the provided embodiments, the V of the encoded second CAR is H Area and V L The regions are linked by a flexible linker. In some of any of the provided embodiments, the scFv of the encoded second CAR has the amino acid sequence In some of any of the provided embodiments, the encoded scFv of the second CAR comprises a linker comprising the amino acid sequence In some of the embodiments provided, the second CAR encoded by the V H The area is V L In some of the embodiments provided, the V H The area is V L It is carboxy-terminal to the region.

[0060] In some of any of the provided embodiments, the antigen-binding domain of the encoded second CAR comprises an amino acid sequence selected from any one of SEQ ID NOs: 227, 238, 239, 240, or 241, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence selected from any one of SEQ ID NOs: 227, 238, 239, 240, or 241. In some of any of the provided embodiments, the antigen-binding domain of the encoded second CAR comprises an amino acid sequence selected from any one of SEQ ID NOs: 227, 238, 239, 240, or 241. In some of any of the provided embodiments, the antigen-binding domain of the encoded second CAR comprises the amino acid sequence set forth in SEQ ID NO:241, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:241. In some of any of the provided embodiments, the antigen-binding domain of the encoded second CAR comprises the amino acid sequence set forth in SEQ ID NO:241.

[0061] In some of any of the provided embodiments, the V of the second CAR H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; or the V of the second CAR HThe regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 215, 216, and 217, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 235, 236, and 232, respectively; and / or the V of the second CAR H and / or the antigen-binding domain of the second CAR comprises the amino acid sequence set forth in SEQ ID NO:241, or a sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:241. In some of any of the provided embodiments, the VL region of the second CAR comprises the amino acid sequence set forth in SEQ ID NO:197 and SEQ ID NO:198, respectively. H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; and / or the V of the second CAR H and / or the antigen-binding domain of the second CAR comprises the amino acid sequence set forth in SEQ ID NO:241, or a sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:241. In some of any of the provided embodiments, the VL region of the second CAR comprises the amino acid sequence set forth in SEQ ID NO:197 and SEQ ID NO:198, respectively. HThe regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 215, 216, and 217, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 235, 236, and 232, respectively; and / or the V of the second CAR H the VL region and VL region comprise the amino acid sequences set forth in SEQ ID NO:197 and SEQ ID NO:198, respectively; and / or the antigen-binding domain of the second CAR comprises the amino acid sequence set forth in SEQ ID NO:241, or a sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:241.

[0062] In some of any of the provided embodiments, the transmembrane domain of the encoded second CAR is or comprises a transmembrane domain derived from CD4, CD28, or CD8, optionally a transmembrane domain derived from human CD4, human CD28, or human CD8.

[0063] In some of any of the provided embodiments, the transmembrane domain of the encoded second CAR is or comprises a transmembrane domain derived from human CD28 and / or is or comprises the sequence set forth in SEQ ID NO: 18, or an amino acid sequence exhibiting at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 18. In some of any of the provided embodiments, the transmembrane domain of the encoded second CAR is or comprises the sequence set forth in SEQ ID NO: 18.

[0064] In some of any of the provided embodiments, the intracellular signaling region of the encoded second CAR comprises an intracellular signaling domain. In some of any of the provided embodiments, the intracellular signaling domain of the encoded second CAR is capable of inducing a primary activation signal in a T cell, is a component of the T cell receptor (TCR), and / or comprises an immunoreceptor tyrosine-based activation motif (ITAM). In some of any of the provided embodiments, the intracellular signaling domain of the encoded second CAR is or comprises the cytoplasmic signaling domain of the zeta chain of the CD3-zeta (CD3ζ) chain or a functional variant or signaling portion thereof, optionally the cytoplasmic signaling domain of the human CD3 zeta chain. In some of any of the provided embodiments, the intracellular signaling region of the encoded second CAR comprises the sequence set forth in SEQ ID NO:20, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:20.

[0065] In some of any of the provided embodiments, the intracellular signaling region of the encoded second CAR further comprises a costimulatory signaling region. In some of any of the provided embodiments, the costimulatory signaling region of the encoded second CAR comprises the intracellular signaling domain of a T cell costimulatory molecule or a signaling portion thereof. In some of any of the provided embodiments, the costimulatory signaling region of the encoded second CAR comprises the intracellular signaling domain of CD28, 4-1BB, or ICOS, or a signaling portion thereof, optionally the intracellular signaling domain of human CD28, human 4-1BB, or human ICOS. In some of any of the provided embodiments, the costimulatory signaling region of the encoded second CAR comprises the intracellular signaling domain of 4-1BB or a signaling portion thereof. In some of any of the provided embodiments, at least one of the first chimeric antigen receptor and the second chimeric antigen receptor comprises an intracellular signaling region comprising the intracellular signaling domain of 4-1BB or a signaling portion thereof, optionally the intracellular signaling domain of human 4-1BB.

[0066] In some of any of the provided embodiments, the costimulatory signaling region of the encoded second CAR comprises the intracellular signaling domain of human CD28; and / or the sequence set forth in SEQ ID NO:46, or an amino acid sequence that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:46.

[0067] In some of any of the provided embodiments, the costimulatory signaling region of the encoded second CAR comprises the intracellular signaling domain of human 4-1BB; and / or the sequence set forth in SEQ ID NO:19, or an amino acid sequence that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:19.

[0068] In some of any of the embodiments provided, the encoded second chimeric antigen receptor comprises, in order from its N-terminus to C-terminus, an antigen-binding domain, a spacer, a transmembrane domain, and an intracellular signaling region.

[0069] In some of any of the provided embodiments, at least one of the polynucleotide sequence encoding the first chimeric antigen receptor and the polynucleotide sequence encoding the second chimeric antigen receptor is a codon-divergent form. In some of any of the provided embodiments, the polynucleotide sequence encoding the first chimeric antigen receptor and the polynucleotide sequence encoding the second chimeric antigen receptor have no more than about 30, no more than about 20, or no more than about 10 contiguous base pairs of sequence identity.

[0070] In some of any of the provided embodiments, the nucleotide sequence encoding the CAR is operably linked to a promoter to control expression of the encoded CAR when the polynucleotide is expressed in a cell into which it has been introduced, and optionally the promoter is a heterologous promoter, and optionally the heterologous promoter is or comprises the human elongation factor 1 alpha (EF1α) promoter or MND promoter, or a variant thereof.

[0071] In some of any of the provided embodiments where the polynucleotide encodes two CARs, the nucleotide sequence encoding the first CAR is operably linked to a first promoter for controlling expression of the first CAR when expressed in a cell into which the polynucleotide is introduced, and the nucleotide sequence encoding the second CAR is operably linked to a second promoter for controlling expression of the second CAR when expressed in a cell into which the polynucleotide is introduced. In some embodiments, the first promoter and the second promoter are independently heterologous promoters, for example, where the heterologous promoter is or comprises a human elongation factor 1 alpha (EF1α) promoter or an MND promoter, or a variant thereof. In some such embodiments, the first promoter and the second promoter are the same. In some such embodiments, the first promoter and the second promoter are different.

[0072] Also provided are vectors comprising any of the provided polynucleotides. In some of any of the provided embodiments, the vector is a viral vector. In some of any of the provided embodiments, the viral vector is a lentiviral vector or a retroviral vector.

[0073] Also provided are modified cells comprising any of the chimeric antigen receptors provided herein. In some of any of the provided embodiments, the modified cells comprise a chimeric antigen receptor provided herein and further comprise a polynucleotide comprising a nucleotide sequence encoding a second chimeric antigen receptor.

[0074] Also provided are modified cells comprising any of the polynucleotides provided herein.

[0075] In some of any of the provided embodiments, the second chimeric receptor of the provided cell comprises an extracellular antigen-binding domain that specifically binds to a second antigen expressed on or associated with multiple myeloma. In some of any of the provided embodiments, the second CAR comprises an extracellular antigen-binding domain that binds to the second antigen, a spacer, a transmembrane domain, and an intracellular signaling region. In some of any of such embodiments, the second antigen is selected from B-cell maturation antigen (BCMA), CD38, CD138, CS-1, BAFF-R, TACI, or FcRH5. In some of any of the provided embodiments, the second antigen is BCMA.

[0076] In some of any of the embodiments of the modified cells provided, the second CAR comprises (1) an extracellular antigen-binding domain that specifically binds BCMA, the extracellular antigen-binding domain comprising (i) a V domain as set forth in any of SEQ ID NOs: 189, 191, 193, 195, or 197. H a heavy chain variable region (V) comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the V region amino acid sequence; H and (ii) V as set forth in any of SEQ ID NOs: 190, 192, 194, 196, or 198. L a light chain variable (V) region comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the V region amino acid sequence; L (1) an extracellular antigen-binding domain comprising a V region; (2) a spacer; (3) a transmembrane region; and (4) an intracellular signaling region. H The region may be any of the V sequences shown in SEQ ID NOs: 189, 191, 193, 195 or 197. HV includes CDR-H1, CDR-H2, and CDR-H3 contained within the amino acid sequence of the region; L The region may be any of the V sequences shown in SEQ ID NOs: 190, 192, 194, 196, or 198. L The regions include CDR-L1, CDR-L2, and CDR-L3 contained within the amino acid sequence.

[0077] In some of any of the embodiments of the provided engineered cells, the second CAR comprises (1) a heavy chain variable region (V) that specifically binds to BCMA, the VV comprising: (i) a heavy chain complementarity determining region 1 (CDR-H1) comprising an amino acid sequence selected from any one of SEQ ID NOs: 199, 202, 206, 209, 212, or 215; (b) a heavy chain complementarity determining region 2 (CDR-H2) comprising an amino acid sequence selected from any one of SEQ ID NOs: 200, 203, 207, 210, 213, or 216; and (c) a heavy chain complementarity determining region 3 (CDR-H3) comprising an amino acid sequence selected from any one of SEQ ID NOs: 201, 204, 205, 208, 211, 214, or 217. H and (ii) a light chain complementarity determining region 1 (CDR-L1) comprising an amino acid sequence selected from any one of SEQ ID NOs: 218, 221, 224, 227, 230, 233, or 235; (b) a light chain complementarity determining region 2 (CDR-L2) comprising an amino acid sequence selected from any one of SEQ ID NOs: 219, 222, 225, 228, 231, 234, or 236; and (c) a light chain complementarity determining region 3 (CDR-L3) comprising an amino acid sequence selected from any one of SEQ ID NOs: 220, 223, 226, 229, or 232. L (1) an extracellular antigen-binding domain, including a spacer region; (2) a transmembrane domain; and (3) an intracellular signaling region.

[0078] In some of any of the embodiments of the modified cells provided, the V of the second CAR HThe regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 199, 200, and 201, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 218, 219, and 220, respectively; H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 202, 203, and 204, respectively, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 221, 222, and 223, respectively; H The regions include CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 199, 200, and 205, respectively, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 224, 225, and 226, respectively; H The regions include CDR-H1, CDR-H2, and CDR-H3 containing the amino acid sequences of SEQ ID NOs: 206, 207, and 208, respectively, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 227, 228, and 229, respectively; H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 212, 213, and 214, respectively, and the V of the second CAR LThe regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 233, 234, and 229, respectively; or the V of the second CAR H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 215, 216, and 217, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3, which comprise the amino acid sequences of SEQ ID NOs: 235, 236, and 232, respectively.

[0079] In some of any of the embodiments of the modified cells provided, the V of the second CAR H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; or the V of the second CAR H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 215, 216, and 217, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3, which comprise the amino acid sequences of SEQ ID NOs: 235, 236, and 232, respectively.

[0080] In some of any of the embodiments of the modified cells provided, the V of the second CAR H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NO: 189 and SEQ ID NO: 190, respectively, or an amino acid sequence that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 189 and SEQ ID NO: 190;H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NO: 191 and SEQ ID NO: 192, respectively, or an amino acid sequence that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 191 and SEQ ID NO: 192; H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NO: 193 and SEQ ID NO: 194, respectively, or an amino acid sequence that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 193 and SEQ ID NO: 194; H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NO: 195 and SEQ ID NO: 196, respectively, or an amino acid sequence that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 195 and SEQ ID NO: 196; or H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NO:197 and SEQ ID NO:198, respectively, or amino acid sequences that exhibit at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:197 and SEQ ID NO:198.

[0081] In some of any of the embodiments of the modified cells provided, the V of the second CAR H Area and V L The regions comprise the amino acid sequences shown in SEQ ID NO: 189 and SEQ ID NO: 190, respectively; H Area and V L The region comprises the amino acid sequence shown in SEQ ID NO: 191 and SEQ ID NO: 192; H Area and V L The region comprises the amino acid sequence shown in SEQ ID NO: 193 and SEQ ID NO: 194; H Area and V L The region comprises the amino acid sequence shown in SEQ ID NO: 195 and SEQ ID NO: 196; or the V of the second CAR H Area and V L The regions comprise the amino acid sequences shown in SEQ ID NO:197 and SEQ ID NO:198, respectively.

[0082] In some of the embodiments of the modified cells provided, the extracellular antigen-binding domain of the second CAR is a single-chain antibody fragment. In some of such embodiments, the fragment is or comprises a single-chain variable fragment (scFv).

[0083] In some of any of the embodiments of the modified cells provided, the V of the second CAR H Area and V L The regions are connected by a flexible linker. In some such embodiments, the linker has the amino acid sequence TIFF2025183309000014.tif4128. In some such embodiments, the linker comprises the amino acid sequence Includes TIFF2025183309000015.tif4128.

[0084] In some of any of the embodiments of the modified cells provided, V H The area is VL In some of the embodiments of the modified cells provided, the V H The area is V L In some of the embodiments of the modified cells provided, the antigen-binding domain of the second CAR comprises an amino acid sequence selected from any one of SEQ ID NOs: 227, 238, 239, 240, or 241, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence selected from any one of SEQ ID NOs: 227, 238, 239, 240, or 241.

[0085] In some of the embodiments of the modified cells provided, the antigen-binding domain of the second CAR comprises an amino acid sequence selected from any one of SEQ ID NOs: 227, 238, 239, 240, or 241. In some of the embodiments of the modified cells provided, the V of the second CAR comprises an amino acid sequence selected from any one of SEQ ID NOs: 227, 238, 239, 240, or 241. H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; or the V of the second CAR H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 215, 216, and 217, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 235, 236, and 232, respectively; and / or the V of the second CAR Hthe VL region and VL region comprise the amino acid sequences set forth in SEQ ID NO:197 and SEQ ID NO:198, respectively; and / or the antigen-binding domain of the second CAR comprises the amino acid sequence set forth in SEQ ID NO:241, or a sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:241.

[0086] In some of the embodiments of the modified cells provided, the antigen-binding domain of the second CAR comprises the amino acid sequence set forth in SEQ ID NO: 241. In some of the embodiments of the modified cells provided, the V of the second CAR comprises the amino acid sequence set forth in SEQ ID NO: 241. H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; and / or the V of the second CAR H the VL region and VL region comprise the amino acid sequences set forth in SEQ ID NO:197 and SEQ ID NO:198, respectively; and / or the antigen-binding domain of the second CAR comprises the amino acid sequence set forth in SEQ ID NO:241, or a sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:241.

[0087] In some of the embodiments of the modified cells provided, the transmembrane domain of the second CAR is or comprises a transmembrane domain derived from CD4, CD28, or CD8, optionally a transmembrane domain derived from human CD4, human CD28, or human CD8. In some of the embodiments of the modified cells provided, the transmembrane domain of the second CAR is or comprises a transmembrane domain derived from human CD28; and / or the transmembrane domain is or comprises the sequence set forth in SEQ ID NO: 18, or an amino acid sequence exhibiting at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 18. In some of the embodiments of the modified cells provided, the transmembrane domain of the second CAR is or comprises the sequence set forth in SEQ ID NO: 18.

[0088] In some of any of the embodiments of the modified cells provided, the intracellular signaling region of the second CAR comprises an intracellular signaling domain.

[0089] In some of the embodiments of any of the provided modified cells, the intracellular signaling domain of the second CAR is capable of inducing a primary activation signal in a T cell and is a component of a T cell receptor (TCR) and / or comprises an immunoreceptor tyrosine-based activation motif (ITAM). In some of the embodiments of any of the provided modified cells, the intracellular signaling domain of the second CAR is or comprises the cytoplasmic signaling domain of the zeta chain of the CD3-zeta (CD3ζ) chain or a functional variant or signaling portion thereof, optionally the cytoplasmic signaling domain of the human CD3 zeta chain. In some of the embodiments of any of the provided modified cells, the intracellular signaling region of the second CAR comprises the sequence set forth in SEQ ID NO:20, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:20.

[0090] In some of the embodiments of any of the provided modified cells, the intracellular signaling region of the second CAR further comprises a costimulatory signaling region. In some of the embodiments of any of the provided modified cells, the costimulatory signaling region of the second CAR comprises the intracellular signaling domain of a T cell costimulatory molecule or a signaling portion thereof. In some of the embodiments of any of the provided modified cells, the costimulatory signaling region of the second CAR comprises the intracellular signaling domain of CD28, 4-1BB, or ICOS, or a signaling portion thereof, optionally the intracellular signaling domain of human CD28, human 4-1BB, or human ICOS. In some of the embodiments of any of the provided modified cells, the costimulatory signaling region of the second CAR comprises the intracellular signaling domain of 4-1BB or a signaling portion thereof. In some of the embodiments of any of the provided modified cells, at least one of the first chimeric antigen receptor and the second chimeric antigen receptor comprises an intracellular signaling region comprising the intracellular signaling domain of 4-1BB or a signaling portion thereof, optionally the intracellular signaling domain of human 4-1BB.

[0091] In some of any of the embodiments of the modified cells provided, the costimulatory signaling region of the second CAR comprises the intracellular signaling domain of human CD28; and / or the sequence set forth in SEQ ID NO:46, or an amino acid sequence exhibiting at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:46.

[0092] In some of any of the embodiments of the modified cells provided, the costimulatory signaling region of the second CAR comprises the intracellular signaling domain of human 4-1BB; and / or the sequence set forth in SEQ ID NO:19, or an amino acid sequence that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:19.

[0093] In some of any of the embodiments of the modified cells provided, the encoded second chimeric antigen receptor comprises, in N- to C-terminal order, an antigen-binding domain, a spacer, a transmembrane domain, and an intracellular signaling region.

[0094] In some of any of the provided embodiments, the modified cells are lymphocytes. In some of any of the provided embodiments, the modified cells are NK cells or T cells. In some of any of the provided embodiments, the modified cells are T cells, and the T cells are CD4+ T cells or CD8+ T cells.

[0095] In some of any of the provided embodiments, the modified cells are modified primary cells obtained from the subject.

[0096] In some of any of the provided embodiments, the modified cells, of a plurality of said modified cells, less than or about 10%, less than or about 9%, less than or about 8%, less than or about 7%, less than or about 7%, less than or about 5%, less than or about 4%, less than or about 4%, less than or about 3%, less than or about 2%, or less than or about 1% of the cells of said plurality comprise a chimeric antigen receptor that exhibits tonic signaling and / or antigen-independent activity or signaling.

[0097] Also provided are compositions containing any of the chimeric receptors provided herein. Also provided are compositions containing any of the modified cells provided herein. In some of any of the provided embodiments, the compositions comprise CD4+ T cells and CD8+ T cells, wherein the ratio of CD4+ T cells to CD8+ T cells is 1:3 to 3:1, or about 1:3 to about 3:1, optionally 1:2 to 2:1, or about 1:2 to about 2:1. In some of any of the provided embodiments, the compositions comprise CD4+ T cells and CD8+ T cells, wherein the ratio of CD4+ T cells to CD8+ T cells is about 1:1.

[0098] Also provided are compositions containing a plurality of first modified cells comprising a first chimeric antigen receptor that is any of the chimeric antigen receptors provided herein or encoded by any of the polynucleotides provided herein; and a plurality of second modified cells comprising a second chimeric antigen receptor. In some of any of the provided embodiments, of the plurality of first modified cells, less than or about 10%, less than or about 9%, less than or about 8%, less than or about 7%, less than or about 5%, less than or about 4%, less than or about 3%, less than or about 2%, or less than or about 1% of the cells of the plurality comprise a chimeric antigen receptor that exhibits tonic signaling and / or antigen-independent activity or signaling. In some of any of the provided embodiments, of a plurality of said second modified cells, less than or about 10%, less than or about 9%, less than or about 8%, less than or about 7%, less than or about 5%, less than or about 4%, less than or about 3%, less than or about 2%, or less than or about 1% of the cells of said plurality comprise a chimeric antigen receptor that exhibits tonic signaling and / or antigen-independent activity or signaling.

[0099] In some of any of the provided embodiments, the second chimeric receptor of the plurality of second modified cells in the composition comprises an extracellular antigen-binding domain that specifically binds to a second antigen expressed on or associated with multiple myeloma. In some of any of the provided embodiments, the second CAR of the plurality of second modified cells in the composition comprises an extracellular antigen-binding domain that binds to the second antigen, a spacer, a transmembrane domain, and an intracellular signaling region. In some of any of the provided embodiments, the second antigen of the plurality of second modified cells in the composition is selected from B-cell maturation antigen (BCMA), CD38, CD138, CS-1, BAFF-R, TACI, or FcRH5. In some of any of the provided embodiments, the second antigen of the plurality of second modified cells in the composition is BCMA.

[0100] In some of any of the provided embodiments, the second CAR of the plurality of second modified cells in the composition comprises (1) an extracellular antigen-binding domain that specifically binds to BCMA, the extracellular antigen-binding domain being (i) a V as set forth in any of SEQ ID NOs: 189, 191, 193, 195, or 197. H a heavy chain variable region (V) comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the V region amino acid sequence; H and (ii) V as set forth in any of SEQ ID NOs: 190, 192, 194, 196, or 198. L a light chain variable (V) region comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the V region amino acid sequence; L (1) an extracellular antigen-binding domain comprising a V region; (2) a spacer; (3) a transmembrane region; and (4) an intracellular signaling region.H The region may be any of the V sequences shown in SEQ ID NOs: 189, 191, 193, 195 or 197. H V includes CDR-H1, CDR-H2, and CDR-H3 contained within the amino acid sequence of the region; L The region may be any of the V sequences shown in SEQ ID NOs: 190, 192, 194, 196, or 198. L The regions include CDR-L1, CDR-L2, and CDR-L3 contained within the amino acid sequence.

[0101] In some of any of the provided embodiments, the second CAR of the plurality of second modified cells in the composition comprises (1) a heavy chain variable region (V) that specifically binds to BCMA, the V V V heavy chain variable region (V) comprising: (i) a heavy chain complementarity determining region 1 (CDR-H1) comprising an amino acid sequence selected from any one of SEQ ID NOs: 199, 202, 206, 209, 212, or 215; (b) a heavy chain complementarity determining region 2 (CDR-H2) comprising an amino acid sequence selected from any one of SEQ ID NOs: 200, 203, 207, 210, 213, or 216; and (c) a heavy chain complementarity determining region 3 (CDR-H3) comprising an amino acid sequence selected from any one of SEQ ID NOs: 201, 204, 205, 208, 211, 214, or 217. H and (ii) a light chain complementarity determining region 1 (CDR-L1) comprising an amino acid sequence selected from any one of SEQ ID NOs: 218, 221, 224, 227, 230, 233, or 235; (b) a light chain complementarity determining region 2 (CDR-L2) comprising an amino acid sequence selected from any one of SEQ ID NOs: 219, 222, 225, 228, 231, 234, or 236; and (c) a light chain complementarity determining region 3 (CDR-L3) comprising an amino acid sequence selected from any one of SEQ ID NOs: 220, 223, 226, 229, or 232. L (1) an extracellular antigen-binding domain, including a spacer region; (2) a transmembrane domain; and (3) an intracellular signaling region.

[0102] In some of any of the provided embodiments, the V of the second CAR of the composition H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 199, 200, and 201, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 218, 219, and 220, respectively; H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 202, 203, and 204, respectively, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 221, 222, and 223, respectively; H The regions include CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 199, 200, and 205, respectively, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 224, 225, and 226, respectively; H The regions include CDR-H1, CDR-H2, and CDR-H3 containing the amino acid sequences of SEQ ID NOs: 206, 207, and 208, respectively, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 227, 228, and 229, respectively; H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; HThe regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 212, 213, and 214, respectively, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 233, 234, and 229, respectively; or the V of the second CAR H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 215, 216, and 217, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3, which comprise the amino acid sequences of SEQ ID NOs: 235, 236, and 232, respectively.

[0103] In some of any of the provided embodiments, the V of the second CAR in the composition H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; or the V of the second CAR H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 215, 216, and 217, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3, which comprise the amino acid sequences of SEQ ID NOs: 235, 236, and 232, respectively.

[0104] In some of any of the provided embodiments, the V of the second CAR in the composition H Area and V LThe regions comprise the amino acid sequences set forth in SEQ ID NO: 189 and SEQ ID NO: 190, respectively, or an amino acid sequence that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 189 and SEQ ID NO: 190; H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NO: 191 and SEQ ID NO: 192, respectively, or an amino acid sequence that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 191 and SEQ ID NO: 192; H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NO: 193 and SEQ ID NO: 194, respectively, or an amino acid sequence that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 193 and SEQ ID NO: 194; H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NO: 195 and SEQ ID NO: 196, respectively, or an amino acid sequence that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 195 and SEQ ID NO: 196; or H Area and V LThe regions comprise the amino acid sequences set forth in SEQ ID NO:197 and SEQ ID NO:198, respectively, or amino acid sequences that exhibit at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:197 and SEQ ID NO:198.

[0105] In some of any of the provided embodiments, the V of the second CAR in the composition H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NO: 189 and SEQ ID NO: 190, respectively; or the V of the second CAR in the composition H Area and V L The region comprises the amino acid sequence shown in SEQ ID NO: 191 and SEQ ID NO: 192; or the V of the second CAR in the composition H Area and V L The region comprises the amino acid sequence shown in SEQ ID NO: 193 and SEQ ID NO: 194; or the V of the second CAR in the composition H Area and V L The region comprises the amino acid sequence shown in SEQ ID NO: 195 and SEQ ID NO: 196; or the V of the second CAR in the composition H Area and V L The regions comprise the amino acid sequences shown in SEQ ID NO:197 and SEQ ID NO:198, respectively.

[0106] In some of any of the provided embodiments, the extracellular antigen-binding domain of the second CAR in the composition is a single-chain antibody fragment. In some such embodiments, the fragment is or comprises a single-chain variable fragment (scFv). In some of any of the provided embodiments, the V of the second CAR in the composition H Area and V LThe regions are connected by a flexible linker. In some of any of the provided embodiments, the linker in the second CAR in the composition has the amino acid sequence In some of any of the provided embodiments, the linker in the second CAR in the composition has the amino acid sequence Includes TIFF2025183309000017.tif4128.

[0107] In some of any of the provided embodiments, V is present in a second CAR in the composition. H The area is V L In some of the provided embodiments, a V H The area is V L In some of any of the provided embodiments, the antigen-binding domain in the second CAR in the composition comprises an amino acid sequence selected from any one of SEQ ID NOs: 227, 238, 239, 240, or 241, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence selected from any one of SEQ ID NOs: 227, 238, 239, 240, or 241. In some of any of the provided embodiments, the antigen-binding domain in the second CAR in the composition comprises an amino acid sequence selected from any one of SEQ ID NOs: 227, 238, 239, 240, or 241.

[0108] In some of any of the provided embodiments, the V of the second CAR in the composition H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and the V of the second CARL The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; or the V of the second CAR H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 215, 216, and 217, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 235, 236, and 232, respectively; and / or the V of the second CAR in the composition. H and / or the antigen-binding domain of a second CAR in the composition comprises the amino acid sequence set forth in SEQ ID NO:241, or a sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:241. In some of any of the provided embodiments, the VL region and VL region of a second CAR in the composition comprise the amino acid sequence set forth in SEQ ID NO:197 and SEQ ID NO:198, respectively. H The regions include CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and the V of the second CAR L The regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; and / or the V of the second CAR in the composition. Hthe VL region and VL region comprise the amino acid sequences set forth in SEQ ID NO:197 and SEQ ID NO:198, respectively; and / or the antigen-binding domain of the second CAR in the composition comprises the amino acid sequence set forth in SEQ ID NO:241, or a sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:241.

[0109] In some of any of the provided embodiments, the transmembrane domain of the second CAR in the composition is or comprises a transmembrane domain derived from CD4, CD28, or CD8, optionally a transmembrane domain derived from human CD4, human CD28, or human CD8. and / or the transmembrane domain of the second CAR in the composition is or comprises the sequence set forth in SEQ ID NO: 18, or an amino acid sequence that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 18. In some of any of the provided embodiments, the transmembrane domain of the second CAR in the composition is or comprises the sequence set forth in SEQ ID NO: 18.

[0110] In some of any of the provided embodiments, the intracellular signaling region of the second CAR in the composition comprises an intracellular signaling domain. In some of any of the provided embodiments, the intracellular signaling domain of the second CAR in the composition is capable of inducing a primary activation signal in a T cell, is a component of a T cell receptor (TCR), and / or comprises an immunoreceptor tyrosine-based activation motif (ITAM). In some of any of the provided embodiments, the intracellular signaling domain of the second CAR in the composition is or comprises the cytoplasmic signaling domain of the zeta chain of the CD3-zeta (CD3ζ) chain or a functional variant or signaling portion thereof, optionally the cytoplasmic signaling domain of the human CD3 zeta chain. In some of any of the provided embodiments, the intracellular signaling region of the second CAR in the composition comprises the sequence set forth in SEQ ID NO:20, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:20.

[0111] In some of any of the provided embodiments, the intracellular signaling region of the second CAR in the composition further comprises a costimulatory signaling region. In some of any of the provided embodiments, the costimulatory signaling region of the second CAR in the composition comprises the intracellular signaling domain of a T cell costimulatory molecule or a signaling portion thereof. In some of any of the provided embodiments, the costimulatory signaling region of the second CAR in the composition comprises the intracellular signaling domain of CD28, 4-1BB, or ICOS, or a signaling portion thereof, optionally the intracellular signaling domain of human CD28, human 4-1BB, or human ICOS. In some of any of the provided embodiments, the costimulatory signaling region of the second CAR in the composition comprises the intracellular signaling domain of 4-1BB or a signaling portion thereof, optionally the intracellular signaling domain of human 4-1BB. In some of any of the provided embodiments, at least one of the first chimeric antigen receptor and the second chimeric antigen receptor comprises an intracellular signaling region comprising the intracellular signaling domain of 4-1BB or a signaling portion thereof, optionally the intracellular signaling domain of human 4-1BB. In some of any of the provided embodiments, the costimulatory signaling region of the second CAR in the composition comprises the intracellular signaling domain of human CD28; and / or the sequence set forth in SEQ ID NO:46, or an amino acid sequence exhibiting at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:46.In some of any of the provided embodiments, the costimulatory signaling region of the second CAR in the composition comprises the intracellular signaling domain of human 4-1BB; and / or the sequence set forth in SEQ ID NO:19, or an amino acid sequence that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:19.

[0112] In some of any of the provided embodiments, the encoded second chimeric antigen receptor of the composition comprises, in N- to C-terminal order, an antigen-binding domain, a spacer, a transmembrane domain, and an intracellular signaling region.

[0113] In some of any of the provided embodiments, the plurality of first modified cells in the composition comprises T cells, optionally comprising CD4+ T cells and CD8+ T cells, and optionally a ratio of CD4+ T cells to CD8+ T cells of 1:3 to 3:1, or about 1:3 to about 3:1, or optionally 1:2 to 2:1, or about 1:2 to about 2:1. In some of any of the provided embodiments, the plurality of first modified cells in the composition comprises T cells, optionally comprising CD4+ T cells and CD8+ T cells, and optionally a ratio of CD4+ T cells to CD8+ T cells of about 1:1.

[0114] In some of any of the provided embodiments, the plurality of second modified cells in the composition comprises T cells, optionally comprising CD4+ T cells and CD8+ T cells, and optionally a ratio of CD4+ T cells to CD8+ T cells of 1:3 to 3:1, or about 1:3 to about 3:1, or optionally 1:2 to 2:1, or about 1:2 to about 2:1. In some of any of the provided embodiments, the plurality of second modified cells in the composition comprises T cells, optionally comprising CD4+ T cells and CD8+ T cells, and optionally a ratio of CD4+ T cells to CD8+ T cells of about 1:1.

[0115] In some of any of the provided embodiments, the composition comprises a ratio of a first plurality of modified cells to a second plurality of modified cells of 1:3 to 3:1 or about 1:3 to 3:1, optionally 1:2 to 2:1 or about 1:2 to 2:1, and optionally 1:1 or about 1:1. In some of any of the provided embodiments, the composition comprises a first plurality of cells expressing a first chimeric antigen receptor to a second plurality of cells expressing a second chimeric antigen receptor in a ratio of about 1:3 to 3:1, optionally about 1:2 to 2:1. In certain embodiments, the ratio of the first plurality of modified cells to the second plurality of modified cells in the composition is 1:1 or about 1:1. In some of any of the provided embodiments, the composition further comprises a pharmaceutically acceptable excipient. In some of any of the provided embodiments, the composition is sterile.

[0116] Also provided herein are uses of any of the compositions provided herein. In some of any of the provided embodiments, the compositions are for use in treating a subject with a disease or condition. In some of any of the provided embodiments, the disease or condition is cancer. In some of any of the provided embodiments, the disease or condition is multiple myeloma, optionally relapsed / refractory multiple myeloma. The provided uses and compositions for use provided herein can be for treating a subject according to any aspect of the provided methods.

[0117] Also provided herein are methods of treatment that include administering to a subject having a disease or disorder any of the compositions provided herein comprising any of the modified cells provided herein or any of the compositions provided herein comprising any of the chimeric antigen receptors provided herein. In some of any of the provided embodiments, the dose of cells is 1.0 x 10 7 or approximately 1.0 x 10 7 CAR-expressing T cells ~1.2 x 109 or approximately 1.2 x 10 9 CAR-expressing T cells, approximately 1.25 x 10 7 CAR-expressing T cells ~ approximately 1.2 x 10 9 CAR-expressing T cells, approximately 1.5 x 10 7 CAR-expressing T cells ~ approximately 1.2 x 10 9 CAR-expressing T cells, approximately 5.0 x 10 7 CAR-expressing T cells ~ approximately 4.5 x 10 8 CAR-expressing T cells, approximately 1.5 x 10 8 CAR-expressing T cells ~approximately 3.0 x 10 8 In some of any of the provided embodiments, the dose of cells comprises 2.5 x 10 CAR-expressing T cells. 7 or approximately 2.5 x 10 7 CAR-expressing T cells ~1.2 x 10 9 or approximately 1.2 x 10 9 CAR-expressing T cells, 5.0 x 10 7 or approximately 5.0 x 10 7 CAR-expressing T cells ~4.5 x 10 8 or approximately 4.5 x 10 8 CAR-expressing T cells, 1.5 x 10 8 or about 1.5 x 10 8 CAR-expressing T cells ~3.0 x 10 8 or approximately 3.0 x 10 8 Contains CAR-expressing T cells.

[0118] In some of either embodiment, the dose of cells is 1×10 7 Or about 1 x 10 7 CAR-expressing T cells ~2 x 10 9 Or about 2 x 10 9 In some of either embodiment, the dose of cells comprises 2.5 x 10 CAR-expressing T cells. 7 Or about 2.5 x 10 7 CAR-expressing T cells ~1.2 x 10 9 or about 1.2 x 10 9 CAR-expressing T cells, 5.0 x 10 7 or approximately 5.0 x 10 7CAR-expressing T cells ~4.5 x 10 8 Or about 4.5 x 10 8 CAR-expressing T cells, or 1.5 x 10 8 Or about 1.5 x 10 8 CAR-expressing T cells ~3.0 x 10 8 Or about 3.0 x 10 8 In some of either embodiment, the dose of cells comprises 1.0 x 10 CAR-expressing T cells. 7 or approximately 1.0 x 10 7 pieces, 1.5×10 7 Or about 1.5 x 10 7 pieces, 2.5×10 7 Or about 2.5 x 10 7 pieces, 5.0×10 7 or approximately 5.0 x 10 7 pieces, 7.5×10 7 Or about 7.5 x 10 7 pieces, 1.5×10 8 Or about 1.5 x 10 8 pieces, 2.25×10 8 Or about 2.25 x 10 8 pieces, 3.0×10 8 Or about 3.0 x 10 8 pieces, 4.5×10 8 or about 4.5 x 10 8 pieces, 6.0×10 8 Or about 6.0 x 10 8 pieces, 8.0×10 8 or approximately 8.0 x 10 8 pieces, or 1.2 x 10 9 or about 1.2 x 10 9 In some of either embodiment, the dose of cells comprises 5.0 x 10 CAR-expressing T cells. 7 or approximately 5.0 x 10 7 pieces, 1.5×10 8 Or about 1.5 x 10 8 pieces, 3.0×10 8 Or about 3.0 x 10 8 pieces, or 4.5 x 10 8 or about 4.5 x 10 8In some of either embodiment, the dose of cells comprises 5.0 x 10 CAR-expressing T cells. 7 or approximately 5.0 x 10 7 pieces, 1.5×10 8 Or about 1.5 x 10 8 pieces, 3.0×10 8 Or about 3.0 x 10 8 pieces, or 4.5 x 10 8 or about 4.5 x 10 8 In some of either embodiment, the dose of cells comprises 5.0 x 10 CAR-expressing T cells. 7 or approximately 5.0 x 10 7 Contains CAR-expressing T cells.

[0119] Also provided herein are uses of a first composition containing a plurality of first modified cells comprising a first chimeric antigen receptor, the first chimeric antigen receptor being any of the chimeric antigen receptors provided herein or any chimeric antigen receptor encoded by any of the polynucleotides provided herein, and a second composition containing a plurality of second modified cells comprising a second chimeric antigen receptor. In some of any of the provided embodiments, the compositions are used together for use in treating a subject with a disease or condition. In some of any of the provided embodiments, the disease or condition is cancer. In some of any of the provided embodiments, the disease or condition is multiple myeloma, optionally relapsed / refractory multiple myeloma. The provided uses and compositions for use provided herein may be for treating a subject according to any of the provided method aspects.

[0120] Also provided herein are methods of treatment comprising administering to a subject having a disease or disorder a composition containing a plurality of first modified cells comprising a first chimeric antigen receptor, the first chimeric antigen receptor being any of the chimeric antigen receptors provided herein or any chimeric antigen receptor encoded by any of the polynucleotides provided herein; and administering to the subject a composition containing a plurality of second modified cells comprising a second chimeric antigen receptor. In some of any of the provided embodiments, the dose of the first plurality of modified cells and the dose of the second plurality of modified cells are independently 1.0 x 10 7 or approximately 1.0 x 10 7 CAR-expressing T cells ~1.5 x 10 9 Or about 1.5 x 10 9 CAR-expressing T cells, 1.25 x 10 7 Or about 1.25 x 10 7 CAR-expressing T cells ~0.6 x 10 8 or approximately 0.6 x 10 8 CAR-expressing T cells, 2.5 x 10 7 Or about 2.5 x 10 7 CAR-expressing T cells ~2.25 x 10 8 Or about 2.25 x 10 8 CAR-expressing T cells, 7.5 x 10 7 Or about 7.5 x 10 7 CAR-expressing T cells ~1.5 x 10 8 Or about 1.5 x 10 8 CAR-expressing T cells, 2.5 x 10 7 Or about 2.5 x 10 7 CAR-expressing T cells ~1.2 x 10 9 or about 1.2 x 10 9 CAR-expressing T cells, 5.0 x 10 7 or approximately 5.0 x 10 7 CAR-expressing T cells ~4.5 x 10 8 or about 4.5 x 10 8 CAR-expressing T cells, 1.5 x 10 8 Or about 1.5 x 10 8 CAR-expressing T cells ~3.0 x 10 8Or about 3.0 x 10 8 In some of any of the embodiments, the dose of the first plurality of modified cells and the dose of the second plurality of modified cells are independently 1 x 10 or more than 1 x 10 7 Or about 1 x 10 7 CAR-expressing T cells ~2 x 10 9 Or about 2 x 10 9 In some of either embodiment, the dose of cells comprises 2.5 x 10 CAR-expressing T cells. 7 Or about 2.5 x 10 7 CAR-expressing T cells ~1.2 x 10 9 or about 1.2 x 10 9 CAR-expressing T cells, 5.0 x 10 7 or approximately 5.0 x 10 7 CAR-expressing T cells ~4.5 x 10 8 or about 4.5 x 10 8 CAR-expressing T cells, or 1.5 x 10 8 Or about 1.5 x 10 8 CAR-expressing T cells ~3.0 x 10 8 Or about 3.0 x 10 8 In some of either embodiment, the dose of cells comprises 1.5 x 10 CAR-expressing T cells. 7 Or about 1.5 x 10 7 pieces, 2.5×10 7 Or about 2.5 x 10 7 pieces, 5.0×10 7 or approximately 5.0 x 10 7 pieces, 7.5×10 7 Or about 7.5 x 10 7 pieces, 1.5×10 8 Or about 1.5 x 10 8 pieces, 2.25×10 8 Or about 2.25 x 10 8 pieces, 3.0×10 8 Or about 3.0 x 10 8 pieces, 4.5×10 8 or about 4.5 x 10 8 pieces, 6.0×10 8 Or about 6.0 x 10 8 pieces, 8.0×10 8or approximately 8.0 x 10 8 pieces, or 1.2 x 10 9 or about 1.2 x 10 9 In some of either embodiment, the dose of cells comprises 5.0 x 10 CAR-expressing T cells. 7 or approximately 5.0 x 10 7 pieces, 1.5×10 8 Or about 1.5 x 10 8 pieces, 3.0×10 8 Or about 3.0 x 10 8 or 4.5 x 10 8 or about 4.5 x 10 8 In some of either embodiment, the dose of cells comprises 5.0 x 10 CAR-expressing T cells. 7 or approximately 5.0 x 10 7 pieces, 1.5×10 8 Or about 1.5 x 10 8 pieces, 3.0×10 8 Or about 3.0 x 10 8 pieces, or 4.5 x 10 8 or about 4.5 x 10 8 In some of any of the embodiments, the dose of cells comprises 5.0 x 10 CAR-expressing T cells. 7 or approximately 5.0 x 10 7 Contains CAR-expressing T cells.

[0121] In some of any of the provided embodiments, the composition containing the first plurality of modified cells and the composition containing the second plurality of modified cells are administered simultaneously, sequentially, or intermittently. In some of any of the provided embodiments, the composition containing the first plurality of modified cells and the composition containing the second plurality of modified cells are administered sequentially in any order.

[0122] In some of any of the provided embodiments, of a plurality of said first modified cells of a composition in a provided method of treatment or a composition for use in a treatment, less than or about 10%, less than or about 9%, less than or about 8%, less than or about 7%, less than or about 5%, less than or about 4%, less than or about 3%, less than or about 2%, or less than or about 1% of the cells of said plurality comprise a chimeric antigen receptor that exhibits tonic signaling and / or antigen-independent activity or signaling.

[0123] In some of any of the provided embodiments, of a plurality of said second modified cells of a composition in a provided method of treatment or a composition for use in a treatment, less than or about 10%, less than or about 9%, less than or about 8%, less than or about 7%, less than or about 5%, less than or about 4%, less than or about 3%, less than or about 2%, or less than or about 1% of the cells of said plurality comprise a chimeric antigen receptor that exhibits tonic signaling and / or antigen-independent activity or signaling.

[0124] In some of any of the provided embodiments, the second chimeric receptor of the composition in the method of treatment or the modified cell of the composition for use in treatment comprises an extracellular antigen-binding domain that specifically binds to a second antigen expressed on or associated with multiple myeloma.

[0125] In some of any of the provided embodiments, the second CAR of the modified cells of the provided methods of treatment or compositions for use in treatment comprises an extracellular antigen-binding domain that binds to a second antigen, a spacer, a transmembrane domain, and an intracellular signaling region.

[0126] In some of any of the provided embodiments, the second antigen targeted by the second CAR of the modified cells of the provided methods of treatment or compositions for use in treatment includes or is selected from B-cell maturation antigen (BCMA), CD38, CD138, CS-1, BAFF-R, TACI, or FcRH5. In some of any of the provided embodiments, the second antigen in the provided methods includes or is BCMA.

[0127] In some of any of the provided embodiments, the second CAR of the engineered cells of the provided methods of treatment or compositions for use in treatment comprises (1) an extracellular antigen-binding domain that specifically binds to BCMA, the extracellular antigen-binding domain being (i) a V as set forth in any of SEQ ID NOs: 189, 191, 193, 195, or 197. H a heavy chain variable region (V) comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the V region amino acid sequence; H and (ii) V as set forth in any of SEQ ID NOs: 190, 192, 194, 196, or 198. L a light chain variable (V) region comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the V region amino acid sequence; L (1) an extracellular antigen-binding domain, including a V region; (2) a spacer; (3) a transmembrane region; and (4) an intracellular signaling region. In some of any of the provided embodiments, the V region of the second CAR in the provided methods comprises H The region may be any of the V sequences shown in SEQ ID NOs: 189, 191, 193, 195 or 197. H V includes CDR-H1, CDR-H2, and CDR-H3 contained within the amino acid sequence of the region; LThe region may be any of the V sequences shown in SEQ ID NOs: 190, 192, 194, 196, or 198. L The regions include CDR-L1, CDR-L2, and CDR-L3 contained within the amino acid sequence.

[0128] In some of any of the provided embodiments, the second CAR of the engineered cells of the provided methods of treatment or compositions for use in treatment comprises (1) an extracellular antigen-binding domain that specifically binds to BCMA, wherein (i) V H a heavy chain variable region (V) comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:197; H ); and (ii) V L a light chain variable (V) region comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:198; L (1) an extracellular antigen-binding domain, including a V region; (2) a spacer; (3) a transmembrane region; and (4) an intracellular signaling region. In some of any of the provided embodiments, the V region of the second CAR in the provided methods comprises H The region is V as shown in SEQ ID NO:197. H V includes CDR-H1, CDR-H2, and CDR-H3 contained within the amino acid sequence of the region; L The region is V shown in SEQ ID NO:198. L The regions include CDR-L1, CDR-L2, and CDR-L3 contained within the amino acid sequence.

[0129] In some of any of the provided embodiments, the second CAR of the engineered cells of the provided methods of treatment or compositions for use in treatment comprises (1) a heavy chain variable region (V) that specifically binds to BCMA, the VV comprising: (i) a heavy chain complementarity determining region 1 (CDR-H1) comprising an amino acid sequence selected from any one of SEQ ID NOs: 199, 202, 206, 209, 212, or 215; (b) a heavy chain complementarity determining region 2 (CDR-H2) comprising an amino acid sequence selected from any one of SEQ ID NOs: 200, 203, 207, 210, 213, or 216; and (c) a heavy chain complementarity determining region 3 (CDR-H3) comprising an amino acid sequence selected from any one of SEQ ID NOs: 201, 204, 205, 208, 211, 214, or 217. H and (ii) a light chain complementarity determining region 1 (CDR-L1) comprising an amino acid sequence selected from any one of SEQ ID NOs: 218, 221, 224, 227, 230, 233, or 235; (b) a light chain complementarity determining region 2 (CDR-L2) comprising an amino acid sequence selected from any one of SEQ ID NOs: 219, 222, 225, 228, 231, 234, or 236; and (c) a light chain complementarity determining region 3 (CDR-L3) comprising an amino acid sequence selected from any one of SEQ ID NOs: 220, 223, 226, 229, or 232. L (1) an extracellular antigen-binding domain, including a spacer region; (2) a transmembrane domain; and (3) an intracellular signaling region.

[0130] In some of any of the provided embodiments, the second CAR of the modified cells of the provided methods of treatment or compositions for use in treatment comprises the V of the second CAR. H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 199, 200, and 201, respectively; Lthe regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 218, 219, and 220, respectively; H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 202, 203, and 204, respectively; L the regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 221, 222, and 223, respectively; H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 199, 200, and 205, respectively; L the regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 224, 225, and 226, respectively; H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 206, 207, and 208, respectively; L the regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 227, 228, and 229, respectively; H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively; L the regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 212, 213, and 214, respectively; L the regions comprise CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 233, 234, and 229, respectively; or the V of the second CAR HThe second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 215, 216, and 217, respectively; L The region comprises an extracellular antigen-binding domain comprising CDR-L1, CDR-L2, and CDR-L3, which comprise the amino acid sequences of SEQ ID NOs: 235, 236, and 232, respectively.

[0131] In some of any of the provided embodiments, the second CAR of the modified cells of the provided methods of treatment or compositions for use in treatment comprises the V of the second CAR. H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively; L the CDR-L1, CDR-L2, and CDR-L3 regions comprise the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; or the V region of the second CAR in the provided method. H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 215, 216, and 217, respectively; L In some of the provided embodiments, the second CAR of the modified cells of the provided methods of treatment or compositions for use in treatment comprises an extracellular antigen-binding domain comprising CDR-L1, CDR-L2, and CDR-L3, the CDR-L1 region comprising the amino acid sequences of SEQ ID NOs: 235, 236, and 232, respectively. H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively; L The region comprises an extracellular antigen-binding domain comprising CDR-L1, CDR-L2, and CDR-L3, which comprise the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively.

[0132] In some of any of the provided embodiments, the second CAR of the modified cells of the provided methods of treatment or compositions for use in treatment is V H the VL region and VL region of the second CAR comprise the amino acid sequences set forth in SEQ ID NO: 189 and SEQ ID NO: 190, respectively, or amino acid sequences that exhibit at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 189 and SEQ ID NO: 190; H the VL region and VL region of the second CAR comprise the amino acid sequences set forth in SEQ ID NO: 191 and SEQ ID NO: 192, respectively, or amino acid sequences that exhibit at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 191 and SEQ ID NO: 192; H the VL region and VL region of the second CAR comprise the amino acid sequences set forth in SEQ ID NO: 193 and SEQ ID NO: 194, respectively, or amino acid sequences that exhibit at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 193 and SEQ ID NO: 194; H the VL region and VL region of the second CAR comprise the amino acid sequences set forth in SEQ ID NO: 195 and SEQ ID NO: 196, respectively, or an amino acid sequence that exhibits at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 195 and SEQ ID NO: 196; or HThe VL and VL regions comprise an extracellular antigen-binding domain comprising the amino acid sequences set forth in SEQ ID NO:197 and SEQ ID NO:198, respectively, or amino acid sequences exhibiting at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:197 and SEQ ID NO:198.

[0133] In some of any of the provided embodiments, the second CAR of the modified cells of the provided methods of treatment or compositions for use in treatment is V H The VL and VL regions comprise an extracellular antigen-binding domain comprising the amino acid sequences set forth in SEQ ID NO:197 and SEQ ID NO:198, respectively, or amino acid sequences exhibiting at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:197 and SEQ ID NO:198.

[0134] In some of any of the provided embodiments, the second CAR of the modified cells of the provided methods of treatment or compositions for use in treatment is V H the VL region and VL region of the second CAR comprise the amino acid sequences set forth in SEQ ID NO: 189 and SEQ ID NO: 190, respectively; H the VL region of the second CAR comprises the amino acid sequences set forth in SEQ ID NO: 191 and SEQ ID NO: 192; H the VL region of the second CAR comprises the amino acid sequences set forth in SEQ ID NO: 193 and SEQ ID NO: 194; H the VL region and VL region of the second CAR comprise the amino acid sequences set forth in SEQ ID NO: 195 and SEQ ID NO: 196; or HIn some of the provided embodiments, the second CAR of the modified cells of the provided methods of treatment or compositions for use in treatment comprises an extracellular antigen-binding domain in which the VL region and the VL region comprise the amino acid sequences set forth in SEQ ID NO: 197 and SEQ ID NO: 198, respectively. H The VL and VL regions comprise an extracellular antigen-binding domain comprising the amino acid sequences set forth in SEQ ID NO:197 and SEQ ID NO:198, respectively.

[0135] In some of any of the provided embodiments, the extracellular antigen-binding domain of the second CAR of the engineered cell in the provided methods of treatment or compositions for use in treatment is a single-chain antibody fragment. In some of any of the provided embodiments, the fragment is or comprises a single-chain variable fragment (scFv).

[0136] In some of the provided embodiments of the provided methods or some of the provided embodiments for use in therapy, the V of the extracellular antigen-binding domain of the second CAR of the modified cell in the composition H Area and V L The regions are connected by a flexible linker. In some of any of the provided embodiments, the linker of the second CAR of the engineered cell in the composition has the amino acid sequence In some of any of the provided embodiments, the linker of the second CAR of the engineered cell in the composition has the amino acid sequence Includes TIFF2025183309000019.tif4128.

[0137] In some of the provided embodiments of the provided methods or some of the provided embodiments for use in therapy, a V H The area is V LIn some of any of the provided embodiments of the provided methods, a V H The area is V L It is carboxy-terminal to the region.

[0138] In some of the provided embodiments of the provided methods, or some of the provided embodiments for use in therapy, the extracellular antigen-binding domain of the second CAR comprises an amino acid sequence selected from any one of SEQ ID NOs: 227, 238, 239, 240, or 241, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence selected from any one of SEQ ID NOs: 227, 238, 239, 240, or 241. In some of the provided embodiments of the provided methods, the antigen-binding domain of the second CAR comprises an amino acid sequence selected from any one of SEQ ID NOs: 227, 238, 239, 240, or 241. In some of the provided embodiments of the provided methods or some of the provided embodiments for use in therapy, the extracellular antigen-binding domain of the second CAR comprises the amino acid sequence set forth in SEQ ID NO:241, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:241. In some of the provided embodiments of the provided methods, the antigen-binding domain of the second CAR comprises the amino acid sequence set forth in SEQ ID NO:241.

[0139] In some of the provided embodiments of the provided methods or some of the provided embodiments for use in therapy, the extracellular antigen-binding domain of the second CAR is H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively; L the regions comprise CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; or the V of the second CAR H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 215, 216, and 217, respectively; L the regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 235, 236, and 232, respectively; and / or the V of the second CAR H The VL and VL regions comprise the amino acid sequences set forth in SEQ ID NO: 197 and SEQ ID NO: 198, respectively. H Area and V L and / or the antigen-binding domain of the second CAR comprises the amino acid sequence set forth in SEQ ID NO:241, or a sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:241. In some of the provided embodiments of the provided methods or some of the provided embodiments for use in therapy, the extracellular antigen-binding domain of the second CAR comprises the V H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively; Lthe regions include CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; and / or the V of the second CAR H The VL and VL regions comprise the amino acid sequences set forth in SEQ ID NO: 197 and SEQ ID NO: 198, respectively. H Area and V L and / or the antigen-binding domain of the second CAR comprises the amino acid sequence set forth in SEQ ID NO:241, or a sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:241.

[0140] In some of any of the provided embodiments of the provided methods or some of the provided embodiments for use in therapy, the transmembrane domain of the second CAR is or comprises a transmembrane domain derived from CD4, CD28, or CD8, optionally a transmembrane domain derived from human CD4, human CD28, or human CD8. In some of the provided embodiments of the provided methods, the transmembrane domain of the second CAR is or comprises a transmembrane domain derived from human CD28; and / or the transmembrane domain is or comprises the sequence set forth in SEQ ID NO: 18, or an amino acid sequence exhibiting at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 18. In some of the provided embodiments of the provided methods, the transmembrane domain of the second CAR is or comprises the sequence set forth in SEQ ID NO: 18.

[0141] In some of the provided embodiments of the provided methods or some of the provided embodiments for use in therapy, the intracellular signaling region of the second CAR comprises an intracellular signaling domain. In some of the provided embodiments of the provided methods, the intracellular signaling domain of the second CAR is capable of inducing a primary activation signal in a T cell, is a component of a T cell receptor (TCR), and / or comprises an immunoreceptor tyrosine-based activation motif (ITAM). In some of the provided embodiments of the provided methods, the intracellular signaling domain of the second CAR is or comprises the cytoplasmic signaling domain of the zeta chain of the CD3-zeta (CD3ζ) chain or a functional variant or signaling portion thereof, optionally the cytoplasmic signaling domain of the human CD3 zeta chain. In some of any of the provided embodiments of the provided methods, the intracellular signaling region of the second CAR comprises the sequence set forth in SEQ ID NO:20, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:20.

[0142] In some of the provided embodiments of the provided methods or some of the provided embodiments for use in therapy, the intracellular signaling region of the second CAR further comprises a costimulatory signaling region. In some of the provided embodiments of the provided methods, the costimulatory signaling region of the second CAR comprises the intracellular signaling domain of a T cell costimulatory molecule, or a signaling portion thereof. In some of the provided embodiments of the provided methods, the costimulatory signaling region of the second CAR comprises the intracellular signaling domain of CD28, 4-1BB, or ICOS, or a signaling portion thereof, optionally the intracellular signaling domain of human CD28, human 4-1BB, or human ICOS. In some of the provided embodiments of the provided methods, the costimulatory signaling region of the second CAR comprises the intracellular signaling domain of 4-1BB, or a signaling portion thereof, optionally the intracellular signaling domain of human 4-1BB. In some of any of the provided embodiments, at least one of the first chimeric antigen receptor and the second chimeric antigen receptor comprises an intracellular signaling region that comprises the intracellular signaling domain of 4-1BB or a signaling portion thereof, optionally an intracellular signaling region that comprises the intracellular signaling domain of human 4-1BB.

[0143] In some of the provided embodiments of the provided methods or some of the provided embodiments for use in a therapy, the costimulatory signaling region of the second CAR comprises the intracellular signaling domain of human CD28; and / or the sequence set forth in SEQ ID NO:46, or an amino acid sequence exhibiting at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:46.

[0144] In some of the provided embodiments of the provided methods or some of the provided embodiments for use in therapy, the costimulatory signaling region of the second CAR comprises the intracellular signaling domain of human 4-1BB; and / or the sequence set forth in SEQ ID NO:19, or an amino acid sequence exhibiting at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:19.

[0145] In some of the provided embodiments of the provided methods or some of the provided embodiments for use in therapy, the encoded second chimeric antigen receptor comprises, in order from its N-terminus to C-terminus, an antigen-binding domain, a spacer, a transmembrane domain, and an intracellular signaling region.

[0146] Among the polynucleotides provided herein are polynucleotides comprising: (i) a first nucleic acid sequence encoding a first chimeric antigen receptor (CAR) comprising a first antigen-binding domain; and (ii) a second nucleic acid sequence encoding a second chimeric antigen receptor (CAR) comprising a second antigen-binding domain, wherein the first CAR and the second CAR each comprise: (a) the first antigen-binding domain or the second antigen-binding domain, (b) a spacer, (c) a transmembrane domain, and (d) an intracellular signaling region comprising an intracellular signaling domain and a costimulatory signaling region, wherein one or more of (b) through (d) of the first CAR and the same one or more of (b) through (d) of the second CAR comprise the same amino acid sequence; and the nucleotide sequence encoding one or more of (b) through (d) of the first CAR differs in sequence from the nucleotide sequence encoding the same one or more of (b) through (d) of the second CAR.

[0147] Also disclosed is a polynucleotide comprising: (i) a first nucleic acid sequence encoding a first chimeric antigen receptor (CAR) comprising a first antigen-binding domain capable of binding to one of GPRC5D or BCMA; and (ii) a second nucleic acid sequence encoding a second chimeric antigen receptor (CAR) comprising a second antigen-binding domain capable of binding to the other of GPRC5D or BCMA, wherein the first CAR and the second CAR each comprise: (a) the first antigen-binding domain or the second antigen-binding domain, (b) a spacer, (c) a membrane Also provided herein are polynucleotides comprising (b) a transmembrane domain, and (d) an intracellular signaling region comprising an intracellular signaling domain and a costimulatory signaling region, wherein one or more of (b)-(d) of the first CAR and the same one or more of (b)-(d) of the second CAR comprise identical amino acid sequences; and wherein the nucleotide sequence encoding one or more of (b)-(d) of the first CAR differs in sequence from the nucleotide sequence encoding the same one or more of (b)-(d) of the second CAR.

[0148] In some of any of the provided embodiments, the first antigen-binding domain and the second antigen-binding domain bind to the same antigen. In some of any of the provided embodiments, the first antigen-binding domain and the second antigen-binding domain bind to different epitopes of the same antigen. In some of any of the provided embodiments, the first antigen-binding domain and the second antigen-binding domain bind to different antigens. In some of any of the provided embodiments, the first antigen-binding domain binds to a first antigen expressed by or associated with cells of a disease or pathology, and the second antigen-binding domain binds to a second antigen expressed by or associated with cells of the same disease or pathology.

[0149] In some of any of the provided embodiments, the disease or condition is cancer. In some of any of the provided embodiments, the disease or condition is a cancer that expresses GPRC5D. In some of any of the provided embodiments, the disease or condition is a cancer that expresses BCMA. In some of any of the provided embodiments, the disease or condition is a cancer that expresses BCMA and GPRC5D. In some of any of the provided embodiments, the cancer is a plasma cell malignancy, and the plasma cell malignancy is multiple myeloma (MM) or plasmacytoma. In some of any of the provided embodiments, the cancer is multiple myeloma. In some of any of the provided embodiments, the cancer is relapsed / refractory multiple myeloma.

[0150] In some of any of the provided embodiments, the first antigen-binding domain and the second antigen-binding domain independently bind to an antigen selected from the group consisting of GPRC5D, BCMA, CD38, CD138, CS-1, BAFF-R, TACI, and FcRH5. In some of any of the provided embodiments, the first antigen-binding domain binds to B-cell maturation antigen (BCMA). In some of any of the provided embodiments, the first antigen-binding domain binds to G protein-coupled receptor class C group 5 member D (GPRC5D). In some of any of the provided embodiments, the second antigen-binding domain binds to BCMA. In some of any of the provided embodiments, the second antigen-binding domain binds to GPRC5D.

[0151] In some of any of the provided embodiments, (a) is or comprises a first antigen-binding domain or a second antigen-binding domain, (b) is or comprises a spacer, (c) is or comprises a transmembrane domain, and (d) is or comprises an intracellular signaling region comprising an intracellular signaling domain and a costimulatory signaling region. In some of any of the provided embodiments, one or more of (b)-(d) is one of (b)-(d). In some of any of the provided embodiments, one or more of (b)-(d) is two of (b)-(d). In some of any of the provided embodiments, one or more of (b)-(d) is each of (b)-(d).

[0152] In some of any of the provided embodiments, (a) is or comprises a first antigen-binding domain or a second antigen-binding domain, (b) is or comprises a spacer, (c) is or comprises a transmembrane domain, and (d) is or comprises an intracellular signaling region comprising an intracellular signaling domain and a costimulatory signaling region. In some of any of the provided embodiments, the nucleotide sequence encoding one or more of (a)-(d) of the first CAR and the nucleotide sequence encoding the same one or more of (a)-(d) of the second CAR comprise about 20 or less consecutive base pairs of sequence homology; and / or the first nucleic acid sequence encoding the first CAR and the second nucleic acid sequence encoding the second CAR comprise about 20 or less consecutive base pairs of sequence homology. In some of any of the provided embodiments, the nucleotide sequence encoding one or more of (a)-(d) of the first CAR and the nucleotide sequence encoding the same one or more of (a)-(d) of the second CAR comprise about 5 or less to about 15 contiguous base pairs of sequence homology; and / or the first nucleic acid sequence encoding the first CAR and the second nucleic acid sequence encoding the second CAR comprise about 5 or less and about 15 contiguous base pairs of sequence homology. In some of any of the provided embodiments, the nucleotide sequence encoding one or more of (a)-(d) of the first CAR and the nucleotide sequence encoding the same one or more of (a)-(d) of the second CAR comprise about 10 or less contiguous base pairs of sequence homology; and / or the first nucleic acid sequence encoding the first CAR and the second nucleic acid sequence encoding the second CAR comprise about 10 or less contiguous base pairs of sequence homology.

[0153] In some of any of the provided embodiments, the first nucleic acid encoding the first CAR and the second nucleic acid encoding the second CAR are separated by a nucleotide sequence encoding a multicistronic element, and optionally the multicistronic element is a bicistronic element. In some of any of the provided embodiments, the multicistronic element is an IRES or a ribosomal skipping sequence or a self-cleaving peptide. In some of any of the provided embodiments, the multicistronic element is a ribosomal skipping sequence or a self-cleaving peptide, and the ribosomal skipping sequence or self-cleaving peptide is a T2A, P2A, E2A, or F2A element. In some of any of the provided embodiments, the nucleotide sequence encoding the one or more multicistronic elements is codon divergent. In some of any of the provided embodiments, the nucleotide sequence encoding T2A is codon divergent. In some of any of the provided embodiments, the nucleotide sequence encoding T2A is or comprises the sequence set forth in SEQ ID NO:319.

[0154] In some of any of the provided embodiments, the first nucleic acid sequence encoding the first CAR is codon-optimized for expression in a human cell. In some of any of the provided embodiments, the second nucleic acid sequence encoding the second CAR is codon-optimized for expression in a human cell. In some of any of the provided embodiments, the polynucleotide is codon-optimized for expression in a human cell. In some of any of the provided embodiments, after transcription of the polynucleotide in a human cell (optionally a human T cell), the mRNA, optionally messenger RNA, transcribed from the polynucleotide exhibits at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% RNA homogeneity. In some of any of the provided embodiments, following transcription in a human cell (optionally a human T cell) of a first nucleic acid encoding a first CAR of the polynucleotide, the mRNA, optionally messenger RNA, transcribed from the first nucleic acid exhibits at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% RNA homogeneity. In some of any of the provided embodiments, following transcription in a human cell (optionally a human T cell) of a second nucleic acid encoding a second CAR of the polynucleotide, the mRNA, optionally messenger RNA, transcribed from the second nucleic acid exhibits at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% RNA homogeneity.

[0155] In some of any of the provided embodiments, any potential splice donor sites and / or any potential splice acceptor sites present in the first nucleic acid encoding the first CAR have a nucleotide sequence of less than or at least about 0.70, less than or at least about 0.65, less than or at least about 0.65, less than or at least about 0.60, less than or at least about 0.55, less than or at least about 0.50, less than or at least about 0.45 or at least about 0.40, or less than about 0.35, or at least about 0.30, or less than about 0.25, or at least about 0.20, and / or are predicted to be involved in splice events less than 70%, less than 65%, less than 60%, less than 55%, less than 50%, less than 45%, less than 40%, less than 35%, less than 30%, less than 25%, or less than 20%. In some of any of the provided embodiments, any potential splice donor site or any potential splice acceptor site in the second nucleic acid encoding the second CAR has a RI of less than or at least about 0.70, less than or at least about 0.65, less than or at least about 0.65, less than or at least about 0.60, less than or at least about 0.55, less than or at least about 0.50, less than or at least about 0.45, exhibit a splice junction prediction score of less than or at least about 0.40, less than or at least about 0.35, less than or at least about 0.30, less than or at least about 0.25, less than or at least about 0.20, and / or are predicted to be involved in splice events less than 70%, less than 65%, less than 60%, less than 55%, less than 50%, less than 45%, less than 40%, less than 35%, less than 30%, less than 25%, or less than 20% of the time.In some of any of the provided embodiments, any potential splice donor site or any potential splice acceptor site in the polynucleotide has a β-amino acid sequence of less than about 0.70 or at least less than about 0.70, less than about 0.65 or at least less than about 0.65, less than about 0.60 or at least less than about 0.60, less than about 0.55 or at least less than about 0.55, less than about 0.50 or at least less than about 0.50, less than about 0.45 or at least less than about 0.45, ... or at least about 0.40, or less than about 0.35, or at least about 0.35, or less than about 0.30, or at least about 0.30, or less than about 0.25, or at least about 0.25, or less than about 0.20, and / or are predicted to be involved in splice events less than 70%, less than 65%, less than 60%, less than 55%, less than 50%, less than 45%, less than 40%, less than 35%, less than 30%, less than 25%, or less than 20% of the time.

[0156] In some of any of the provided embodiments, the first antigen-binding domain and / or the second antigen-binding domain of (a) is a single-chain antibody fragment. In some of any of the provided embodiments, the first antigen-binding domain and / or the second antigen-binding domain of (a) is or comprises a single-chain variable fragment (scFv). In some of any of the provided embodiments, the first antigen-binding domain and / or the second antigen-binding domain of (a) comprises a heavy chain variable (VH) region and a light chain variable (VL) region.

[0157] In some of any of the provided embodiments, the first antigen-binding domain or the second antigen-binding domain comprises a VH region comprising CDR-H1 set forth in SEQ ID NO:209, CDR-H2 set forth in SEQ ID NO:210, and CDR-H3 set forth in SEQ ID NO:211, and a VL region comprising CDR-L1 set forth in SEQ ID NO:230, CDR-L2 set forth in SEQ ID NO:231, and CDR-L3 set forth in SEQ ID NO:232. In some of any of the provided embodiments, one of the first antigen-binding domain or the second antigen-binding domain comprises a VH region and a VL region comprising the amino acid sequences set forth in SEQ ID NOs:197 and 198, respectively. In any of the provided embodiments, the first antigen-binding domain or the second antigen-binding domain comprises the amino acid sequence set forth in SEQ ID NO:241, or an amino acid sequence exhibiting at least or at least about 90%, at least or at least about 90%, at least or at least about 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least about 99% sequence identity to SEQ ID NO:241.

[0158] In any of the provided embodiments, one of the first or second antigen-binding domains comprises a VH region comprising CDR-H1 set forth in SEQ ID NO: 125, CDR-H2 set forth in SEQ ID NO: 126, and CDR-H3 set forth in SEQ ID NO: 127, and a VL region comprising CDR-L1 set forth in SEQ ID NO: 130, CDR-L2 set forth in SEQ ID NO: 131, and CDR-L3 set forth in SEQ ID NO: 132. In any of the provided embodiments, one of the first or second antigen-binding domains comprises a VH region and a VL region comprising the amino acid sequences set forth in SEQ ID NOs: 27 and 28, respectively. In any of the provided embodiments, one of the first antigen-binding domain or the second antigen-binding domain comprises the amino acid sequence set forth in SEQ ID NO:8, or an amino acid sequence exhibiting at least or at least about 90%, at least or at least about 90%, at least or at least about 91%, at least 92% or at least about 92%, at least 93% or at least about 93%, at least 94% or at least about 94%, at least 95% or at least about 95%, at least 96% or at least about 96%, at least 97% or at least about 97%, at least 98% or at least about 98%, or at least 99% or at least about 99% sequence identity to SEQ ID NO:8.

[0159] In any of the provided embodiments, one of the first antigen-binding domain or the second antigen-binding domain comprises a VH region comprising CDR-H1 set forth in SEQ ID NO:209, CDR-H2 set forth in SEQ ID NO:210, and CDR-H3 set forth in SEQ ID NO:211, and a VL region comprising CDR-L1 set forth in SEQ ID NO:230, CDR-L2 set forth in SEQ ID NO:231, and CDR-L3 set forth in SEQ ID NO:232; and the other of the first antigen-binding domain or the second antigen-binding domain comprises CDR-H1 set forth in SEQ ID NO:125, CDR-H2 set forth in SEQ ID NO:126, and CDR-H3 set forth in SEQ ID NO:127, and CDR-L1 set forth in SEQ ID NO:130, CDR-L2 set forth in SEQ ID NO:131, and CDR-L3 set forth in SEQ ID NO:132. In some of the provided embodiments, one of the first antigen-binding domain or the second antigen-binding domain comprises a VH region and a VL region comprising the amino acid sequences set forth in SEQ ID NO: 197 and 198, respectively; and the other of the first antigen-binding domain or the second antigen-binding domain comprises a VH region and a VL region comprising the amino acid sequences set forth in SEQ ID NO: 27 and 28, respectively. In some of the provided embodiments, one of the first antigen-binding domain or the second antigen-binding domain comprises the amino acid sequence set forth in SEQ ID NO: 241, and the other of the first antigen-binding domain or the second antigen-binding domain comprises the amino acid sequence set forth in SEQ ID NO: 8.

[0160] In some of any of the provided embodiments, one of the first antigen-binding domain or the second antigen-binding domain is encoded by the nucleotide sequence set forth in SEQ ID NO: 310. In some of any of the provided embodiments, one of the first antigen-binding domain or the second antigen-binding domain is encoded by the nucleotide sequence set forth in SEQ ID NO: 264 or SEQ ID NO: 311. In some of any of the provided embodiments, the first antigen-binding domain or the second antigen-binding domain is encoded by the nucleotide sequence set forth in SEQ ID NO: 310, and the other of the first antigen-binding domain or the second antigen-binding domain is encoded by the nucleotide sequence set forth in SEQ ID NO: 311.

[0161] In some of any of the provided embodiments, (b) is or comprises a spacer. In some of any of the provided embodiments, (b) comprises a portion of an immunoglobulin. In some of any of the provided embodiments, (b) comprises sequences of a hinge region, a CH2 region, and a CH3 region. In some of any of the provided embodiments, the hinge region comprises all or a portion of an IgG4 hinge region and / or an IgG2 hinge region, wherein the IgG4 hinge region is optionally a human IgG4 hinge region and the IgG2 hinge region is optionally a human IgG2 hinge region; C H 2 regions, IgG4 C H 2 and / or IgG2 C H 2, and the IgG4 C H 2 is optionally human IgG4 C H 2, and the IgG2 C H 2 is optionally human IgG2 C H 2; and / or C H The three regions are IgG4 C H 3 and / or IgG2 C H 3, and the IgG4 C H 3 is optionally human IgG4 C H3, and the IgG2 C H 3 is optionally human IgG2 C H In some of any of the provided embodiments, the hinge region, CH2, and CH3, are all or a portion of a hinge from human IgG4, C H All or part of 2, and C H In some of the embodiments provided, the hinge region, C H 2, and C H 3 is chimeric, and includes a hinge derived from human IgG4 and human IgG2, C H 2, and C H In some of any of the provided embodiments, (b) is selected from the group consisting of an IgG4 / 2 chimeric hinge region, or a modified IgG4 hinge region comprising at least one amino acid substitution compared to a human IgG4 hinge; an IgG2 / 4 chimeric hinge region; H 2 region; and IgG4 C H Includes three areas.

[0162] In some of any of the provided embodiments, (b) is or comprises a spacer. In some of any of the provided embodiments, (b) is 125 or about 125 to 300 or about 300 amino acids in length, 125 or about 125 to 250 or about 250 amino acids in length, 125 or about 125 to 230 or about 230 amino acids in length, 125 or about 125 to 200 or about 200 amino acids in length, 125 or about 125 to 180 or about 180 amino acids in length, 125 or about 125 to 150 or is about 150 amino acids in length, 150 or about 150 to 300 or about 300 amino acids in length, 150 or about 150 to 250 or about 250 amino acids in length, 150 or about 150 to 230 or about 230 amino acids in length, 150 or about 150 to 200 or about 200 amino acids in length, 150 or about 150 to 180 or about 180 amino acids in length, 180 or about 180 to 300 or about 300 amino acids in length, 180 or Approximately 180 to 250 or approximately 250 amino acids in length, 180 or approximately 180 to 230 or approximately 230 amino acids in length, 180 or approximately 180 to 200 or approximately 200 amino acids in length, 200 or approximately 200 to 300 or approximately 300 amino acids in length, 200 or approximately 200 to 250 or approximately 250 amino acids in length, 200 or approximately 200 to 230 or approximately 230 amino acids in length, 230 or approximately 230 to 300 or approximately 300 amino acids in length In some embodiments, (b) is 230 or about 230 to 250 or about 250 amino acids in length, or 250 or about 250 to 300 or about 300 amino acids in length, and optionally the spacer is 224 or about 224 amino acids in length, 225 or about 225 amino acids in length, 226 or about 226 amino acids in length, 227 or about 227 amino acids in length, 228 or about 228 amino acids in length, or 229 or about 229 amino acids in length. In some of any of the provided embodiments, (b) is or comprises the amino acid sequence set forth in SEQ ID NO:17.In some of any of the provided embodiments, one of the first CAR or the second CAR (b) is encoded by the nucleotide sequence set forth in SEQ ID NO:48, and the other of the first CAR or the second CAR (b) is encoded by the nucleotide sequence set forth in SEQ ID NO:305.

[0163] In some of any of the provided embodiments, (c) is or comprises a transmembrane domain. In some of any of the provided embodiments, (c) is or comprises a transmembrane domain of CD4, CD28, or CD8, optionally a transmembrane domain derived from human CD4, human CD28, or human CD8. In some of any of the provided embodiments, (c) is or comprises a transmembrane domain of human CD28. In some of any of the provided embodiments, (c) is or comprises the amino acid sequence set forth in SEQ ID NO:18. In some of any of the provided embodiments, (c) of one of the first CAR or the second CAR is encoded by the nucleotide sequence set forth in SEQ ID NO:56, and (c) of the other of the first CAR or the second CAR is encoded by the nucleotide sequence set forth in SEQ ID NO:307.

[0164] In some of any of the provided embodiments, (d) is or comprises an intracellular signaling region comprising an intracellular signaling domain and a costimulatory signaling region. In some of any of the provided embodiments, the intracellular signaling domain of (d) is capable of inducing a primary activation signal in a T cell, is a component of a T cell receptor (TCR), and / or comprises an immunoreceptor tyrosine-based activation motif (ITAM). In some of any of the provided embodiments, the intracellular signaling domain of (d) is or comprises the cytoplasmic signaling domain of the CD3-zeta (CD3ζ) chain or a functional variant or signaling portion thereof, optionally the cytoplasmic signaling domain of the human CD3 zeta chain. In some of any of the provided embodiments, the intracellular signaling domain of (d) is or comprises the amino acid sequence set forth in SEQ ID NO:20. In some of any of the provided embodiments, the intracellular signaling domain of (d) of one of the first CAR or the second CAR is encoded by the nucleotide sequence set forth in SEQ ID NO:58, and the intracellular signaling domain of (d) of the other of the first CAR or the second CAR is encoded by the nucleotide sequence set forth in SEQ ID NO:309. In some of any of the provided embodiments, (d) is or comprises an intracellular signaling region comprising an intracellular signaling domain and a costimulatory signaling region. In some of any of the provided embodiments, the costimulatory signaling region of (d) comprises the intracellular signaling domain of a T cell costimulatory molecule, or a signaling portion thereof. In some of any of the provided embodiments, the costimulatory signaling region of (d) comprises the intracellular signaling domain of CD28, 4-1BB, or ICOS, or a signaling portion thereof, optionally the intracellular signaling domain of human CD28, human 4-1BB, or human ICOS. In some of any of the provided embodiments, the costimulatory signaling region of (d) comprises the intracellular signaling domain of 4-1BB.In some of any of the provided embodiments, the (d) costimulatory signaling region is or comprises the amino acid sequence set forth in SEQ ID NO: 19. In some of any of the provided embodiments, the (d) costimulatory signaling region of one of the first CAR or the second CAR is encoded by the nucleotide sequence set forth in SEQ ID NO: 60, and the (d) costimulatory signaling region of the other of the first CAR or the second CAR is encoded by the nucleotide sequence set forth in SEQ ID NO: 308.

[0165] In some of any of the provided embodiments, (a) is or comprises a first antigen-binding domain or a second antigen-binding domain, (b) is or comprises a spacer, (c) is or comprises a transmembrane domain, and (d) is or comprises an intracellular signaling region comprising an intracellular signaling domain and a costimulatory signaling region. In some of any of the provided embodiments, one of the first CAR or the second CAR comprises (a) a first antigen binding domain that binds to GPRC5D, optionally encoded by the nucleotide sequence set forth in SEQ ID NO:311, (b) a spacer encoded by nucleotides set forth in SEQ ID NO:305, (c) a transmembrane domain encoded by the nucleotide sequence set forth in SEQ ID NO:307, and (d) an intracellular signaling region comprising an intracellular signaling domain encoded by the nucleotide sequence set forth in SEQ ID NO:309 and a costimulatory signaling region encoded by the nucleotide sequence set forth in SEQ ID NO:308; and the other of the first CAR or the second CAR comprises (a) an antigen binding domain that binds BCMA, optionally encoded by the nucleotide sequence set forth in SEQ ID NO:310, (b) a spacer encoded by nucleotides set forth in SEQ ID NO:48, (c) a transmembrane domain encoded by the nucleotide sequence set forth in SEQ ID NO:307, and (d) an intracellular signaling region comprising an intracellular signaling domain encoded by the nucleotide sequence set forth in SEQ ID NO:309 and a costimulatory signaling region encoded by the nucleotide sequence set forth in SEQ ID NO:308. and (d) an intracellular signaling region comprising an intracellular signaling domain encoded by the nucleotide sequence set forth in SEQ ID NO:58 and a costimulatory signaling domain region encoded by the nucleotide sequence set forth in SEQ ID NO:60.

[0166] In some of any of the provided embodiments, a first nucleic acid sequence encoding a first CAR is located toward the 5' end of the polynucleotide relative to a second nucleic acid sequence encoding the first CAR. In some of any of the provided embodiments, the first CAR comprises an antigen binding domain that binds to GPRC5D, and the second CAR comprises an antigen binding domain that binds to BCMA. In some of any of the provided embodiments, the first CAR comprises an antigen binding domain that binds to BCMA, and the second CAR comprises an antigen binding domain that binds to GPRC5D.

[0167] Also disclosed is a polynucleotide comprising: (i) a first nucleic acid sequence encoding a first chimeric antigen receptor (CAR), (ii) a second nucleic acid sequence encoding a second chimeric antigen receptor (CAR), and (iii) a nucleotide sequence encoding a multicistronic element, wherein the first nucleic acid encoding the first CAR and the second nucleic acid encoding the second CAR are separated by the multicistronic element; the first CAR comprises a first antigen binding domain that binds to GPRC5D, the first antigen binding domain optionally encoded by the nucleotide sequence set forth in SEQ ID NO:311; a spacer encoded by nucleotides set forth in SEQ ID NO:305; a transmembrane domain encoded by the nucleotide sequence set forth in SEQ ID NO:307; and an intracellular signaling region comprising an intracellular signaling domain encoded by the nucleotide sequence set forth in SEQ ID NO:309 and a costimulatory signaling region encoded by the nucleotide sequence set forth in SEQ ID NO:308; and the second CAR comprises a second antigen binding domain that binds BCMA, optionally encoded by the nucleotide sequence set forth in SEQ ID NO:311. Also provided herein is a polynucleotide comprising: a second antigen-binding domain encoded by the nucleotide sequence set forth in SEQ ID NO:310; a spacer encoded by the nucleotides set forth in SEQ ID NO:48; a transmembrane domain encoded by the nucleotide sequence set forth in SEQ ID NO:56; and an intracellular signaling region comprising an intracellular signaling domain encoded by the nucleotide sequence set forth in SEQ ID NO:58 and a costimulatory signaling domain region encoded by the nucleotide sequence set forth in SEQ ID NO:60, wherein a first nucleic acid sequence encoding the first CAR is positioned toward the 5' end of the polynucleotide relative to a second nucleic acid sequence encoding the second CAR.

[0168] Also disclosed is a polynucleotide comprising: (i) a first nucleic acid sequence encoding a first chimeric antigen receptor (CAR), (ii) a second nucleic acid sequence encoding a second chimeric antigen receptor (CAR), and (iii) a nucleotide sequence encoding a multicistronic element, wherein the first nucleic acid encoding the first CAR and the second nucleic acid encoding the second CAR are separated by the multicistronic element; the first CAR comprises a first antigen binding domain that binds to BCMA, the first antigen binding domain optionally encoded by the nucleotide sequence set forth in SEQ ID NO:310; a spacer encoded by nucleotides set forth in SEQ ID NO:48; a transmembrane domain encoded by the nucleotide sequence set forth in SEQ ID NO:56; and an intracellular signaling region comprising an intracellular signaling domain encoded by the nucleotide sequence set forth in SEQ ID NO:58 and a costimulatory signaling domain region encoded by the nucleotide sequence set forth in SEQ ID NO:60; and the second CAR comprises a second antigen binding domain that binds to GPRC5D, optionally encoded by the nucleotide sequence set forth in SEQ ID NO: Also provided herein is a polynucleotide comprising: a second antigen-binding domain encoded by the nucleotide sequence set forth in SEQ ID NO:311; a spacer encoded by the nucleotides set forth in SEQ ID NO:305; a transmembrane domain encoded by the nucleotide sequence set forth in SEQ ID NO:307; and an intracellular signaling region comprising an intracellular signaling domain encoded by the nucleotide sequence set forth in SEQ ID NO:309 and a costimulatory signaling region encoded by the nucleotide sequence set forth in SEQ ID NO:308, wherein a first nucleic acid encoding the first CAR is positioned toward the 5' end of the polynucleotide relative to a second nucleic acid sequence encoding the second CAR.

[0169] In some of any of the provided embodiments, the multicistronic element comprises the amino acid sequence set forth in SEQ ID NO:37. In some of any of the provided embodiments, the multicistronic element is encoded by the nucleotide sequence set forth in SEQ ID NO:44 or SEQ ID NO:45. In some of any of the provided embodiments, the multicistronic element is encoded by the nucleotide sequence set forth in SEQ ID NO:44. In some of any of the provided embodiments, the multicistronic element is encoded by the nucleotide sequence set forth in SEQ ID NO:45. In some of any of the provided embodiments, the multicistronic element is encoded by the nucleotide sequence set forth in SEQ ID NO:319.

[0170] In some of any of the embodiments provided, the polynucleotide comprises the nucleotide sequence set forth in SEQ ID NO: 299. In some of any of the embodiments provided, the polynucleotide encodes the sequence set forth in SEQ ID NO: 298.

[0171] In some of any of the embodiments provided, the polynucleotide comprises the nucleotide sequence set forth in SEQ ID NO: 302. In some of any of the embodiments provided, the polynucleotide encodes the sequence set forth in SEQ ID NO: 301.

[0172] In some of any of the embodiments provided, the polynucleotide comprises the nucleotide sequence set forth in SEQ ID NO: 315. In some of any of the embodiments provided, the polynucleotide comprises the nucleotide sequence set forth in SEQ ID NO: 316.

[0173] Also provided herein is a polynucleotide encoding a GPRC5D-binding domain, a BCMA-binding domain, and an intracellular signaling region comprising the intracellular signaling domain of 4-1BB. In some of any of the provided embodiments, the polynucleotide comprises the nucleotide sequence set forth in SEQ ID NO:317.

[0174] Also provided are vectors comprising any of the provided polynucleotides. In some of any of the provided embodiments, the vector is a viral vector. In some of any of the provided embodiments, the viral vector is a lentiviral vector or a retroviral vector.

[0175] Also provided are modified cells comprising any of the chimeric antigen receptors provided herein. In some of any of the provided embodiments, the modified cells comprise a chimeric antigen receptor provided herein and further comprise a polynucleotide comprising a nucleotide sequence encoding a second chimeric antigen receptor.

[0176] Also provided are modified cells comprising any of the polynucleotides provided herein.

[0177] In some of any of the provided embodiments, the modified cells are lymphocytes. In some of any of the provided embodiments, the modified cells are NK cells or T cells. In some of any of the provided embodiments, the modified cells are T cells, and the T cells are CD4+ T cells or CD8+ T cells.

[0178] In some of any of the provided embodiments, the modified cells are modified primary cells obtained from the subject.

[0179] In some of any of the provided embodiments, the modified cells are such that of a plurality of said modified cells, less than or about 10%, less than or about 9%, less than or about 8%, less than or about 7%, less than or about 7%, less than or about 5%, less than or about 4%, less than or about 4%, less than or about 3%, less than or about 2%, or less than or about 1% of the cells of said plurality comprise a chimeric antigen receptor that exhibits tonic signaling and / or antigen-independent activity or signaling.

[0180] Also provided are compositions containing any of the chimeric receptors provided herein. In some of any of the provided embodiments, the compositions comprise CD4+ T cells and CD8+ T cells, and the ratio of CD4+ T cells to CD8+ T cells is 1:3 to 3:1 or about 1:3 to about 3:1. In some embodiments, the ratio of CD4+ T cells to CD8+ T cells in the composition is 1:2 to 2:1. In some embodiments, the ratio of CD4+ T cells to CD8+ T cells in the composition is 1:1. In some of any of the provided embodiments, the composition further comprises a pharmaceutically acceptable excipient. In some of any of the provided embodiments, the composition is sterile.

[0181] Also provided herein are methods of treatment that include administering to a subject having a disease or disorder any of the compositions provided herein comprising any of the modified cells provided herein or any of the compositions provided herein comprising any of the chimeric antigen receptors provided herein. In some of any of the provided embodiments, the dose of cells is 2.5 x 10 7 Or about 2.5 x 10 7 CAR-expressing T cells ~1.2 x 10 9 or about 1.2 x 10 9 CAR-expressing T cells, 5.0 x 10 7 or approximately 5.0 x 10 7 CAR-expressing T cells ~4.5 x 10 8 or about 4.5 x 108 CAR-expressing T cells, 1.5 x 10 8 Or about 1.5 x 10 8 CAR-expressing T cells ~3.0 x 10 8 Or about 3.0 x 10 8 In some of either embodiment, the dose of cells comprises 1 x 10 CAR-expressing T cells. 7 Or about 1 x 10 7 CAR-expressing T cells ~2 x 10 9 Or about 2 x 10 9 In some of either embodiment, the dose of cells comprises 2.5 x 10 CAR-expressing T cells. 7 Or about 2.5 x 10 7 CAR-expressing T cells ~1.2 x 10 9 or about 1.2 x 10 9 CAR-expressing T cells, 5.0 x 10 7 or approximately 5.0 x 10 7 CAR-expressing T cells ~4.5 x 10 8 or about 4.5 x 10 8 CAR-expressing T cells, or 1.5 x 10 8 Or about 1.5 x 10 8 CAR-expressing T cells ~3.0 x 10 8 Or about 3.0 x 10 8 In some of either embodiment, the dose of cells comprises 1.5 x 10 CAR-expressing T cells. 7 Or about 1.5 x 10 7 pieces, 2.5×10 7 Or about 2.5 x 10 7 pieces, 5.0×10 7 or approximately 5.0 x 10 7 pieces, 7.5×10 7 Or about 7.5 x 10 7 pieces, 1.5×10 8 Or about 1.5 x 10 8 pieces, 2.25×10 8 Or about 2.25 x 10 8 pieces, 3.0×10 8 Or about 3.0 x 10 8 pieces, 4.5×10 8 or about 4.5 x 10 8pieces, 6.0×10 8 Or about 6.0 x 10 8 pieces, 8.0×10 8 or approximately 8.0 x 10 8 pieces, or 1.2 x 10 9 or about 1.2 x 10 9 In some of either embodiment, the dose of cells comprises 5.0 x 10 CAR-expressing T cells. 7 or approximately 5.0 x 10 7 pieces, 1.5×10 8 Or about 1.5 x 10 8 pieces, 3.0×10 8 Or about 3.0 x 10 8 or 4.5 x 10 8 or about 4.5 x 10 8 In some of either embodiment, the dose of cells comprises 5.0 x 10 CAR-expressing T cells. 7 or approximately 5.0 x 10 7 pieces, 1.5×10 8 Or about 1.5 x 10 8 pieces, 3.0×10 8 Or about 3.0 x 10 8 pieces, or 4.5 x 10 8 or about 4.5 x 10 8 In some of any of the embodiments, the dose of cells comprises 5.0 x 10 CAR-expressing T cells. 7 or approximately 5.0 x 10 7 Contains CAR-expressing T cells.

[0182] Also provided herein are methods of treatment that include administering to a subject having a disease or disorder any of the compositions provided herein comprising any of the modified cells provided herein or any of the compositions provided herein comprising any of the chimeric antigen receptors provided herein. In some of any of the provided embodiments, the dose of cells is 1.0 x 10 7 or approximately 1.0 x 10 7 CAR-expressing T cells ~1.2 x 10 9 or about 1.2 x 10 9 CAR-expressing T cells, 1.5 x 107 Or about 1.5 x 10 7 CAR-expressing T cells ~4.5 x 10 8 or about 4.5 x 10 8 CAR-expressing T cells, 2.0 x 10 7 Or about 2.0 x 10 7 CAR-expressing T cells ~3.0 x 10 8 Or about 3.0 x 10 8 Contains CAR-expressing T cells.

[0183] Also provided herein are methods of treatment comprising administering to a subject having a disease or disorder a composition containing a plurality of modified cells comprising a first chimeric antigen receptor and a second chimeric antigen receptor, each of which is any chimeric antigen receptor provided herein or any chimeric antigen receptor encoded by any polynucleotide provided herein; and administering to the subject a composition containing a plurality of second modified cells comprising the second chimeric antigen receptor. In some of any of the provided embodiments, the dose of the first plurality of modified cells and the dose of the second plurality of modified cells are independently 1.0 x 10 7 or approximately 1.0 x 10 7 CAR-expressing T cells ~1.5 x 10 9 Or about 1.5 x 10 9 CAR-expressing T cells, 1.25 x 10 7 Or about 1.25 x 10 7 CAR-expressing T cells ~0.6 x 10 8 or approximately 0.6 x 10 8 CAR-expressing T cells, 2.5 x 10 7 Or about 2.5 x 10 7 CAR-expressing T cells ~2.25 x 10 8 Or about 2.25 x 10 8 CAR-expressing T cells, 7.5 x 10 7 Or about 7.5 x 10 7 CAR-expressing T cells ~1.5 x 10 8 Or about 1.5 x 10 8 CAR-expressing T cells, 2.5 x 10 7Or about 2.5 x 10 7 CAR-expressing T cells ~1.2 x 10 9 or about 1.2 x 10 9 CAR-expressing T cells, 5.0 x 10 7 or approximately 5.0 x 10 7 CAR-expressing T cells ~4.5 x 10 8 or about 4.5 x 10 8 CAR-expressing T cells, 1.5 x 10 8 Or about 1.5 x 10 8 CAR-expressing T cells ~3.0 x 10 8 Or about 3.0 x 10 8 Contains CAR-expressing T cells.

[0184] In some of any of the provided embodiments of the provided methods, the disease or disorder is associated with expression of G protein-coupled receptor class C group 5 member D (GPRC5D).

[0185] In some of any of the provided embodiments of the provided methods, the disease or disorder is further associated with expression of B-cell maturation antigen (BCMA).

[0186] In some of any of the provided embodiments of the provided methods, the disease or disorder is a B-cell associated disorder. In some of any of the provided embodiments of the provided methods, the disease or disorder associated with BCMA is an autoimmune disease or disorder. In some of any of the provided embodiments of the provided methods, the autoimmune disease or disorder is systemic lupus erythematosus (SLE), lupus nephritis, inflammatory bowel disease, rheumatoid arthritis, ANCA-associated vasculitis, idiopathic thrombocytopenic purpura (ITP), thrombotic thrombocytopenic purpura (TTP), autoimmune thrombocytopenia, Chagas' disease, Graves' disease, Wegener's granulomatosis, polyarteritis nodosa, Sjogren's syndrome, pemphigus vulgaris, scleroderma, multiple sclerosis, psoriasis, IgA nephropathy, IgM polyneuropathy, vasculitis, diabetes mellitus, Raynaud's syndrome, antiphospholipid syndrome, Goodpasture's disease, Kawasaki's disease, autoimmune hemolytic anemia, myasthenia gravis, or progressive glomerulonephritis.

[0187] In some of any of the provided embodiments of the provided methods, the disease or disorder is cancer. In some of any of the provided embodiments of the provided methods, the cancer is a GPRC5D-expressing cancer. In some of any of the provided embodiments of the provided methods, the cancer is a plasma cell malignancy, and the plasma cell malignancy is multiple myeloma (MM) or plasmacytoma. In some of any of the provided embodiments of the provided methods, the cancer is multiple myeloma (MM). In some of any of the provided embodiments of the provided methods, the cancer is relapsed / refractory multiple myeloma.

[0188] In some of any of the provided embodiments of the provided methods, the subject is refractory to BCMA-targeted therapy, optionally to T cells comprising a CAR that specifically binds BCMA, or has relapsed after administration of BCMA-targeted therapy, optionally after administration of T cells comprising a CAR that specifically binds BCMA. In some of any of the provided embodiments of the provided methods, a subject who is refractory to BCMA-targeted therapy, optionally to T cells comprising a CAR that specifically binds BCMA, or has relapsed after administration of BCMA-targeted therapy, optionally after administration of T cells comprising a CAR that specifically binds BCMA, is selected for treatment. In some of any of the provided embodiments of the provided methods, the subject has previously undergone administration of a BCMA-targeted therapy for the treatment of the disease or disorder prior to administration of the dose of cells. In some of any of the provided embodiments of the provided methods, the subject has previously undergone administration of a BCMA-targeted therapy for the treatment of the disease or disorder prior to administration of the first dose of cells and the second dose of cells.

[0189] In some of any of the provided embodiments of the provided methods, the BCMA-targeted therapy comprises a composition comprising T cells comprising a CAR that specifically binds BCMA. In some of any of the provided embodiments of the provided methods, the subject is refractory to the BCMA-targeted therapy, optionally to T cells comprising a CAR that specifically binds BCMA, or has relapsed after administration of the BCMA-targeted therapy, optionally after administration of T cells comprising a CAR that specifically binds BCMA. In some of any of the provided embodiments of the provided methods, the subject comprises multiple myeloma cells that exhibit loss of the BCMA antigen or epitope, downregulation of BCMA, and / or BCMA-negative tumor cells after the previous administration. [Brief explanation of the drawings]

[0190] [Figure 1A]Figure 1A shows CD138 mRNA expression data (log2 scale) from the Cancer Cell Line Encyclopedia. Cancer types, from left to right: upper aerodigestive (32); esophageal (25); prostate (7); multiple myeloma (30); bile duct (8); lung (131); pancreas (44); kidney (34); breast (58); colorectal (61); stomach (38); meningioma (3); liver (28); glioma (62); osteosarcoma (10); thyroid (12); uterine mucosa (27); soft tissue (21); mesothelioma (11); ovarian (51); chondrosarcoma (4); small cell lung (53); melanoma (61); neuroblastoma (17); medulloblastoma (4); Ewing's sarcoma (12); Hodgkin's lymphoma (12); DLBCL (18); other lymphomas (28); B-cell (15); CML (15); Burkitt's lymphoma (11); T-cell (16); AML (34); other leukemia (1). [Figure 1B] Figure 1B shows the mRNA expression of GPRC5D in malignant cell lines [n = 1036; Cancer Cell Line Encyclopedia (CCLE)]. RMA, robust multiarray average; DLBCL, diffuse large B-cell lymphoma; CML, chronic myeloid leukemia; ALL, acute lymphoblastic leukemia; AML, acute myeloid leukemia; NSC, non-small cell lung cancer. Cancer types, from left to right, are: multiple myeloma (30); other leukemia (1); DLBCL (18); CML (15); meningioma (3); other lymphoma (28); Burkitt lymphoma (11); Hodgkin lymphoma (12); T-cell (16); B-cell (15); biliary (8); AML (34); pancreatic (44); thyroid (12); colorectal (61); renal (34); and osteosarcoma (10). ;Urinary tract (27);Breast (58);Neuroblastoma (17);Non-small cell lung (131);Ewing's sarcoma (12);Prostate (7);Melanoma (61);Upper aerodigestive tract (32);Uterine mucosa (27);Medulloblastoma (4);Liver (28);Ovary (51);Stomach (38);Glioma (62);Small cell lung (53);Mesothelioma (11);Esophagus (25);Other (150);Chondrosarcoma (4). [Figure 2A]Figure 2A shows GTEx RNASeq expression data for CD138 in various organs. Tissue types, from left to right, are: cerebellum, cerebral hemispheres, anterior cingulate cortex, frontal cortex, cortex, amygdala, hippocampus, nucleus accumbens, caudate nucleus, putamen, sigmoid colon, tibial nerve, skeletal muscle, uterus, esophageal muscularis, esophagogastric junction of esophagus, hypothalamus, adipose tissue, cervical tissue, coronary artery, cervical spinal cord, substantia nigra, ovary, tibial artery, mammary tissue, fallopian tube, adipose tissue, kidney, left ventricle, neck, adrenal gland, bladder, whole blood, skin (sun-exposed), skin (unexposed), aorta, tonsil, small intestine, pancreas, liver, atrial appendage, vagina, stomach, prostate, spleen, umbilical cord blood, thyroid gland, transverse colon, pituitary gland, esophageal mucosa, testis, minor salivary gland, lung, and bone marrow. [Figure 2B] Figure 2B shows the mRNA expression levels of GPRC5D in normal tissues based on Gtex RNASeq data (Gtex 708 ENSG00000111291.4). The dotted line represents the GPRC5D expression level in CD138-sorted primary MM cells (Blueprint RNAseq; n=9). FPKM, fragments per kilobase of transcript per million mapped reads. Tissue types, from left to right: cerebellum, cerebral hemispheres, anterior cingulate cortex, frontal cortex, cortex, amygdala, hippocampus, nucleus accumbens, caudate nucleus, putamen, sigmoid colon, tibial nerve, skeletal muscle, uterus, esophageal muscularis, esophagogastric junction of esophagus, hypothalamus, adipose tissue, cervical tissue, coronary artery, cervical spinal cord, substantia nigra, ovary, tibial artery, mammary tissue, fallopian tube, adipose tissue, kidney, left ventricle, neck tissue, adrenal gland, bladder, whole blood, skin (sun exposed), skin (unexposed), aorta, tonsil, small intestine, pancreas, liver, auricle, vagina, stomach, prostate, spleen, umbilical cord blood, thyroid gland, transverse colon, pituitary gland, esophageal mucosa, testis, minor salivary gland, lung, and primary MM (bone marrow). [Figure 2C] Figure 2C shows GPRC5D mRNA expression levels by Blueprint RNAseq in human primary tissue cell types. FPKM, fragments per kilobase transcript per million mapped reads. [Figure 3A]Figure 3A shows Kaplan-Meier curves for progression-free survival (PFS) stratified for subjects with MM whose GPRC5D expression levels by RNA-seq were either above (>) or below (<) the median GPRC5D expression level. Significance was determined by the equal hazards log-rank test (p=0.0031; n=765). [Figure 3B] Figure 3B shows the International Staging System (ISS) scores of subjects with MM stratified by GPRC5D expression level (n=369 above the median, n=374 below the median). [Figure 3C] Figures 3C-3H show the frequency of common cytogenetic abnormalities among MM subjects stratified by GPRC5D expression level (n=287-291 above the median, n=280-282 below the median). [Figure 3D] See legend to Figure 3C. [Figure 3E] See legend to Figure 3C. [Figure 3F] See legend to Figure 3C. [Figure 3G] See legend to Figure 3C. [Figure 3H] See legend to Figure 3C. [Figure 4A] Figure 4A shows boxplot quantification of GPRC5D protein outliers in cell lines after immunohistochemical detection. Boxplots of outliers show the median and interquartile range (IQR) of intramembrane optical density; whiskers are 1.5 × IQR. The mean fluorescence intensity (MFI) of GPRC5D expression in K562 cells engineered to express the protein is shown by the number following the cell line designated K562-GPRC5D. [Figure 4B] Figure 4B shows automated quantitative immunofluorescence on 83 bone marrow samples from MM patients, with each column representing an individual patient sample. [Figure 4C] Figure 4C shows the percentage of patient samples in which greater than 50% of CD138+ cells expressed BCMA, GPRC5D, or BCMA or GPRC5D as measured by automated quantitative immunofluorescence in 83 bone marrow samples from MM patients. [Figure 4D]Figure 4D shows the correlation between BCMA and GPRC5D expression in CD138+ cells; R2 = 0.156. [Figure 5] FIG. 5 shows the linear, conformational, and discontinuous epitope binding of a subset of scFvs targeting GPRC5D assessed by ELISA-based techniques. [Figure 6A] Figures 6A and 6B show antigen-independent (tonic) signaling of CARs containing the indicated scFvs and spacers. Jurkat Nur77-RFP reporter cells were transduced with one of 42 CAR / GFP bicistronic constructs. 5 x 10 viable GFP Jurkat cells were plated and monitored for RFP expression in the absence of target antigen 11 days post-transduction. Expression of both RFP and GFP indicated tonic signaling; expression of GFP alone indicated CAR transduction without tonic signaling. [Figure 6B] See legend to Figure 6A. [Figure 6C] Figures 6C-6E show antigen-dependent versus antigen-independent signaling of candidate CARs with long (Figure 6C), medium (Figure 6D), and short (Figure 6E) spacers measured after 20 hours of 2:1 culture of Jurkat Nur77-RFP reporter cells with MM.1S cells (expressing endogenous GPRC5D). Percent CAR T cell signaling is determined by RFP+GFP+ / total GFP+ cells. Representative data from two experiments. [Figure 6D] See legend to Figure 6C. [Figure 6E] See legend to Figure 6C. [Figure 6F] Figure 6F shows CAR-transduced cells designated as GFP+ on the y-axis. RFP, a surrogate for Nur77 expression, is shown on the x-axis. The percentage shown is of only transduced GFP+ cells that are RFP+ (top quadrant only). [Figure 7A]Figure 7A shows the binding of HEK293 cells transiently expressing a human G protein-coupled receptor (GPCR) library with cytoplasmic GFP to co-cultured HEK293 cells (both in suspension) transiently expressing the long spacer anti-GPRC5D scFv clone 203 and cytoplasmic mCherry 761, as quantified by automated flow cytometry analysis. Prespecified significance threshold (red line): Z-score 3; p<0.0027. [Figure 7B] Figure 7B shows binding of anti-GPRC5D scFv clone 203 mIgG2a Fc chimeric antibody to HEK293 cells expressing the indicated cell surface proteins. Confirmation of binding to potential off-target proteins and nonspecific binders identified in a microarray screen of >4,400 transmembrane proteins is shown. ZsGreen1: transfection control; isotype: irrelevant scFv-mIgG2a Fc negative control; CTLA-4 / CD86 interaction: positive control. [Figure 7C] Figure 7C shows the results of assessing the activation of potential off-target proteins PCDH1A or FCGR2A via GPRC5D(203)CAR. Jurkat Nur77-RFP-activated reporter cells expressing a bicistronic plasmid containing GPRC5D(203)CAR and GFP were co-cultured with K562 cells expressing the indicated antigens, GPRC5D (positive control) or BCMA (negative control). Activation was determined as % RFP+GFP+ / total GFP+ cells. [Figure 7D] Figure 7D shows that CRISPR-Cas9-mediated knockout of GPRC5D from MM cell lines abolished activation of GPRC5D(203)CAR-Jurkat Nur77 reporter cells, as assessed by flow cytometric measurement of changes in RFP expression levels. [Figure 8A] FIG. 8A shows the expression levels of GPRC5D mRNA in MM cell lines and primary MM cells (boxed). [Figure 8B]Figure 8B shows the results of cytotoxicity of GPRC5D(203)-expressing CAR T cells against MM1.S, OPM2, and RPMI-8226 target cells after 24 hours of co-culture, normalized to donor-matched mock-transduced CAR T cells and shown by percent lysis (technical triplicates for each of the two donors; mean ± SD). [Figure 9A] Figure 9A shows cell killing of OPM2-ffLuc MM cells induced by CAR T cells incorporating the indicated scFvs after 24 hours of co-culture, as indicated by ATP-dependent bioluminescence after addition of luciferin; normalized to tumor cell-only controls (pooled data of two experiments each performed in triplicate, mean ± SEM; p<0.001). [Figure 9B] Figures 9B and 9C show flow cytometry analysis demonstrating killing of primary bone marrow mononuclear cells (BMMCs) from a patient with multiple relapsed MM after overnight co-culture with anti-GPRC5D CAR T cells at a 1:1 ratio of CAR+ T cells:BMMCs. MM cells, CD138+ / CD38hi; plots gated on viable CD3- cells. [Figure 9C] See legend to Figure 9B. [Figure 9D] FIG. 9D shows flow cytometry analysis of primary BMMCs from additional patients plotted for CD138+ / CD3−. [Figure 10A] Figures 10A-10C show cytokines produced by CAR T cells incorporating the indicated scFvs, measured in supernatants by multiplex Luminex assay, after 24 hours of culture alone or 1:1 with OPM2 MM cells. [Figure 10B] See legend to Figure 10A. [Figure 10C] See legend to Figure 10A. [Figure 11A]Figures 11A and 11B show the proliferation of mock-transduced or GPRC5D(203)-expressing CAR T cells cultured alone or with B-ALL (Nalm6; GPRC5D-) or MM (OPM2; endogenous GPRC5D+) cells at a 1:1 ratio, and Figures 11C and 11D show their activation. T cells were stained with CellTrace Violet (CTV) before co-culture and stained for CD4, CD8, and CD25 72 hours later. (A,B) Proliferation is indicated by a decrease in CTV fluorescence. (C,D) Activation is indicated by an increase in CD25 fluorescence. [Figure 11B] See legend to Figure 11A. [Figure 11C] See legend to Figure 11A. [Figure 11D] See legend to Figure 11A. [Figure 12A] Figure 12A shows a representative FACS analysis of CAR expression levels in CAR T cells measured using an antibody specific for the spacer. [Figure 12B] Figure 12B shows the survival rate of mice treated with 3x106 4-1BB-containing CAR T cells incorporating the indicated anti-GPRC5D scFv clones 14 days after OPM2 injection (n=8 / arm). [Figure 12C] Figure 12C shows tumor volume and survival rates of mice in the RPMI-8226 xenograft model from one of two experiments; median survival 29 days vs. 50 days (p<0.05; n=5 / arm, selected figures from two experiments). [Figure 12D] Figure 12D shows tumor volume and CAR T cell expansion in mice in the RPMI-8226 xenograft model, monitored by flow cytometry of peripheral blood using an antibody against the spacer to detect the CAR (p<0.001; n=10 / arm; both time points). [Figure 13A]Figure 13A shows the survival of mice treated 21 days after OPM2 injection with 3x10 T cells genetically engineered to express CARs incorporating bicistronic constructs encoding extGLuc and scFv CD19 (SJ25C1) or GPRC5D (203) and either 4-1BB or CD28 costimulatory domains (n=5 / arm). [Figure 13B] Figures 13B, C, and D show tumor burden (D-luciferin bioluminescence imaging [BLI] of OPM-ffLuc) in the mice in Figure 13A. [Figure 13C] See legend to Figure 13B. [Figure 13D] See legend to Figure 13B. [Figure 13E] Figure 13E shows the results of CAR T cell homing (coelenterazine BLI of extGLuc CAR T cells) of the mice in Figure 13A performed 7 days after CAR T cell treatment. [Figure 14A] Figures 14A and 14B show the dose response of GPRC5D-targeted CAR T cell therapy and BCMA-targeted CAR T cell therapy performed 14 days after OPM2 injection (n=8 mice / arm). Figure 14A shows the tumor burden assessed by BLI of OPM-ffLuc. Figure 14B shows the survival fraction (p values ​​shown are relative to mock-transduced or irrelevant targeting CAR T cells). [Figure 14B] See legend to Figure 14A. [Figure 15A] Figures 15A-15C show the levels of IFN-gamma (Figure 15A), TNF-alpha (Figure 15B), and IL-2 (Figure 15C) after 20 hours of co-culture of GPRC5D(203), anti-BCMA, or mock-treated T cells with 20 different normal human primary cell types or OPM2 cells (mean ± SD). [Figure 15B] See legend to Figure 15A. [Figure 15C] See legend to Figure 15A. [Figure 15D]Figure 15D shows the results of screening mouse and cynomolgus monkey cross-reactive scFv clones for tonic signaling. % RFP+ indicates activation after co-culture at a 1:1 effector:target ratio (compared to GFP+ CAR-transduced cells). [Figure 16A-B] Figures 16A-C show the body mass change (Figure 16A), body temperature (Figure 16B) or BLI of OPM2-ffLuc cells (Figure 16C) after injection of mice with 3 x 10 human T cells expressing a CAR containing a human / mouse cross-reactive anti-GPRC5D scFv (clone 205). [Figure 16C] See legend to Figures 16A-B. [Figure 17A] Figure 17A shows a representative FACs analysis of CAR expression levels measured using truncated receptor surrogate markers in non-human primate (NHP) T cells transduced to express either cynomolgus cross-reactive GPRC5D CAR or cynomolgus GPRC5D. [Figure 17B] Figure 17B shows target lysis, and Figure 17C shows IFNγ production by mock T cells against NHP T cells transduced to express either the cynomolgus monkey cross-reactive GPRC5D CAR or autologous target antigen-presenting cells (tAPCs) at various effector-to-target (E:T) ratios. [Figure 17C] See legend to Figure 17B. [Figure 17D] Figure 17D shows target lysis, and Figure 17E shows IFNγ production by NHP T cells transduced to express either the cynomolgus monkey cross-reactive GPRC5D CAR or mock T cells against target K562, i.e., K562-GPRC5D cells, at various effector-to-target (E:T) ratios. [Figure 17E] See legend to Figure 17D. [Figure 18A] Figure 18A shows the results of PCR of DNA encoding the CAR as a measure of CAR T cell persistence in peripheral blood and bone marrow 21 days post-infusion. CAR-transduced NHP T cells were used as a positive control. [Figure 18B] Figures 18B-D show the results of pathological assessments 1 to 21 days after injection of cynomolgus monkeys with cynomolgus T cells engineered to express a CAR containing the human / cynomolgus cross-reactive anti-GPRC5D scFv clone 202. Figure 18B shows body temperature, Figure 18C shows body mass change, and Figure 18D shows body mass. [Figure 18C] See legend to Figure 18B. [Figure 18D] See legend to Figure 18B. [Figure 19A] Figure 19A shows BLI images on days 7 and 15 of a mouse injected with 1 x 10 mixed population of OPM2WT and OPM2BCMA-KO (GFP / ffLuc+) cells on day 0 and 3 x 10 indicated CAR T cells on days 8 and 16, and Figure 19B shows images on day 34. n = 5 mice / arm, selected images of two experiments. [Figure 19B] See legend to Figure 19A. [Figure 20A] Figures 20A and 20B show OPM2 cells in the bone marrow of mice injected with the mixed population of OPM2 cells and CAR T cells shown in Figures 19A and 19B. Representative plots of 3 mice per arm. Live / dead gating was performed but not shown (n = 2 experiments with comparable results). [Figure 20B] See legend to Figure 20A. [Figure 21A] Figure 21A shows that tonic signaling through an exemplary anti-BCMA CAR is minimal. [Figure 21B] Figure 21B shows lysis of target cells by human primary T cells expressing this exemplary anti-BCMA CAR. [Figure 21C] Figure 21C shows the secretion of IFN-gamma by human primary T cells expressing this exemplary anti-BCMA CAR upon co-culture with target cells. [Figure 22A] FIG. 22A shows the loss of GPRC5D or BCMA expression in OPM2 cells in which GPRC5D or BCMA, respectively, was knocked out, as assessed by flow cytometry. [Figure 22B] Figure 22B shows antigen-specific activation of exemplary anti-BCMA and anti-GPRC5D CARs. [Figure 23] FIG. 23 shows the expression levels of the BCMA gene and the GPRC5D gene in multiple myeloma cell lines. [Figure 24] Figure 24 shows the expression levels of BCMA and GPRC5D proteins in multiple myeloma and control cell lines. [Figure 25A] Figures 25A and 25B show OPM2 tumor burden in mice injected with either OPM2 WT cells (Figure 25A), OPM2 BCMA KO cells (Figure 25B; upper panel), or OPM2 GPRC5D KO cells (Figure 25B; lower panel). Mice were treated with cell compositions containing cells expressing anti-BCMA CAR (BCMA) or anti-GPRC5D CAR (GPRC5D), or cell compositions containing a cell pool made to contain anti-BCMA CAR-expressing cells and anti-GPRC5D CAR-expressing cells at a 1:1 ratio (GPRC5D / BCMA pooled cells). [Figure 25B] See legend to Figure 25A. [Figure 26] Figure 26 shows the survival rate of mice injected with OPM2 tumor cells and treated with cells expressing anti-GPRC5D CAR (GPRC5D) or anti-BCMA CAR (BCMA), or a pool of anti-BCMA CAR-expressing cells and anti-GPRC5D CAR-expressing cells (GPRC5D / BCMA pooled cells) at three different doses. [Figure 27] Figure 27 shows tumor volume in mice injected with RPMI8226 cells and treated with cells expressing anti-GPRC5D CAR (GPRC5D) or anti-BCMA CAR (BCMA), or a pool of anti-BCMA CAR-expressing cells and anti-GPRC5D CAR-expressing cells (GPRC5D / BCMA pooled cells) at three different doses. [Figure 28] FIG. 28 shows the survival rate of the mice in FIG. [Figure 29]Figure 29 shows an anti-BCMA / anti-GPRC5D dual targeting strategy. (i) and (ii) represent anti-BCMA CAR-expressing cells / anti-GPRC5D CAR-expressing cell pools (GPRC5D / BCMA pool cells). (iii) and (iv) represent bicistronic constructs containing an anti-BCMA CAR and an anti-GPRC5D CAR, each separated by a self-cleaving peptide. (v) represents a "single stalk" CAR approach, in which an anti-BCMA scFv and an anti-GPRC5D scFv are in tandem, separated only by a linker. [Figure 30] FIG. 30 shows the expression of the indicated constructs on the cell surface following retroviral transduction of cells with each of the constructs in FIG. [Figure 31] FIG. 31 shows the retroviral transduction efficiency of each of the constructs shown in FIG. 29, as assessed by flow cytometry analysis. [Figure 32] Figure 32A shows the cytotoxicity of T cells expressing the constructs shown in Figure 29 when co-cultured with a wild-type OPM2 multiple myeloma cell line, as measured by the percentage of tumor cells lysed. CAR-expressing T cells and target cells were cultured at increasing E:T ratios. Figure 32B shows the cytotoxicity of cells expressing the constructs shown in Figure 29 when co-cultured with a BCMA knockout OPM2 cell line, as measured by the percentage of tumor cells lysed. CAR-expressing T cells and target cells were cultured at increasing E:T ratios. [Figure 33]Figure 33A shows the ability of T cells expressing the indicated CAR constructs to secrete various cytokines when co-cultured with target cells expressing BCMA and GPRC5D for 24 hours. Figure 33B shows the ability of T cells expressing the indicated CAR constructs to secrete various cytokines when co-cultured with BCMA-expressing GPRC5D-negative target cells for 24 hours. Figure 33C shows the ability of T cells expressing the indicated CAR constructs to secrete various cytokines when co-cultured with BCMA-expressing GPRC5D-negative target cells for 24 hours. [Figure 34] Figure 34A shows the survival of mice injected with OPM2 wild-type cells after treatment with T cells expressing the indicated CARs. Figure 34B shows the survival of the mice in Figure 34A after a second injection of BCMA knockout OPM2 cells after treatment with T cells expressing the indicated CARs. [Figure 35A] Figures 35A-C show tumor growth, assessed by bioluminescence imaging, in mice following treatment with 3x106 CAR-expressing T cells, 30 days (Figure 35A) or 105 days (Figure 35B) after the first injection of BCMA knockout OPM2 cells (2x106), or 36 days (Figure 35C) after the second injection of BCMA knockout OPM2 cells (3x106). [Figure 35B] See legend to Figure 35A. [Figure 35C] See legend to Figure 35A. [Figure 36] Figure 36 shows the survival of mice treated with a low dose (5x105) of cells expressing the indicated CARs after injection of 2x106 wild-type OPM2 cells. [Figure 37A] Figures 37A-C show tumor burden, as assessed by bioluminescence imaging, in mice injected with wild-type OPM2 cells after 0 (Figure 37A), 15 (Figure 37B), or 22 (Figure 37C) days of treatment with cells expressing the indicated CARs. [Figure 37B] See legend to Figure 37A. [Figure 37C]See legend to Figure 37A. [Figure 38] Figure 38 shows tumor burden assessed by bioluminescence imaging of wild-type OPM2 cells (left panel) and BCMA knockout OPM2 cells (right panel) in mice injected with a mixed composition of wild-type and 5-10% BCMA knockout OPM2 cells following treatment with 5x10 cells expressing the indicated CARs. [Figure 39] Figure 39 shows the survival rate of mice injected with a mixed composition of wild-type OPM2 cells and 5-10% BCMA knockout OPM2 cells following treatment with 2.5 x 10 cells expressing the indicated CARs. [Figure 40A] Figures 40A-C show tumor burden, as assessed by bioluminescence imaging, in mice injected with a mixed composition of wild-type OPM2 cells and 5-10% BCMA knockout OPM2 cells on days 0 (Figure 40A), 22 (Figure 40B), or 34 (Figure 40C) after treatment with 5 x 10 cells expressing the indicated CARs. [Figure 40B] See legend to Figure 40A. [Figure 40C] See legend to Figure 40A. [Figure 41] Figures 41A and 41B show the loss of expression of the trailing CARs (BCMA and GPRC5D, respectively) in the non-codon divergent bicistronic constructs. [Figure 42] Figures 42A and 42B show that codon divergence of the bicistronic construct rescues expression of the trailing CARs (BCMA and GPRC5D, respectively). [Figure 43] Figure 43 shows stimulation of Jurkat Nur77-RFP reporter cells expressing the indicated CARs after co-culture with target cells. [Figure 44A] Figures 44A-C show the expression levels of IFN-gamma, IL-2, and TNF-alpha (respectively) by human primary T cells expressing the indicated CARs upon co-culture with target cells. [Figure 44B]See legend to Figure 44A. [Figure 44C] See legend to Figure 44A. [Figure 45] Figure 45 shows antigen-specific activation of Jurkat Nur77-RFP reporter cells transduced with the indicated CARs when co-cultured with OPM2 WT, OPM2 BCMA KO, or OPM2 GPRC5D KO cells. [Figure 46A] Figures 46A-C show the expression of IFN-gamma, IL-2, and TNF-alpha (respectively) by human primary T cells expressing the indicated CARs when cultured with OPM2 WT, OPM2 BCMA KO, or OPM2 GPRC5D KO cells. [Figure 46B] See legend to Figure 46A. [Figure 46C] See legend to Figure 46A. [Figure 47] Figure 47A shows tumor burden (assessed by BLI) in mice injected with OPM2 WT cells and treated with cells expressing the indicated CARs. Figures 47B and 47C show tumor burden (assessed by BLI) in mice injected with a combination of OPM2 WT and BCMA KO cells (Figure 47B) or a combination of OPM2 WT and GPRC5D KO cells (Figure 47C) and treated with cells expressing the indicated CARs. [Figure 48] Figure 48 shows the survival rate (%) of the mice in Figures 47A-C. DETAILED DESCRIPTION OF THE INVENTION

[0191] Detailed Description Provided herein are G protein-coupled receptor class C group 5 member D (GPRC5D), as well as chimeric antigen receptors (CARs) that target cells expressing GPRC5D and diseases expressing GPRC5D. Also provided are cells (e.g., T cells) engineered to express the provided anti-GPRC5D CARs, and compositions containing such cells. GPRC5D is observed to be expressed, e.g., variably expressed, in certain diseases and conditions, e.g., malignant tumors, or in tissues or cells thereof, e.g., malignant plasma cells derived from relapsed myeloma patients or newly diagnosed myeloma patients, and is rarely expressed in normal tissues. Among the embodiments provided are approaches useful for treating diseases and conditions and / or targeting such cell types, e.g., nucleic acid molecules encoding receptors that bind GPRC5D, e.g., chimeric antigen receptors (CARs), and the encoded receptors, e.g., the encoded CARs, as well as compositions and articles of manufacture comprising the same. Such receptors are generally expressed by antibodies (antigen-binding antibody fragments, e.g., heavy chain variable (V) antibodies) specific for GPRC5D. H ) regions, single domain antibody fragments, and single chain fragments, e.g., scFvs). Also provided are cells (e.g., engineered or recombinant cells) that express such GPRC5D-binding receptors (e.g., anti-GPRC5D CARs) and / or contain nucleic acids encoding such receptors, as well as compositions and articles of manufacture and therapeutic doses comprising such cells.

[0192] Adoptive T cell therapy, such as CAR-T cell therapy, has shown promise for the treatment of multiple myeloma, with clinical efforts primarily focused on targeting B-cell maturation antigen (BCMA). However, although BCMA is expressed on many malignant plasma cells, expression levels can vary in some cases. In some cases, heterogeneity in target antigen expression can lead to variable or inconsistent responses. In some cases, BCMA expression on the cell surface has also been observed to change over time due to gamma secretase-mediated cleavage of the extracellular domain. Similar to observations with CD19 and CD22 CAR antigens, downregulation of BCMA antigen has been reported in relapsed multiple myeloma (MM) patients after BCMA-targeted T cell therapy (Brudno et al. (2018) J. Clin. Oncol., JCO2018778084,; Cohen et al. (2017) Blood 130:505). Additionally, in some situations, recombinant receptors may exhibit antigen-independent activity or signaling (also known as "tonic signaling"), which may result in undesirable effects due to, for example, increased differentiation and / or exhaustion of T cells expressing the recombinant receptor. In some aspects, such activity may limit the activity, efficacy, or potency of the T cells. In some cases, upon engineering and ex vivo expansion of cells to express the recombinant receptor, the cells may exhibit an exhaustive phenotype due to tonic signaling through the recombinant receptor. In some cases, alternative or additional MM-targeted T cell therapy approaches are needed.

[0193] An embodiment provided relates to GPRC5D as a target for CAR T cells in multiple myeloma. GPRC5D (Uniprot Acc. No. Q9NZD1, e.g., as shown in SEQ ID NO:49) is a G protein-coupled receptor class C group 5 member D belonging to the RAIG (retinoic acid-inducible gene-1) family. It is a 39 kDa G protein-coupled receptor with seven transmembrane helices, and two isoforms have been reported, with the difference between the isoforms residing in the intracellular C-terminus of the protein. Results herein demonstrate that GPRC5D is expressed at high levels in multiple myeloma and at low levels overall in most normal tissues.

[0194] The observations herein demonstrate protein expression of GPRC5D on multiple myeloma cells, supporting its role as a viable target for CAR T cells for the treatment of MM, based on, for example, assessment of potential on-target / off-tumor toxicity. Furthermore, some chimeric antigen receptors provided exhibit low tonic signaling, thereby minimizing the potential for antigen-independent (tonic) signaling. In particular, the anti-GPRC5D CARs provided herein include CARs with high antigen-dependent activation and minimal tonic signaling. In particular, certain constructs, such as those in which the variable heavy chain (VH) and variable light chain (VL) in the extracellular portion of the antibody fragment of the CAR are oriented in a particular direction and / or contain a particular length of spacer, have been found to exhibit advantageous properties, such as high antigen-dependent activation and low tonic signaling, compared to alternative anti-GPRC5D CAR formats, such as those with shorter spacers.

[0195] In some embodiments, the spacer is generally an amino acid sequence located between the extracellular antigen-binding domain and the transmembrane domain of the CAR, linking both domains. In particular embodiments of anti-GPRC5D CARs, the spacer is a portion of an immunoglobulin, e.g., from IgG4 or IgG2, such as a portion including the hinge domain, CH2 domain, and CH3 domain. Such spacers include portions of human immunoglobulins or modified forms thereof, e.g., those having a length of more than 125 amino acids, e.g., more than 150 amino acids, more than 180 amino acids, more than 200 amino acids, or more than 200 amino acids. In some embodiments, the immunoglobulin spacer is a hybrid or chimeric spacer and / or has been modified, e.g., to reduce or inhibit glycosylation. In some embodiments, the provided anti-GPRC5D CAR comprises an IgG4 / IgG2 hinge-IgG4 / IgG2 CH2-IgG4 CH3 immunoglobulin hybrid / modified spacer, e.g., as shown in SEQ ID NO: 17.

[0196] In some embodiments, some CARs provided herein are encoded by polynucleotides that have been optimized (e.g., codon usage) or contain specific features designed to reduce RNA heterogeneity and / or improve expression (e.g., surface expression) of the encoded receptor (e.g., to increase expression or make expression more consistent between cell product lots). In some embodiments, the polynucleotide encoding the cell surface protein that binds to GPRC5D has been modified compared to a reference polynucleotide, e.g., to remove cryptic or hidden splice sites, thereby reducing RNA heterogeneity. In some embodiments, the polynucleotide encoding the cell surface protein that binds to GPRC5D has been codon-optimized for expression in, e.g., mammalian cells, e.g., human cells, e.g., human T cells. In some aspects, the modified polynucleotide results in improved, e.g., increased, or more uniform or consistent expression levels, e.g., surface expression levels, when expressed in a cell. Such polynucleotides can be used in constructs to generate modified cells that express the cell surface proteins that bind to GPRC5D that they encode. Thus, also provided are cells that express recombinant receptors encoded by the polynucleotides provided herein and their use in adoptive cell therapy, for example, in the treatment of diseases and disorders associated with GPRC5D expression, such as multiple myeloma.

[0197] Provided is a monotherapy approach that utilizes anti-GPRC5D CARs expressed on primary autologous T cells for use as therapeutic agents against plasma cells of multiple myeloma. In some embodiments, the monotherapy approach may be desirable for subjects who are known to have, suspected to have, or selected as having MM plasma cells with low or no BCMA expression, and / or who have relapsed after remission, are refractory to anti-BCMA CARs, have failed treatment with anti-BCMA CARs, or are intolerant to treatment with anti-BCMA CARs.

[0198] Also provided herein is a multi-targeting strategy that targets a first antigen and a second antigen associated with a specific disease or condition (e.g., multiple myeloma). In some embodiments, multiple recombinant receptors that specifically bind to or specifically target different antigens are encoded by the same polynucleotide construct or are included in the same cell, composition, and method provided herein. In some embodiments, the multiple antigens (e.g., the first antigen and the second antigen) are expressed or suspected to be expressed on the surface of a target cell, tissue, or disease or condition, such as a cancer cell. In some aspects, the cell, tissue, disease, or condition is multiple myeloma or multiple myeloma cells.

[0199] Also provided herein is a two-drug combination therapy targeting approach, for example, using anti-GPRC5D CAR-expressing cells in combination with anti-BMCA CAR-expressing cells as a therapeutic agent for MM plasma cells.In some aspects, dual targeting approach can be advantageous for solving the limitations associated with the diverse expression of BCMA and / or GPRC5D in MM plasma cells.GPRC5D and BCMA are observed to be expressed, for example, diversely expressed, in certain diseases and pathologies, such as malignant tumors, or in tissues or their cells, for example, in malignant plasma cells derived from relapsed myeloma patients or newly diagnosed myeloma patients, and are rarely expressed in normal tissues.Due to the role of GPRC5D and BCMA in various diseases and pathologies, such as cancer, both GPRC5D and BCMA are therapeutic targets.

[0200] In some cases, simultaneously targeting both antigens provided herein may improve the depth and duration of response in patients, as well as minimize relapse due to antigen escape. The mechanism of resistance to CAR T cell therapy may be loss or downregulation ("escape") of the target antigen, as evidenced by data from CAR T cell trials in B-cell malignancies (Robbie G. Majzner and Crystal L. Mackall, Cancer Discov August 22, 2018; DOI 10.1158 / 2159-8290. CD-18-0442). Such combination or dual targeting strategies may result in synergistic or improved tumor responses based on targeting two antigens compared with monotherapy approaches that target only one antigen. Indeed, studies herein have demonstrated that BCMA expression and GPRC5D expression are independent of each other. A dual targeting approach may be advantageous for addressing potential antigen loss issues and / or maximizing antigen targeting in MM. The observations herein demonstrate protein expression of GPRC5D, BCMA, or both, on multiple myeloma cells and support that both antigens are viable targets of CAR T cells for the treatment of MM, based, for example, on assessment of potential on-target / off-tumor toxicities.

[0201] Among the embodiments provided are approaches useful for treating diseases and conditions and / or for targeting such cell types, e.g., nucleic acid molecules encoding receptors that bind GPRC5D and receptors that bind BCMA, e.g., chimeric antigen receptors (CARs), as well as the encoded receptors, e.g., the encoded CARs, and compositions and articles of manufacture comprising the same. The receptors are generally directed against antibodies (antigen-binding antibody fragments, e.g., heavy chain variable (V)) specific for GPRC5D or BCMA. H) regions, single-domain antibody fragments, and single-chain fragments, such as single-chain variable fragments (scFvs). Also provided are cells (e.g., engineered or recombinant cells) that express such GPRC5D-binding receptors (e.g., anti-GPRC5D CARs) and BCMA-binding receptors (e.g., anti-BCMA CARs) and / or contain nucleic acids encoding such receptors, as well as compositions, articles of manufacture, and therapeutic doses comprising such cells. Among the embodiments provided are polynucleotides that are bicistronic for the expression of multiple CARs (e.g., anti-GPRC5D CARs and anti-BCMA CARs). Observations herein demonstrate that the expression of multiple CARs (e.g., anti-GPRC5D CARs and anti-BCMA CARs) in cells can be improved by codon divergence in the polynucleotide sequence encoding one or more of the CARs. It has been found that codon divergence in polynucleotide constructs encoding two CARs improves expression of the 3' (or C-terminal) CAR-encoding nucleotide sequence relative to the nucleotide sequence encoding the other CAR.

[0202] Furthermore, it has been found that the provided CARs containing a spacer of a certain length exhibit advantageous properties, such as high antigen-dependent activation and low tonic signaling, compared to alternative anti-GPRC5D or anti-BCMA CAR formats, such as those with shorter spacers. In some embodiments, the spacer component of the CAR is generally an amino acid sequence located between the extracellular antigen-binding domain and the transmembrane domain of the CAR, linking both domains. In particular embodiments of anti-GPRC5D or anti-BCMA CARs, the spacer is a portion of an immunoglobulin, for example, an IgG4- or IgG2-derived portion, such as a portion comprising the hinge domain, CH2 domain, and CH3 domain. Among such spacers are portions of human immunoglobulins or modified forms thereof, such as those having a length of more than 125 amino acids, for example, more than 150 amino acids, more than 180 amino acids, more than 200 amino acids, or more than 200 amino acids. In some embodiments, the immunoglobulin spacer is a hybrid or chimeric spacer, and / or is modified, for example, to reduce or inhibit glycosylation. In some embodiments, the provided anti-GPRC5D CAR or anti-BCMA CAR comprises an IgG4 / IgG2 hinge-IgG4 / IgG2 CH2-IgG4 CH3 immunoglobulin hybrid / modified spacer, for example, as shown in SEQ ID NO:17. In particular embodiments, the polynucleotide encoding the CAR comprises a spacer region that has been modified to remove splice sites, for example, cryptic splice sites and / or acceptor sites. Exemplary nucleotides encoding the spacer are described. In some embodiments, the coding sequence of the spacer comprises the nucleic acid sequence shown in SEQ ID NO:48 (also shown in SEQ ID NO:74). In some embodiments, the provided CAR exhibits reduced RNA heterogeneity when expressed in cells (e.g., T cells). In some embodiments, the provided polynucleotide encoding the CAR can also be codon-optimized to further improve expression.

[0203] All publications referenced in this application, including patent documents, scientific papers, and databases, are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication were individually incorporated by reference. To the extent that a definition set forth herein conflicts or is otherwise inconsistent with a definition set forth in a patent, patent application, published patent application, or other publication incorporated herein by reference, the definition set forth herein shall take precedence over the definition incorporated herein by reference.

[0204] The section headings used herein are for organizational purposes only and should not be construed as limiting the subject matter described.

[0205] I. Receptors that bind to GPRC5D and encoding polynucleotides In some aspects, GPRC5D-binding agents, such as recombinant receptors or chimeric antigen receptors that bind to GPRC5D molecules, and cell surface proteins that bind to GPRC5D, such as polynucleotides encoding recombinant receptors (e.g., CARs) that bind to GPRC5D, and cells that express such receptors, are provided. Cell surface proteins that bind to GPRC5D generally include antibodies (e.g., antigen-binding antibody fragments) and / or other binding peptides that specifically bind to GPRC5D, such as GPRC5D proteins, for example, human GPRC5D proteins. In some aspects, the agents bind to the extracellular portion of GPRC5D.

[0206] Among the provided polynucleotides are those that encode recombinant receptors, e.g., antigen receptors, that specifically bind to GPRC5D. In some aspects, the encoded receptors, e.g., those comprising GPRC5D-binding polypeptides, as well as compositions and articles of manufacture and uses of the receptors, are also provided.

[0207] GPRC5D-binding polypeptides include antibodies, such as single-chain antibodies (e.g., antigen-binding antibody fragments), or portions thereof. In some examples, the recombinant receptor comprises a chimeric antigen receptor, such as an anti-GPRC5D antibody or an antigen-binding fragment thereof. The provided polynucleotides can be incorporated into a construct, such as a deoxyribonucleic acid (DNA) construct or an RNA construct, such as one that can be introduced into a cell for expression of the encoded recombinant GPRC5D-binding receptor.

[0208] 1. Encoded recombinant GPRC5D-binding receptor components The provided GPRC5D-binding receptors generally comprise an extracellular binding molecule and an intracellular signaling domain. Among the provided receptors are polypeptides comprising antibodies, such as recombinant cell surface receptors comprising anti-GPRC5D antibodies. Such receptors include chimeric antigen receptors comprising such antibodies.

[0209] Among the recombinant receptors provided are antigen receptors comprising GPRC5D-binding fragments. Such recombinant receptors include antigen receptors that specifically bind to GPRC5D, such as anti-GPRC5D antibodies, including antigen receptors comprising GPRC5D antigen-binding fragments. Among such antigen receptors are functional non-TCR antigen receptors, such as chimeric antigen receptors (CARs). Also provided are cells expressing the recombinant receptors and their use in adoptive cell therapy, for example, in the treatment of diseases and disorders associated with GPRC5D expression, such as multiple myeloma.

[0210] a. Extracellular antigen-binding domain Chimeric receptors include chimeric antigen receptors (CARs). Chimeric receptors such as CARs generally comprise an extracellular antigen-binding domain that includes, is, or comprises an anti-GPRC5D antibody. Thus, chimeric receptors, such as CARs, typically comprise one or more GPRC5D-binding molecules, such as one or more antigen-binding fragments, domains, or portions thereof, or one or more antibody variable regions, and / or antibody molecules, for example, as described herein, in their extracellular portion.

[0211] The term "antibody" is used herein in the broadest sense and includes polyclonal and monoclonal antibodies, including intact antibodies and functional (antigen-binding) antibody fragments, such as fragment antigen binding (Fab) fragments, F(ab')2 fragments, Fab' fragments, Fv fragments, recombinant IgG (rIgG) fragments, heavy chain variable (V) fragments capable of specifically binding to an antigen, and antibodies containing ... H ) regions, single-chain antibody fragments, such as single-chain variable fragments (scFvs), and single-domain antibody (e.g., sdAb, sdFv, nanobody) fragments. The term encompasses genetically engineered and / or otherwise modified forms of immunoglobulins, such as intrabodies, peptibodies, chimeric antibodies, fully human antibodies, humanized antibodies, as well as heteroconjugate antibodies, multispecific, e.g., bispecific or trispecific, antibodies, diabodies, triabodies, and tetrabodies, tandem di-scFvs, and tandem tri-scFvs. Unless otherwise specified, the term "antibody" is understood to encompass functional antibody fragments thereof, also referred to herein as "antigen-binding fragments." The term also encompasses intact or full-length antibodies, e.g., antibodies of any class or subclass, e.g., IgG and its subclasses, IgM, IgE, IgA, and IgD.

[0212] The terms "complementarity determining region" and "CDR," synonymous with "hypervariable region" or "HVR," are known in the art to refer to non-contiguous amino acid sequences within the variable region of an antibody that confer antigen specificity and / or binding affinity. Generally, there are three CDRs (CDR-H1, CDR-H2, CDR-H3) in each heavy chain variable region and three CDRs (CDR-L1, CDR-L2, CDR-L3) in each light chain variable region. The terms "framework region" and "FR" are known in the art to refer to the non-CDR portions of the heavy and light chain variable regions. Generally, there are four FRs (FR-H1, FR-H2, FR-H3, and FR-H4) in each full-length heavy chain variable region and four FRs (FR-L1, FR-L2, FR-L3, and FR-L4) in each full-length light chain variable region.

[0213] The exact amino acid sequence boundaries of a given CDR or FR can be determined according to any of several well-known schemes, e.g., Kabat et al., (1991), "Sequences of Proteins of Immunological Interest," 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD ("Kabat" numbering scheme); Al-Lazikani et al., (1997) JMB 273, 927-948 ("Chothia" numbering scheme); MacCallum et al., J. Mol. Biol. 262:732-745 (1996), "Antibody-antigen interactions: Contact analysis and binding site topography," J. Mol. Biol. 262, 732-745." ("Contact" numbering scheme); Lefranc MP et al., "IMGT unique numbering for immunoglobulin and T cell receptor variable domains and Ig superfamily V-like domains," Dev Comp Immunol, 2003 Jan; 27 (1): 55-77 ("IMGT" numbering scheme); Honegger A and Plueckthun A, "Yet another numbering scheme for immunoglobulin variable domains: an automatic modeling and analysis tool," J Mol Biol, 2001 Jun 8; 309 (3): 657-70, ("Aho" numbering scheme); and Martin et al., "Modeling antibody hypervariable loops: a combined algorithm," PNAS, 1989, 86 (23): 9268-9272, ("AbM" numbering scheme).

[0214] The boundaries of a given CDR or FR may vary depending on the scheme used for identification. For example, the Kabat scheme is based on structural alignment, while the Chothia scheme is based on structural information. Both the Kabat and Chothia numbering schemes are based on the full-length sequences of the most common antibody regions; some antibodies have deletions and insertions addressed by insertion letters, e.g., "30a." Certain insertions and deletions ("indels") are located at different positions in the two schemes, resulting in different numbering. The Contact scheme is based on analysis of complex crystal structures and is similar in many ways to the Chothia numbering scheme. The AbM scheme is a compromise between the Kabat and Chothia definitions, based on those used by Oxford Molecular's AbM antibody modeling software.

[0215] Table 1 below lists exemplary boundary locations for CDR-L1, CDR-L2, CDR-L3, and CDR-H1, CDR-H2, and CDR-H3, as identified by the Kabat, Chothia, AbM, and Contact schemes, respectively. For CDR-H1, residue numbering is listed using both the Kabat and Chothia numbering schemes. FRs are located between the CDRs; e.g., FR-L1 precedes CDR-L1, FR-L2 is located between CDR-L1 and CDR-L2, FR-L3 is located between CDR-L2 and CDR-L3, etc. Note that because the Kabat numbering scheme shown places the insertion at H35A and H35B, the endpoint of the Chothia CDR-H1 loop when numbered using the Kabat numbering scheme shown varies between H32 and H34 depending on the length of the loop.

[0216] Table 1. CDR boundaries according to various numbering schemes TIFF2025183309000020.tif471611 - Kabat et al., (1991), "Sequences of Proteins of Immunological Interest," 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD 2 - Al-Lazikani et al., (1997) JMB 273, 927-948

[0217] Thus, unless otherwise specified, the "CDRs" or "complementary determining regions" of a given antibody or region thereof, e.g., its variable region, or the individual CDRs designated (e.g., CDR-H1, CDR-H2, CDR-H3) should be understood to encompass a (or specific) complementarity determining region as defined by any of the schemes described above or other known schemes. For example, if a particular CDR (e.g., CDR-H3) is associated with a given V H or V L When a variable region is described as comprising the amino acid sequence of a corresponding CDR within its amino acid sequence, it is understood that such CDR has the sequence of the corresponding CDR (e.g., CDR-H3) within the variable region defined by any of the above-mentioned schemes or other known schemes. In some embodiments, specific CDR sequences are specified. While exemplary CDR sequences of the provided antibodies are described using various numbering schemes, it is understood that the provided antibodies may comprise CDRs described by any of the other numbering schemes described above or other numbering schemes known to those of skill in the art.

[0218] Similarly, unless otherwise specified, a given antibody or region thereof, such as the FRs of its variable region or designated individual FRs (e.g., FR-H1, FR-H2, FR-H3, FR-H4), is understood to encompass a (or specific) framework region defined by any known scheme. In some cases, a scheme for identifying a particular CDR, FR, or multiple FRs or CDRs is specified, such as CDRs defined by the Kabat, Chothia, AbM, or Contact method or other known schemes. In other cases, the specific amino acid sequence of a CDR or FR is provided.

[0219] The term "variable region" or "variable domain" refers to the domain of an antibody heavy or light chain that is involved in binding the antibody to an antigen. The variable regions of the heavy and light chains of a native antibody (V H and V L ) have generally similar structures, with each domain containing four conserved framework regions (FRs) and three CDRs. (See, e.g., Kindt et al., Kuby Immunology, 6th ed., W.H. Freeman and Co., page 91 (2007). A single V H Domain or V L The V domain may be sufficient to confer specificity to antigen binding. Furthermore, an antibody that binds to a particular antigen may have a V domain derived from the antibody that binds to that antigen. H Domain or V L The V domains are isolated and each has a complementary V H Domain or V L Libraries of domains can be screened. (See, e.g., Portolano et al., J. Immunol. 150:880-887 (1993); Clarkson et al., Nature 352:624-628 (1991).)

[0220] Among the antibodies contained in the provided CARs are antibody fragments. An "antibody fragment" or "antigen-binding fragment" refers to a molecule other than an intact antibody that contains a portion of the intact antibody that binds to the antigen to which the intact antibody binds. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2; diabodies; linear antibodies; heavy chain variable (V); and H ) regions, single chain antibody molecules, such as scFv and V H In some embodiments, the antigen-binding domain in the provided CAR is a heavy chain variable (V H ) and light chain variable (V L In certain embodiments, the antibody is or comprises an antibody fragment comprising a heavy chain variable (V H ) region and / or light chain variable (V L ) region, such as a single-chain antibody fragment, e.g., scFv.

[0221] A single domain antibody (sdAb) is an antibody fragment that contains all or a portion of the heavy chain variable region or all or a portion of the light chain variable region of an antibody. In certain embodiments, the single domain antibody is a human single domain antibody.

[0222] Antibody fragments can be produced by various techniques, including, but not limited to, proteolytic digestion of intact antibodies and production by recombinant host cells. In some embodiments, the antibody is a recombinantly produced fragment, e.g., a fragment containing a non-naturally occurring structure, e.g., two or more antibody regions or chains linked by a synthetic linker (e.g., a peptide linker), and / or a fragment containing a structure that cannot be produced by enzymatic digestion of a naturally occurring intact antibody. In some aspects, the antibody fragment is an scFv.

[0223] A "humanized" antibody is an antibody in which all or substantially all CDR amino acid residues are derived from non-human CDRs and all or substantially all FR amino acid residues are derived from human FRs. A humanized antibody may optionally contain at least a portion of an antibody constant region derived from a human antibody. A "humanized form" of a non-human antibody refers to a variant of a non-human antibody that has been humanized, typically to reduce immunogenicity to humans, but that retains the specificity and affinity of the non-human parent antibody. In some embodiments, some FR residues in a humanized antibody are substituted with corresponding residues from a non-human antibody (e.g., the antibody from which the CDR residues are derived), e.g., to restore or improve antibody specificity or affinity.

[0224] Among the anti-GPRC5D antibodies provided are human antibodies. A "human antibody" is an antibody having an amino acid sequence corresponding to that of an antibody produced by a human or human cell, or by a non-human source in which a human antibody repertoire or other human antibody coding sequence is utilized, such as a human antibody library. The term excludes humanized forms of non-human antibodies that contain non-human antigen-binding regions, such as those in which all or substantially all CDRs are non-human CDRs. The term includes antigen-binding fragments of human antibodies.

[0225] Human antibodies can be prepared by administering immunogens to transgenic animals that have been modified to produce intact human antibodies or intact antibodies with human variable regions in response to antigenic challenge. Such animals typically contain all or part of a human immunoglobulin locus that replaces the endogenous immunoglobulin locus or is present extrachromosomally or randomly integrated into the animal's chromosomes. In such transgenic animals, the endogenous immunoglobulin locus is generally inactivated. Human antibodies can also be derived from human antibody libraries, such as phage display and cell-free libraries, that contain antibody coding sequences from the human repertoire.

[0226] Among the antibodies contained in the provided CAR are monoclonal antibodies (including monoclonal antibody fragments). As used herein, the term "monoclonal antibody" refers to an antibody obtained from or within a substantially homogeneous antibody population, i.e., the individual antibodies comprising the population are identical except for possible variants that include naturally occurring mutations or that arise during the production of the monoclonal antibody preparation, and such variants are generally present in minor amounts. In contrast to polyclonal antibody preparations, which typically contain various antibodies directed against different epitopes, each monoclonal antibody in a monoclonal antibody preparation is directed against a single epitope on an antigen. This term should not be construed as requiring that the antibody be produced by any particular method. Monoclonal antibodies can be produced by a variety of techniques, including, but not limited to, hybridoma generation, recombinant DNA technology, phage display, and other antibody display methods.

[0227] In some embodiments, the CAR comprises the GPRC5D-binding portion of an antibody molecule, e.g., the variable heavy chain (V H ) region and / or light chain variable (V L ) region, e.g., an scFv antibody fragment. In some embodiments, the provided CARs that bind to GPRC5D comprise an antibody, e.g., an anti-GPRC5D antibody or antigen-binding fragment thereof, that confers the GPRC5D-binding properties of the provided CAR. In some embodiments, the antibody or antigen-binding domain can be any reported anti-GPRC5D antibody or can be derived from any reported anti-GPRC5D antibody (see, e.g., WO2016 / 090312, WO2016 / 090329, WO2018 / 017786). Any such anti-GPRC5D antibody or antigen-binding fragment can be used in the provided CARs. In some embodiments, the anti-GPRC5D CARs comprise a heavy chain variable (V) region derived from an antibody described in WO2016 / 090312, WO2016 / 090329, or WO2018 / 017786. H ) and / or light chain variable (V L) region.

[0228] In some embodiments, the antibody, e.g., anti-GPRC5D antibody or antigen-binding fragment, comprises a heavy chain variable and / or a light chain variable (V H or V L In some embodiments, the anti-GPRC5D antibody, e.g., antigen-binding fragment, comprises a V domain sequence comprising CDR-H1, CDR-H2, and / or CDR-H3 as described above. H In some embodiments, the anti-GPRC5D antibody, e.g., antigen-binding fragment, comprises a V domain sequence comprising CDR-L1, CDR-L2, and / or CDR-L3 as described above, or a sufficient antigen-binding portion thereof. L In some embodiments, the anti-GPRC5D antibody, e.g., antigen-binding fragment, comprises a V domain sequence or a sufficient antigen-binding portion thereof, including CDR-H1, CDR-H2, and / or CDR-H3 as described. H V comprising the V region sequence and including CDR-L1, CDR-L2 and / or CDR-L3 as described L Also included are antibodies that have a sequence that is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% identical to such a sequence.

[0229] In some embodiments, the antibody or antibody fragment in the provided CAR comprises the V of any antibody or antibody-binding fragment described in any of WO2016 / 090312, WO2016 / 090329, and WO2018 / 017786. H It has an area.

[0230] In some embodiments, the CAR comprises a heavy chain variable (V) having an amino acid sequence selected from any one of SEQ ID NOs: 21, 23, 25, 27, 29, 31, or 33. H ) region, or any one of SEQ ID NOs: 21, 23, 25, 27, 29, 31, or 33. H V having an amino acid sequence having at least 90% or at least about 90%, at least 91% or at least about 91%, at least 92% or at least about 92%, at least 93% or at least about 93%, at least 94% or at least about 94%, at least 95% or at least about 95%, at least 96% or at least about 96%, at least 97% or at least about 97%, at least 98% or at least about 98%, or at least 99% or at least about 99% sequence identity to the amino acids in the region. H have a region or the V H The present invention includes an antibody or antigen-binding fragment thereof comprising CDR-H1, CDR-H2, and / or CDR-H3 present in the sequence.

[0231] In some embodiments, the V of the antibody or antigen-binding fragment thereof H The region comprises CDR-H1, CDR-H2, and / or CDR-H3 according to the Kabat numbering. H The region comprises CDR-H1, CDR-H2, and / or CDR-H3 according to Chothia numbering. H The regions include CDR-H1, CDR-H2, and / or CDR-H3 according to AbM numbering.

[0232] In some embodiments, the CAR comprises: (a) a CDR-H1 comprising an amino acid sequence selected from SEQ ID NOs: 75, 78, 80, 82, 90, 93, 95, 97, 105, 108, 110, 112, 120, 123, 125, 127, 135, 138, 140, 142, 152, 162, 165, 167, and 169; (b) a CDR-H2 comprising an amino acid sequence selected from SEQ ID NOs: 76, 79, 81, 83, 91, 94, 96, 98, 106, 109, 111, 113, 121, 124, 126, 128, 136, 139, 141, 143, 150, 153, 154, 155, 163, 166, 168, and 170; and (c) a CDR-H3 comprising an amino acid sequence selected from SEQ ID NOs: 76, 79, 81, 83, 91, 94, 96, 98, 106, 109, 111, 113, 121, 124, 126, 128, 136, 139, 141, 143, 150, 153, 154, 155, 163, 166, 168, and 170. A heavy chain variable (V) comprising a CDR-H3 comprising an amino acid sequence selected from NOs: 77, 84, 92, 99, 107, 114, 122, 129, 137, 144, 151, 156, 164, and 171. H ) region or an antigen-binding fragment thereof.

[0233] In some embodiments, the antibodies or antigen-binding fragments thereof are selected from the group consisting of SEQ ID NOs: 75, 76, and 77, respectively; SEQ ID NOs: 78, 79, and 77, respectively; SEQ ID NOs: 80, 81, and 77, respectively; SEQ ID NOs: 82, 83, and 84, respectively; SEQ ID NOs: 90, 91, and 92, respectively; SEQ ID NOs: 93, 94, and 92, respectively; SEQ ID NOs: 95, 96, and 92, respectively; SEQ ID NOs: 97, 98, and 99, respectively; SEQ ID NOs: 105, 106, and 107, respectively; SEQ ID NOs: 108, 109, and 107, respectively; SEQ ID NOs: 110, 111, and 107, respectively; SEQ ID NOs: 112, 113, and 114, respectively; SEQ ID NOs: 120, 121, and 122, respectively; SEQ ID NOs: 123, 124, and 122, respectively; and SEQ ID NOs: 125, 126, and 127, respectively. SEQ ID NOs: 125, 126 and 122, respectively; SEQ ID NOs: 127, 128 and 129, respectively; SEQ ID NOs: 135, 136 and 137, respectively; SEQ ID NOs: 138, 139 and 137, respectively; SEQ ID NOs: 140, 141 and 137, respectively; SEQ ID NOs: 142, 143 and 144, respectively; SEQ ID NOs: 135, 150 and 151, respectively; SEQ ID NOs: 152, 153 and 151, respectively; SEQ ID NOs: 140, 154 and 151, respectively; SEQ ID NOs: 142, 155 and 156, respectively; SEQ ID NOs: 162, 163 and 164, respectively; SEQ ID NOs: 165, 166 and 164, respectively; SEQ ID NOs: 167, 168 and 164, respectively; SEQ ID NOs: 169, 161 and 162 ... V including CDR-H1, CDR-H2, and CDR-H3 containing the amino acid sequences of NO:169, 170, and 171 H Includes the area.

[0234] In some embodiments, the antibodies or antigen-binding fragments thereof are selected from the group consisting of SEQ ID NOs: 75, 76, and 77, respectively; SEQ ID NOs: 78, 79, and 77, respectively; SEQ ID NOs: 80, 81, and 77, respectively; SEQ ID NOs: 82, 83, and 84, respectively; SEQ ID NOs: 90, 91, and 92, respectively; SEQ ID NOs: 93, 94, and 92, respectively; SEQ ID NOs: 95, 96, and 92, respectively; SEQ ID NOs: 97, 98, and 99, respectively; SEQ ID NOs: 105, 106, and 107, respectively; SEQ ID NOs: 108, 109, and 107, respectively; SEQ ID NOs: 110, 111, and 107, respectively; SEQ ID NOs: 112, 113, and 114, respectively; SEQ ID NOs: 120, 121, and 122, respectively; SEQ ID NOs: 123, 124, and 122, respectively; and SEQ ID NOs: 125, 126, and 127, respectively. SEQ ID NOs: 125, 126 and 122, respectively; SEQ ID NOs: 127, 128 and 129, respectively; SEQ ID NOs: 135, 136 and 137, respectively; SEQ ID NOs: 138, 139 and 137, respectively; SEQ ID NOs: 140, 141 and 137, respectively; SEQ ID NOs: 142, 143 and 144, respectively; SEQ ID NOs: 135, 150 and 151, respectively; SEQ ID NOs: 152, 153 and 151, respectively; SEQ ID NOs: 140, 154 and 151, respectively; SEQ ID NOs: 142, 155 and 156, respectively; SEQ ID NOs: 162, 163 and 164, respectively; SEQ ID NOs: 165, 166 and 164, respectively; SEQ ID NOs: 167, 168 and 164, respectively; SEQ ID NOs: 169, 161 and 162 ... V containing the amino acid sequence of NO:169, 170 and 171 H Includes the area.

[0235] In some embodiments, the antibody or antigen-binding fragment thereof is selected from the group consisting of V, VB, VC, VD, VE, VF, VF, VH ... HThe amino acid sequences of CDR-H1, CDR-H2, and CDR-H3 are contained within the region amino acid sequence.

[0236] In some embodiments of the antibodies or antigen-binding fragments thereof provided herein, V H The region includes any of the CDR-H1, CDR-H2, and CDR-H3 sequences described and is a V region as set forth in any one of SEQ ID NOs: 21, 23, 25, 27, 29, 31, or 33. H The regions comprise framework region 1 (FR1), FR2, FR3, and / or FR4 that have at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% or at least about 99% sequence identity to FR1, FR2, FR3, and / or FR4, respectively, contained within the region amino acid sequence.

[0237] In some embodiments, the antibody or antigen-binding fragment thereof comprises a V comprising the amino acid sequence set forth in any one of SEQ ID NOs: 21, 23, 25, 27, 29, 31, or 33. H Includes the area.

[0238] In some embodiments, V H The antibody or antibody fragment within the provided CAR (e.g., anti-GPRC5D CAR) comprising the region further comprises a light chain or a sufficient antigen-binding portion thereof. For example, in some embodiments, the antibody or antigen-binding fragment thereof comprises a V H Area and V L Area or V H Area and V L In such an embodiment, the VH The region array is any of the above V H In some such embodiments, the antibody is an antigen-binding fragment, such as a Fab or scFv. In some such embodiments, the antibody is a full-length antibody that also includes a constant region.

[0239] In some embodiments, the CARs provided herein comprise any of the V H For example, in some embodiments, the CAR comprises an antibody comprising a V H Region and light chain variable (V L ) area or V H Area and V L In such embodiments, the V H The region array is any of the above V H In some such embodiments, the antibody is an antigen-binding fragment, such as a Fab or scFv. In some such embodiments, the antibody is a full-length antibody that also includes a constant region.

[0240] In some embodiments, the antibody or antigen-binding fragment is a V antibody or VIII antibody described in any of WO2016 / 090312, WO2016 / 090329, and WO2018 / 017786. L It has an area.

[0241] In some embodiments, the CAR comprises a light chain variable (V) having an amino acid sequence selected from any one of SEQ ID NOs: 22, 24, 26, 28, 30, 32, 34, 63, 64, 65, 66, 67, 68, or 69. L ) region, or any one of SEQ ID NOs: 22, 24, 26, 28, 30, 32, 34, 63, 64, 65, 66, 67, 68, or 69. LV having an amino acid sequence having at least 90% or at least about 90%, at least 91% or at least about 91%, at least 92% or at least about 92%, at least 93% or at least about 93%, at least 94% or at least about 94%, at least 95% or at least about 95%, at least 96% or at least about 96%, at least 97% or at least about 97%, at least 98% or at least about 98%, or at least 99% or at least about 99% sequence identity to the amino acids in the region. L have a region or the V L In some embodiments, the CAR comprises an antibody or antigen-binding fragment thereof comprising a light chain variable (V) having an amino acid sequence selected from any one of SEQ ID NOs: 22, 24, 26, 28, 30, 32, or 34. L ) region, or any one of SEQ ID NOs: 22, 24, 26, 28, 30, 32, or 34 L V having an amino acid sequence having at least 90% or at least about 90%, at least 91% or at least about 91%, at least 92% or at least about 92%, at least 93% or at least about 93%, at least 94% or at least about 94%, at least 95% or at least about 95%, at least 96% or at least about 96%, at least 97% or at least about 97%, at least 98% or at least about 98%, or at least 99% or at least about 99% sequence identity to the amino acids in the region. L have a region or the V L In some embodiments, the CAR comprises an antibody or antigen-binding fragment thereof comprising a light chain variable (V) having an amino acid sequence selected from any one of SEQ ID NOs: 63, 64, 65, 66, 67, 68, or 69. L) region, or any one of SEQ ID NOs: 63, 64, 65, 66, 67, 68, or 69. L V having an amino acid sequence having at least 90% or at least about 90%, at least 91% or at least about 91%, at least 92% or at least about 92%, at least 93% or at least about 93%, at least 94% or at least about 94%, at least 95% or at least about 95%, at least 96% or at least about 96%, at least 97% or at least about 97%, at least 98% or at least about 98%, or at least 99% or at least about 99% sequence identity to the amino acids in the region. L have a region or the V L The present invention includes an antibody or antigen-binding fragment thereof, comprising CDR-L1, CDR-L2, and / or CDR-L3 present in the sequence.

[0242] In some embodiments, the V of the antibody or antigen-binding fragment thereof L The region comprises CDR-L1, CDR-L2, and / or CDR-L3 according to the Kabat numbering system. L The region comprises CDR-L1, CDR-L2, and / or CDR-L3 according to Chothia numbering. L The regions include CDR-L1, CDR-L2, and / or CDR-L3 according to AbM numbering.

[0243] In some embodiments, the CAR comprises a light chain variable (V) CDR-L1 comprising an amino acid sequence selected from SEQ ID NOs: 85, 88, 100, 103, 115, 118, 130, 133, 145, 148, 157, 160, 172, and 174; (b) a CDR-L2 comprising an amino acid sequence selected from SEQ ID NOs: 86, 89, 101, 104, 116, 119, 131, 134, 146, 149, 158, and 161; and (c) a CDR-L3 comprising an amino acid sequence selected from SEQ ID NOs: 87, 102, 117, 132, 147, 159, 173, and 175. L ) region.

[0244] In some embodiments, the antibodies or antigen-binding fragments thereof are selected from the group consisting of SEQ ID NOs: 85, 86, and 87, respectively; SEQ ID NOs: 88, 89, and 87, respectively; SEQ ID NOs: 100, 101, and 102, respectively; SEQ ID NOs: 103, 104, and 102, respectively; SEQ ID NOs: 115, 116, and 117, respectively; SEQ ID NOs: 118, 119, and 117, respectively; SEQ ID NOs: 130, 131, and 132, respectively; SEQ ID NOs: 133, 134, and 132, respectively; SEQ ID NOs: 145, 146, and 147, respectively; SEQ ID NOs: 148, 149, and 147, respectively; SEQ ID NOs: 157, 158, and 159, respectively; SEQ ID NOs: 160, 161, and 159, respectively; SEQ ID NOs: 172, 86, and 173, respectively; and SEQ ID NOs: 174, 175, and 176, respectively. SEQ ID NOs: 174, 89, and 175; V containing CDR-L1, CDR-L2, and CDR-L3 containing the amino acid sequences of SEQ ID NOs: 174, 89, and 297, respectively. L Includes the area.

[0245] In some embodiments, the antibodies or antigen-binding fragments thereof are selected from the group consisting of SEQ ID NOs: 85, 86, and 87, respectively; SEQ ID NOs: 88, 89, and 87, respectively; SEQ ID NOs: 100, 101, and 102, respectively; SEQ ID NOs: 103, 104, and 102, respectively; SEQ ID NOs: 115, 116, and 117, respectively; SEQ ID NOs: 118, 119, and 117, respectively; SEQ ID NOs: 130, 131, and 132, respectively; SEQ ID NOs: 133, 134, and 132, respectively; SEQ ID NOs: 145, 146, and 147, respectively; SEQ ID NOs: 148, 149, and 147, respectively; SEQ ID NOs: 157, 158, and 159, respectively; SEQ ID NOs: 160, 161, and 159, respectively; SEQ ID NOs: 172, 86, and 173, respectively; and SEQ ID NOs: 174, 175, and 176, respectively. NOS: 174, 89, and 175; V containing the amino acid sequences of SEQ ID NOS: 174, 89, and 297, respectively. L Includes the area.

[0246] In some embodiments, the antibody or antigen-binding fragment thereof is selected from any one of SEQ ID NOs: 22, 24, 26, 28, 30, 32, 34, 63, 64, 65, 66, 67, 68, or 69. L In some embodiments, the antibody or antigen-binding fragment thereof comprises a CDR-L1, CDR-L2, and CDR-L3, each of which is contained within a V region amino acid sequence selected from any one of SEQ ID NOs: 22, 24, 26, 28, 30, 32, or 34. L In some embodiments, the antibody or antigen-binding fragment thereof comprises a CDR-L1, CDR-L2, and CDR-L3, each of which is contained within a V region amino acid sequence selected from any one of SEQ ID NOs: 63, 64, 65, 66, 67, 68, or 69. L The regions include CDR-L1, CDR-L2, and CDR-L3, each contained within an amino acid sequence.

[0247] As used herein, the CARs include those in which the antibody, e.g., anti-GPRC5D antibody, or antibody fragment in the provided CAR has at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to the amino acid sequence set forth in any of SEQ ID NOs: 21, 23, 25, 27, 29, 31, or 33. H and a V region amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to the amino acid sequence set forth in any one of SEQ ID NOs: 22, 24, 26, 28, 30, 32, 34, 63, 64, 65, 66, 67, 68, or 69. L and CAR, which includes the region.

[0248] In some embodiments, the V of the antibody or antigen-binding fragment thereof H The region is selected from any one of SEQ ID NOs: 21, 23, 25, 27, 29, 31, or 33. Ha CDR-H1 region comprising the amino acid sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, contained within the amino acid sequence of the CDR-H1 region; a V region selected from any one of SEQ ID NOs: 22, 24, 26, 28, 30, 32, 34, 63, 64, 65, 66, 67, 68, or 69; L The amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 are contained within the amino acid sequences of the CDR-L1, CDR-L2, and CDR-L3 regions, respectively.

[0249] In some embodiments, the V of the antibody or antigen-binding fragment thereof H The region comprises the amino acid sequence of SEQ ID NO: 21, 23, 25, 27, 29, 31, or 33, and is a V region of the antibody or antigen-binding fragment. L The region comprises amino acid sequence 22, 24, 26, 28, 30, 32, or 34. In some embodiments, the V H Area and V L The region comprises the amino acid sequence of SEQ ID NOs: 21 and 22, respectively; SEQ ID NOs: 23 and 24, respectively; SEQ ID NOs: 25 and 26, respectively; SEQ ID NOs: 27 and 28, respectively; SEQ ID NOs: 29 and 30, respectively; SEQ ID NOs: 31 and 32, respectively; or SEQ ID NOs: 33 and 34, respectively, or any of the above. H and V L The present invention also includes any antibody or antigen-binding fragment thereof having at least 90% sequence identity to the above, for example, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity thereto.

[0250] For example, the V of an antibody or antigen-binding fragment thereof provided by the present invention H Area and V LThe regions comprise an amino acid sequence selected from SEQ ID NOs:21 and 22; SEQ ID NOs:23 and 24; SEQ ID NOs:25 and 26; SEQ ID NOs:27 and 28; SEQ ID NOs:29 and 30; SEQ ID NOs:31 and 32; and SEQ ID NOs:33 and 34. In another example, the V region of an antibody or antigen-binding fragment thereof provided herein comprises an amino acid sequence selected from SEQ ID NOs:21 and 22; SEQ ID NOs:23 and 24; SEQ ID NOs:25 and 26; SEQ ID NOs:27 and 28; SEQ ID NOs:29 and 30; SEQ ID NOs:31 and 32; and SEQ ID NOs:33 and 34. H Area and V L The regions comprise amino acid sequences selected from SEQ ID NOs:21 and 63; SEQ ID NOs:23 and 64; SEQ ID NOs:25 and 65; SEQ ID NOs:27 and 66; SEQ ID NOs:29 and 67; SEQ ID NOs:31 and 68; and SEQ ID NOs:33 and 69, respectively.

[0251] In some embodiments, the antibody or antigen-binding fragment in the provided CAR is a single-chain antibody fragment, such as a single-chain variable fragment (scFv) or a diabody or a single-domain antibody (sdAb). H In some embodiments, the antibody or antigen-binding fragment is a single domain antibody comprising only the heavy chain variable (V H ) region and the light chain variable (V L In some embodiments, a single-chain antibody fragment (e.g., an scFv) comprises two antibody domains or regions, such as a heavy chain variable (V H ) region and the light chain variable (V L ) regions. The linker is typically a peptide linker, e.g., a flexible and / or soluble peptide linker. Some linkers are glycine / serine-rich and / or in some cases threonine-rich. In some embodiments, the linker further comprises charged residues, e.g., lysine and / or glutamic acid, which may improve solubility. In some embodiments, the linker further comprises one or more prolines.

[0252] Thus, the provided CARs typically comprise two antibody domains or regions, such as V H Area and V L Single-chain antibody fragments containing a linker connecting the domains include, for example, scFvs and diabodies, particularly anti-GPRC5D antibodies, including human single-chain antibody fragments. The linker is typically a peptide linker, for example, a flexible and / or soluble peptide linker, for example, a glycine / serine-rich one.

[0253] In some aspects, a glycine / serine (and / or threonine)-rich linker comprises at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of such amino acids. In some embodiments, it comprises at least 50%, or at least about 50%, at least 55%, or at least about 55%, at least 60%, at least 70%, or at least about 70%, or at least 75%, or at least about 75% glycine, serine, and / or threonine. In some embodiments, the linker is substantially entirely composed of glycine, serine, and / or threonine. Linkers are generally about 5 to about 50 amino acids in length, typically 10 or about 10 to 30 or about 30 amino acids in length, e.g., 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 amino acids in length, and in some examples, 10 to 25 amino acids in length. Exemplary linkers include linkers having various repeats of the sequence GGGGS (4GS; SEQ ID NO:50) or GGGS (3GS; SEQ ID NO:51), e.g., 2, 3, 4, or 5 repeats of such a sequence. Exemplary linkers include: TIFF2025183309000021.tif4128 or consisting of the sequence shown in the sequence. Further examples include those having or consisting of the sequence shown in TIFF2025183309000022.tif4128. Exemplary linkers include: Further examples include those having or consisting of the sequence shown in TIFF2025183309000023.tif4128. Exemplary linkers include: Examples include those having the sequence shown in TIFF2025183309000024.tif4128 or those consisting of said sequence.

[0254] Thus, in some embodiments, provided embodiments include single chain antibody fragments, e.g., scFvs, comprising one or more of the foregoing linkers, e.g., glycine / serine-rich linkers, e.g., linkers having repeats of GGGS (SEQ ID NO:51) or GGGGS (SEQ ID NO:50), e.g., linkers shown in SEQ ID NOs:47, 52, or 54.

[0255] In some embodiments, V H The area is V L In some embodiments, the V H The area is V L In certain embodiments, the fragment, e.g., the scFv, may have a V H A region or a portion thereof, followed by a linker, followed by V L In other embodiments, the fragment, e.g., an scFv, may include a V L A region or a portion thereof, followed by a linker, followed by V H A region or a portion thereof may be included.

[0256] In some aspects, the scFv provided herein comprises an amino acid sequence selected from any one of SEQ ID NOs: 1-14, or has an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to an amino acid sequence selected from any one of SEQ ID NOs: 1-14.

[0257] Among the anti-GPRC5D CARs provided, the antibody or antigen-binding fragment comprises a V that includes an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:21. Hand comprising the sequence set forth in SEQ ID NO:22, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:22. L In some embodiments, the provided CARs include CARs in which the antibody or antigen-binding fragment comprises a V region, wherein the antibody or antigen-binding fragment comprises an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:21. H and comprising the sequence set forth in SEQ ID NO:63, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:63. LIn some embodiments, the antibody or antigen-binding fragment of the CAR provided has a V region having CDRH1, CDRH2, and CDRH3 comprising the amino acid sequence of SEQ ID NOs: 75, 76, and 77, respectively. H and a V region having CDRL1, CDRL2, and CDRL3 comprising the amino acid sequences of SEQ ID NOs: 85, 86, and 87, respectively. L In some embodiments, the antibody or antigen-binding fragment of the provided CAR comprises a V region having a CDRH1, a CDRH2, and a CDRH3 comprising the amino acid sequences of SEQ ID NOs: 78, 79, and 77, respectively. H and V region including CDRL1, CDRL2, and CDRL3 containing the amino acid sequences of SEQ ID NOs: 85, 86, and 87, respectively. L In some embodiments, the antibody or antigen-binding fragment of the provided CAR comprises a V region having a CDRH1, a CDRH2, and a CDRH3 comprising the amino acid sequences of SEQ ID NOs: 80, 81, and 77, respectively. H and V region including CDRL1, CDRL2, and CDRL3 containing the amino acid sequences of SEQ ID NOs: 85, 86, and 87, respectively. L In some embodiments, the antibody or antigen-binding fragment of the provided CAR comprises a V region having a CDRH1, a CDRH2, and a CDRH3 comprising the amino acid sequences of SEQ ID NOs: 82, 83, and 84, respectively. H and V region including CDRL1, CDRL2, and CDRL3 containing the amino acid sequences of SEQ ID NOs: 88, 89, and 87, respectively. L In some embodiments, the V H The region comprises the sequence shown in SEQ ID NO:21, L The region comprises the sequence shown in SEQ ID NO:22. H The region comprises the sequence shown in SEQ ID NO:21, LThe region comprises the sequence set forth in SEQ ID NO:63. In some embodiments, the antibody or antigen-binding fragment is a single-chain antibody fragment, e.g., an scFv. In some embodiments, the scFv comprises the amino acid sequence set forth in SEQ ID NO:1, or an amino acid sequence at least or at least about 90%, at least 91%, or at least about 91%, at least 92%, or at least about 92%, at least 93%, or at least about 93%, at least 94%, or at least about 94%, at least 95%, or at least about 95%, at least 96%, or at least about 96%, at least 97%, or at least about 97%, at least 98%, or at least about 98%, or at least 99% or at least about 99% identical to SEQ ID NO:1. In some embodiments, the scFv is encoded by the nucleotide sequence set forth in SEQ ID NO:257, or a nucleotide sequence that is at least or at least about 90%, at least or at least about 91%, at least 92% or at least about 92%, at least 93% or at least about 93%, at least 94% or at least about 94%, at least 95% or at least about 95%, at least 96% or at least about 96%, at least 97% or at least about 97%, at least 98% or at least about 98%, or at least 99% or at least about 99% identical to SEQ ID NO:257. In some embodiments, the scFv comprises the amino acid sequence set forth in SEQ ID NO:2, or an amino acid sequence that is at least or at least about 90%, at least or at least about 91%, at least 92% or at least about 92%, at least 93% or at least about 93%, at least 94% or at least about 94%, at least 95% or at least about 95%, at least 96% or at least about 96%, at least 97% or at least about 97%, at least 98% or at least about 98%, or at least 99% or at least about 99% identical to SEQ ID NO:2.In some embodiments, the scFv is encoded by the nucleotide sequence set forth in SEQ ID NO:258, or a nucleotide sequence that is at least or at least about 90%, at least or at least about 91%, at least 92% or at least about 92%, at least 93% or at least about 93%, at least 94% or at least about 94%, at least 95% or at least about 95%, at least 96% or at least about 96%, at least 97% or at least about 97%, at least 98% or at least about 98%, or at least 99% or at least about 99% identical to SEQ ID NO:258.

[0258] Among the anti-GPRC5D CARs provided, the antibody or antigen-binding fragment comprises a V that includes an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 97%, at least 98%, or at least 98%, or at least 99% identity to SEQ ID NO:23. H and comprising the sequence set forth in SEQ ID NO:24, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:24. LIn some embodiments, the provided CARs include CARs in which the antibody or antigen-binding fragment comprises a V region, wherein the antibody or antigen-binding fragment comprises an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to the sequence set forth in SEQ ID NO:23. H and comprising the sequence set forth in SEQ ID NO:64, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:64. L In some embodiments, the antibody or antigen-binding fragment of the CAR provided has a V region having CDRH1, CDRH2, and CDRH3 comprising the amino acid sequences of SEQ ID NOs: 90, 91, and 92, respectively. H and a V region having CDRL1, CDRL2, and CDRL3 comprising the amino acid sequences of SEQ ID NOs: 100, 101, and 102, respectively. L In some embodiments, the antibody or antigen-binding fragment of the provided CAR comprises a V region having a CDRH1, a CDRH2, and a CDRH3 comprising the amino acid sequences of SEQ ID NOs: 93, 94, and 92, respectively. Hand a V region comprising CDRL1, CDRL2, and CDRL3 comprising the amino acid sequences of SEQ ID NOs: 100, 101, and 102, respectively. L In some embodiments, the antibody or antigen-binding fragment of the provided CAR comprises a V region having a CDRH1, a CDRH2, and a CDRH3 comprising the amino acid sequences of SEQ ID NOs: 95, 96, and 92, respectively. H and a V region comprising CDRL1, CDRL2, and CDRL3 comprising the amino acid sequences of SEQ ID NOs: 100, 101, and 102, respectively. L In some embodiments, the antibody or antigen-binding fragment of the provided CAR comprises a V region having a CDRH1, a CDRH2, and a CDRH3 comprising the amino acid sequences of SEQ ID NOs: 97, 98, and 99, respectively. H and V comprising CDRL1, CDRL2, and CDRL3 comprising the amino acid sequences of SEQ ID NOs: 103, 104, and 102, respectively. L In some embodiments, the V H The region comprises the sequence shown in SEQ ID NO:23, L The region comprises the sequence set forth in SEQ ID NO:24. H The region comprises the sequence shown in SEQ ID NO:23, LThe region comprises the sequence set forth in SEQ ID NO:64. In some embodiments, the antibody or antigen-binding fragment is a single-chain antibody fragment, e.g., an scFv. In some embodiments, the scFv comprises the amino acid sequence set forth in SEQ ID NO:3, or an amino acid sequence at least or at least about 90%, at least 91%, or at least about 91%, at least 92%, or at least about 92%, at least 93%, or at least about 93%, at least 94%, or at least about 94%, at least 95%, or at least about 95%, at least 96%, or at least about 96%, at least 97%, or at least about 97%, at least 98%, or at least about 98%, or at least 99% or at least about 99% identical to SEQ ID NO:3. In some embodiments, the scFv is encoded by the nucleotide sequence set forth in SEQ ID NO:259, or a nucleotide sequence that is at least or at least about 90%, at least 91% or at least about 91%, at least 92% or at least about 92%, at least 93% or at least about 93%, at least 94% or at least about 94%, at least 95% or at least about 95%, at least 96% or at least about 96%, at least 97% or at least about 97%, at least 98% or at least about 98%, or at least 99% or at least about 99% identical to SEQ ID NO:259. In some embodiments, the scFv comprises the amino acid sequence set forth in SEQ ID NO:4, or an amino acid sequence that is at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identical to SEQ ID NO:4.In some embodiments, the scFv is encoded by the nucleotide sequence set forth in SEQ ID NO:260, or a nucleotide sequence that is at least 90% or at least about 90%, at least 91% or at least about 91%, at least 92% or at least about 92%, at least 93% or at least about 93%, at least 94% or at least about 94%, at least 95% or at least about 95%, at least 96% or at least about 96%, at least 97% or at least about 97%, at least 98% or at least about 98%, or at least 99% or at least about 99% identical to SEQ ID NO:260.

[0259] Among the anti-GPRC5D CARs provided, the antibody or antigen-binding fragment comprises a V that includes an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 97%, at least 98%, or at least 98%, or at least 99% identity to SEQ ID NO:25. H and comprising the sequence set forth in SEQ ID NO:26, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:26. LIn some embodiments, the provided CARs include CARs in which the antibody or antigen-binding fragment comprises a V region, wherein the antibody or antigen-binding fragment comprises an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:25. H and comprising the sequence set forth in SEQ ID NO:65, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:65. L In some embodiments, the antibody or antigen-binding fragment of the CAR provided has a V region having CDRH1, CDRH2, and CDRH3 comprising the amino acid sequences of SEQ ID NOs: 105, 106, and 107, respectively. H and a V region having CDRL1, CDRL2, and CDRL3 comprising the amino acid sequences of SEQ ID NOs: 115, 116, and 117, respectively. L In some embodiments, the antibody or antigen-binding fragment of the provided CAR comprises a V region having a CDRH1, a CDRH2, and a CDRH3 comprising the amino acid sequences of SEQ ID NOs: 108, 109, and 107, respectively. Hand V comprising CDRL1, CDRL2, and CDRL3 comprising the amino acid sequences of SEQ ID NOs: 115, 116, and 117, respectively. L In some embodiments, the antibody or antigen-binding fragment of the provided CAR comprises a V region having a CDRH1, a CDRH2, and a CDRH3 comprising the amino acid sequences of SEQ ID NOs: 110, 111, and 107, respectively. H and V comprising CDRL1, CDRL2, and CDRL3 comprising the amino acid sequences of SEQ ID NOs: 115, 116, and 117, respectively. L In some embodiments, the antibody or antigen-binding fragment of the provided CAR comprises a V region having a CDRH1, a CDRH2, and a CDRH3 comprising the amino acid sequences of SEQ ID NOs: 112, 113, and 114, respectively. H and V comprising CDRL1, CDRL2, and CDRL3 comprising the amino acid sequences of SEQ ID NOs: 118, 119, and 117, respectively. L In some embodiments, the V H The region comprises the sequence shown in SEQ ID NO:25, L The region comprises the sequence set forth in SEQ ID NO: 26. H The region comprises the sequence shown in SEQ ID NO:25, LThe region comprises the sequence set forth in SEQ ID NO:65. In some embodiments, the antibody or antigen-binding fragment is a single-chain antibody fragment, e.g., an scFv. In some embodiments, the scFv comprises the amino acid sequence set forth in SEQ ID NO:5, or an amino acid sequence at least or at least about 90%, at least 91%, or at least about 91%, at least 92%, or at least about 92%, at least 93%, or at least about 93%, at least 94%, or at least about 94%, at least 95%, or at least about 95%, at least 96%, or at least about 96%, at least 97%, or at least about 97%, at least 98%, or at least about 98%, or at least 99% or at least about 99% identical to SEQ ID NO:5. In some embodiments, the scFv is encoded by the nucleotide sequence set forth in SEQ ID NO:261, or a nucleotide sequence that is at least or at least about 90%, at least 91% or at least about 91%, at least 92% or at least about 92%, at least 93% or at least about 93%, at least 94% or at least about 94%, at least 95% or at least about 95%, at least 96% or at least about 96%, at least 97% or at least about 97%, at least 98% or at least about 98%, or at least 99% or at least about 99% identical to SEQ ID NO:261. In some embodiments, the scFv comprises the amino acid sequence set forth in SEQ ID NO:6, or an amino acid sequence that is at least or at least about 90%, at least or at least about 91%, at least 92% or at least about 92%, at least 93% or at least about 93%, at least 94% or at least about 94%, at least 95% or at least about 95%, at least 96% or at least about 96%, at least 97% or at least about 97%, at least 98% or at least about 98%, or at least 99% or at least about 99% identical to SEQ ID NO:6.In some embodiments, the scFv is encoded by the nucleotide sequence set forth in SEQ ID NO:262, or a nucleotide sequence that is at least or at least about 90%, at least or at least about 91%, at least 92% or at least about 92%, at least 93% or at least about 93%, at least 94% or at least about 94%, at least 95% or at least about 95%, at least 96% or at least about 96%, at least 97% or at least about 97%, at least 98% or at least about 98%, or at least 99% or at least about 99% identical to SEQ ID NO:262.

[0260] Among the anti-GPRC5D CARs provided, the antibody or antigen-binding fragment comprises a V that includes an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 97%, at least 98%, or at least 98%, or at least 99% identity to SEQ ID NO:27. H and comprising the sequence set forth in SEQ ID NO:28, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:28. LIn some embodiments, the provided CARs include CARs in which the antibody or antigen-binding fragment comprises a V region, wherein the antibody or antigen-binding fragment comprises an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:27. H and comprising the sequence set forth in SEQ ID NO:66, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:66. L In some embodiments, the antibody or antigen-binding fragment of the CAR provided has a V region having CDRH1, CDRH2, and CDRH3 comprising the amino acid sequences of SEQ ID NOs: 120, 121, and 122, respectively. H and a V region having CDRL1, CDRL2, and CDRL3 comprising the amino acid sequences of SEQ ID NOs: 130, 131, and 132, respectively. L In some embodiments, the antibody or antigen-binding fragment of the provided CAR comprises a V region having a CDRH1, a CDRH2, and a CDRH3 comprising the amino acid sequences of SEQ ID NOs: 123, 124, and 122, respectively. Hand V comprising CDRL1, CDRL2, and CDRL3 comprising the amino acid sequences of SEQ ID NOs: 130, 131, and 132, respectively. L In some embodiments, the antibody or antigen-binding fragment of the provided CAR comprises a V region having a CDRH1, a CDRH2, and a CDRH3 comprising the amino acid sequences of SEQ ID NOs: 125, 126, and 122, respectively. H and V comprising CDRL1, CDRL2, and CDRL3 comprising the amino acid sequences of SEQ ID NOs: 130, 131, and 132, respectively. L In some embodiments, the antibody or antigen-binding fragment of the provided CAR comprises a V region having a CDRH1, a CDRH2, and a CDRH3 comprising the amino acid sequences of SEQ ID NOs: 127, 128, and 129, respectively. H and V comprising CDRL1, CDRL2, and CDRL3 comprising the amino acid sequences of SEQ ID NOs: 133, 134, and 132, respectively. L In some embodiments, the V H The region comprises the sequence shown in SEQ ID NO:27, L The region comprises the sequence set forth in SEQ ID NO: 28. H The region comprises the sequence shown in SEQ ID NO:27, LThe region comprises the sequence set forth in SEQ ID NO:66. In some embodiments, the antibody or antigen-binding fragment is a single-chain antibody fragment, e.g., an scFv. In some embodiments, the scFv comprises the amino acid sequence set forth in SEQ ID NO:7, or an amino acid sequence at least or at least about 90%, at least 91%, or at least about 91%, at least 92%, or at least about 92%, at least 93%, or at least about 93%, at least 94%, or at least about 94%, at least 95%, or at least about 95%, at least 96%, or at least about 96%, at least 97%, or at least about 97%, at least 98%, or at least about 98%, or at least 99% or at least about 99% identical to SEQ ID NO:7. In some embodiments, the scFv is encoded by the nucleotide sequence set forth in SEQ ID NO:263, or a nucleotide sequence that is at least or at least about 90%, at least or at least about 91%, at least 92% or at least about 92%, at least 93% or at least about 93%, at least 94% or at least about 94%, at least 95% or at least about 95%, at least 96% or at least about 96%, at least 97% or at least about 97%, at least 98% or at least about 98%, or at least 99% or at least about 99% identical to SEQ ID NO:263. In some embodiments, the scFv comprises the amino acid sequence set forth in SEQ ID NO:8, or an amino acid sequence that is at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identical to SEQ ID NO:8.In some embodiments, the scFv is encoded by the nucleotide sequence set forth in SEQ ID NO:264, or a nucleotide sequence that is at least or at least about 90%, at least or at least about 91%, at least 92% or at least about 92%, at least 93% or at least about 93%, at least 94% or at least about 94%, at least 95% or at least about 95%, at least 96% or at least about 96%, at least 97% or at least about 97%, at least 98% or at least about 98%, or at least 99% or at least about 99% identical to SEQ ID NO:264.

[0261] Among the anti-GPRC5D CARs provided, the antibody or antigen-binding fragment comprises a V that includes an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:29. H and comprising the sequence set forth in SEQ ID NO:30, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:30. LIn some embodiments, the provided CARs include CARs in which the antibody or antigen-binding fragment comprises a V region, wherein the antibody or antigen-binding fragment comprises an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:29. H and comprising a sequence set forth in SEQ ID NO:67, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:67. L In some embodiments, the provided CAR antibody or antigen-binding fragment has a V region having CDRH1, CDRH2, and CDRH3 comprising the amino acid sequences of SEQ ID NOs: 135, 136, and 137, respectively. H and a V region having CDRL1, CDRL2, and CDRL3 comprising the amino acid sequences of SEQ ID NOs: 145, 146, and 147, respectively. L In some embodiments, the antibody or antigen-binding fragment of the provided CAR comprises a V region having a CDRH1, a CDRH2, and a CDRH3 comprising the amino acid sequences of SEQ ID NOs: 138, 139, and 137, respectively. Hand V comprising CDRL1, CDRL2, and CDRL3 comprising the amino acid sequences of SEQ ID NOs: 145, 146, and 147, respectively. L In some embodiments, the antibody or antigen-binding fragment of the provided CAR comprises a V region having a CDRH1, a CDRH2, and a CDRH3 comprising the amino acid sequences of SEQ ID NOs: 140, 141, and 137, respectively. H and V comprising CDRL1, CDRL2, and CDRL3 comprising the amino acid sequences of SEQ ID NOs: 145, 146, and 147, respectively. L In some embodiments, the antibody or antigen-binding fragment of the provided CAR comprises a V region having a CDRH1, a CDRH2, and a CDRH3 comprising the amino acid sequences of SEQ ID NOs: 142, 143, and 144, respectively. H and V comprising CDRL1, CDRL2, and CDRL3 comprising the amino acid sequences of SEQ ID NOs: 148, 149, and 147, respectively. L In some embodiments, the V H The region comprises the sequence shown in SEQ ID NO:29, L The region comprises the sequence set forth in SEQ ID NO: 30. H The region comprises the sequence shown in SEQ ID NO:29, LThe region comprises the sequence set forth in SEQ ID NO:67. In some embodiments, the antibody or antigen-binding fragment is a single-chain antibody fragment, e.g., an scFv. In some embodiments, the scFv comprises the amino acid sequence set forth in SEQ ID NO:9, or an amino acid sequence at least or at least about 90%, at least 91%, or at least about 91%, at least 92%, or at least about 92%, at least 93%, or at least about 93%, at least 94%, or at least about 94%, at least 95%, or at least about 95%, at least 96%, or at least about 96%, at least 97%, or at least about 97%, at least 98%, or at least about 98%, or at least 99% or at least about 99% identical to SEQ ID NO:9. In some embodiments, the scFv is encoded by the nucleotide sequence set forth in SEQ ID NO:265, or a nucleotide sequence that is at least or at least about 90%, at least or at least about 91%, at least 92% or at least about 92%, at least 93% or at least about 93%, at least 94% or at least about 94%, at least 95% or at least about 95%, at least 96% or at least about 96%, at least 97% or at least about 97%, at least 98% or at least about 98%, or at least 99% or at least about 99% identical to SEQ ID NO:265. In some embodiments, the scFv comprises the amino acid sequence set forth in SEQ ID NO:10, or an amino acid sequence that is at least or at least about 90%, at least or at least about 91%, at least 92% or at least about 92%, at least 93% or at least about 93%, at least 94% or at least about 94%, at least 95% or at least about 95%, at least 96% or at least about 96%, at least 97% or at least about 97%, at least 98% or at least about 98%, or at least 99% or at least about 99% identical to SEQ ID NO:10.In some embodiments, the scFv is encoded by the nucleotide sequence set forth in SEQ ID NO:266, or a nucleotide sequence that is at least or at least about 90%, at least or at least about 91%, at least 92% or at least about 92%, at least 93% or at least about 93%, at least 94% or at least about 94%, at least 95% or at least about 95%, at least 96% or at least about 96%, at least 97% or at least about 97%, at least 98% or at least about 98%, or at least 99% or at least about 99% identical to SEQ ID NO:266.

[0262] Among the anti-GPRC5D CARs provided, the antibody or antigen-binding fragment comprises a V that includes an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to the sequence set forth in SEQ ID NO:31, or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:31. H and comprising the sequence set forth in SEQ ID NO:32, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:32. LIn some embodiments, the provided CARs include CARs in which the antibody or antigen-binding fragment comprises a V region, wherein the antibody or antigen-binding fragment comprises an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to the sequence set forth in SEQ ID NO:31, or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:31. H and comprising the sequence set forth in SEQ ID NO:68, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:68. L In some embodiments, the antibody or antigen-binding fragment of the CAR provided has a V region having CDRH1, CDRH2, and CDRH3 comprising the amino acid sequences of SEQ ID NOs: 135, 150, and 151, respecti...

Claims

1. (1) An extracellular antigen-binding domain that specifically binds to human G protein-coupled receptor class C group 5 member D (GPRC5D), (i) a heavy chain variable (V) comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:21, 23, 25, 27, 29, 31, or 33; H ) area; and (ii) a light chain variable (V) comprising an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:22, 24, 26, 28, 30, 32, 34, 63, 64, 65, 66, 67, 68, or 69; L )region an extracellular antigen-binding domain comprising: (2) a spacer at least 125 amino acids in length; (3) a transmembrane domain; and (4) Intracellular signal transduction region A chimeric antigen receptor comprising:

2. (1) An extracellular antigen-binding domain that specifically binds to human G protein-coupled receptor class C group 5 member D (GPRC5D), (i) V selected from SEQ ID NOs: 21, 23, 25, 27, 29, 31, and 33 H heavy chain variable (V) domains, including heavy chain complementarity determining region 1 (CDR-H1), heavy chain complementarity determining region 2 (CDR-H2), and heavy chain complementarity determining region 3 (CDR-H3), which are contained within the amino acid sequence of the V H ) area; and (ii) V selected from SEQ ID NOs: 22, 24, 26, 28, 30, 32, 34, 63, 64, 65, 66, 67, 68, and 69 L The light chain variable (V) region contains the light chain complementarity determining region 1 (CDR-L1), the light chain complementarity determining region 2 (CDR-L2), and the light chain complementarity determining region 3 (CDR-L3), which are contained within the amino acid sequence of the V L )region an extracellular antigen-binding domain comprising: (2) a spacer at least 125 amino acids in length; (3) a transmembrane domain; and (4) Intracellular signal transduction region A chimeric antigen receptor comprising:

3. (1) An extracellular antigen-binding domain that specifically binds to human G protein-coupled receptor class C group 5 member D (GPRC5D), (i) a CDR-H1 comprising an amino acid sequence selected from SEQ ID NOs: 75, 78, 80, 90, 93, 95, 105, 108, 110, 120, 123, 125, 135, 138, 140, 152, 162, 165, and 167; (b) a CDR-H2 comprising an amino acid sequence selected from SEQ ID NOs: 76, 79, 81, 91, 94, 96, 106, 109, 111, 121, 124, 126, 136, 139, 141, 150, 153, 154, 163, 166, and 168; and (c) a CDR-H3 comprising an amino acid sequence selected from SEQ ID NOs: 77, 92, 107, 122, 137, 151, and 164. H ) area; and (ii) a CDR-L1 comprising an amino acid sequence selected from SEQ ID NOs: 85, 100, 115, 130, 145, 157, and 172; (b) a CDR-L2 comprising an amino acid sequence selected from SEQ ID NOs: 86, 101, 116, 131, 146, and 158; and (c) a CDR-L3 comprising an amino acid sequence selected from SEQ ID NOs: 87, 102, 117, 132, 147, 159, 173, and 297. L )region an extracellular antigen-binding domain comprising: (2) a spacer at least 125 amino acids in length; (3) a transmembrane domain; and (4) Intracellular signal transduction region A chimeric antigen receptor comprising:

4. the extracellular antigen-binding domain (i) a heavy chain variable (V) comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:21, 23, 25, 27, 29, 31, or 33; H ) area; and (ii) a light chain variable (V) comprising an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:22, 24, 26, 28, 30, 32, 34, 63, 64, 65, 66, 67, 68, or 69; L )region 4. The chimeric antigen receptor of claim 2 or claim 3, comprising:

5. The spacer may be 125 to 300 or about 125 to about 300, 125 to 250 or about 125 to about 250, 125 to 230 or about 125 to about 230, 125 to 200 or about 125 to about 200, 125 to 180 or about 125 to about 180, 125 to 150 or about 125 to about 150, 150 to 300 or about 150 to about 300, 150 to 250 or about 150 to about 250, 150 to 230 or about 150 to about 230, 150 to 200 or about 150 to about 200, 150 to 180 or about 150 to about 180, 180 to 300, or The chimeric antigen receptor of any one of claims 1 to 4, having a length of about 180 to about 300, 180 to 250, or about 180 to about 250, 125 to 300, or about 125 to about 300, 180 to 230, or about 180 to about 230, 180 to 200, or about 180 to about 200, 200 to 300, or about 200 to about 250, 200 to 230, or about 200 to about 230, 230 to 300, or about 230 to about 250, or 250 to 300.

6. The spacer is 130 amino acids or at least about 130 amino acids in length, 140 amino acids or at least about 140 amino acids in length, 150 amino acids or at least about 150 amino acids in length, 160 amino acids or at least about 160 amino acids in length, 170 amino acids or at least about 170 amino acids in length, 180 amino acids or at least about 180 amino acids in length, 190 amino acids or at least about 190 amino acids in length, 200 amino acids or at least about 200 amino acids in length, 210 amino acids or at least about 210 amino acids in length, or 220 amino acids or at least about 220 amino acids in length. is 221 amino acids in length, or at least about 221 amino acids in length, 222 amino acids in length, or at least about 222 amino acids in length, 223 amino acids in length, or at least about 223 amino acids in length, 224 amino acids in length, 225 amino acids in length, or at least about 225 amino acids in length, 226 amino acids in length, 227 amino acids in length, 228 amino acids in length, or 229 amino acids in length, or having a length between any of the foregoing; or The spacer is about or at least about 130 amino acids in length, about or at least about 140 amino acids in length, about or at least about 150 amino acids in length, about or at least about 160 amino acids in length, about or at least about 170 amino acids in length, about or at least about 180 amino acids in length, about or at least about 190 amino acids in length, about or at least about 200 amino acids in length, about or at least about 210 amino acids in length, or about or at least about 220 amino acids in length. or at least about 221 amino acids in length, about 222 amino acids in length, about 223 amino acids in length, about 224 amino acids in length, about 225 amino acids in length, about 226 amino acids in length, about 227 amino acids in length, about 228 amino acids in length, or about 229 amino acids in length, or having a length between any of the foregoing. The chimeric antigen receptor of any one of claims 1 to 5.

7. The chimeric antigen receptor of any one of claims 1 to 6, wherein the spacer comprises a portion of an immunoglobulin.

8. The chimeric antigen receptor of any one of claims 1 to 7, wherein the spacer comprises sequences of a hinge region, a CH2 region, and a CH3 region.

9. the hinge region comprises all or a portion of an IgG4 hinge region and / or an IgG2 hinge region, wherein the IgG4 hinge region is optionally a human IgG4 hinge region and the IgG2 hinge region is optionally a human IgG2 hinge region; Said C H 2 domains IgG4 C H 2 and / or IgG2 C H 2, wherein the IgG4 C H 2 is optionally human IgG4 C H 2, and the IgG2 C H 2 is optionally human IgG2 C H 2; and / or Said C H 3 regions IgG4 C H 3 and / or IgG2 C H 3, wherein the IgG4 C H 3 is optionally human IgG4 C H 3, and the IgG2 C H 3 is optionally human IgG2 C H 3, The chimeric antigen receptor of claim 8.

10. The hinge region, CH2, and CH3 are all or a part of a hinge derived from human IgG4, H All or part of 2, and C H 10. The chimeric antigen receptor of claim 8 or claim 9, comprising all or a portion of 3.

11. Said hinge area, C H 2, and C H 3 are chimeric, including hinges derived from human IgG4 and human IgG2, C H 2, and C H 10. The chimeric antigen receptor of claim 8 or claim 9, comprising:

12. The spacer is an IgG4 / 2 chimeric hinge region, or a modified IgG4 hinge region comprising at least one amino acid substitution compared to a human IgG4 hinge; an IgG2 / 4 chimeric hinge region; H 2 domains; and IgG4 C H The chimeric antigen receptor of any one of claims 1 to 11, comprising three regions.

13. The spacer is (i) the sequence shown in SEQ ID NO: 17; (ii) a functional variant of SEQ ID NO:17 having at least 95%, at least 96%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% or at least 99% sequence identity to SEQ ID NO:17; or (iii) a continuous portion of (i) or (ii) that is at least 125 amino acids in length 13. The chimeric antigen receptor of any one of claims 1 to 12, which is or comprises:

14. The chimeric antigen receptor of any one of claims 1 to 13, wherein the spacer is or comprises the sequence shown in SEQ ID NO:

17.

15. 15. The chimeric antigen receptor of any one of claims 1 to 14, wherein the spacer is or comprises an amino acid sequence encoded by the nucleotide sequence shown in SEQ ID NO:

74.

16. The above V H wherein the region comprises an amino acid sequence set forth in SEQ ID NO:21, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:21; and L the region comprises the amino acid sequence set forth in SEQ ID NO:22, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:22; The above V H wherein the region comprises an amino acid sequence set forth in SEQ ID NO:21, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:21; and L the region comprises the amino acid sequence set forth in SEQ ID NO:63, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:63; The above V H wherein the V region comprises an amino acid sequence set forth in SEQ ID NO:23, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:23; and L the region comprises the amino acid sequence set forth in SEQ ID NO:24, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:24; The above V H wherein the V region comprises an amino acid sequence set forth in SEQ ID NO:23, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:23; and L the region comprises the amino acid sequence set forth in SEQ ID NO:64, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:64; The above V H wherein the region comprises an amino acid sequence set forth in SEQ ID NO:25, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:25; and L the region comprises the amino acid sequence set forth in SEQ ID NO:26, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:26; The above V H wherein the region comprises an amino acid sequence set forth in SEQ ID NO:25, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:25; and L the region comprises the amino acid sequence set forth in SEQ ID NO:65, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:65; The above V H wherein the region comprises an amino acid sequence set forth in SEQ ID NO:27, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:27; and L the region comprises the amino acid sequence set forth in SEQ ID NO:28, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:28; The above V H wherein the region comprises an amino acid sequence set forth in SEQ ID NO:27, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:27; and L the region comprises the amino acid sequence set forth in SEQ ID NO:66, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:66; The above V H wherein the V region comprises an amino acid sequence set forth in SEQ ID NO:29, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:29; and L the region comprises the amino acid sequence set forth in SEQ ID NO:30, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:30; The above V H wherein the V region comprises an amino acid sequence set forth in SEQ ID NO:29, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:29; and L the region comprises the amino acid sequence set forth in SEQ ID NO:67, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:67; The above V H wherein the region comprises an amino acid sequence set forth in SEQ ID NO:31, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:31; and L the region comprises the amino acid sequence set forth in SEQ ID NO:32, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:32; The above V H wherein the region comprises an amino acid sequence set forth in SEQ ID NO:31, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:31; and L the region comprises the amino acid sequence set forth in SEQ ID NO:68, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:68; The above V H wherein the region comprises an amino acid sequence set forth in SEQ ID NO:33, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:33; and L the region comprises the amino acid sequence set forth in SEQ ID NO:34, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:34; or The above V H wherein the region comprises an amino acid sequence set forth in SEQ ID NO:33, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:33; and L the region comprises an amino acid sequence set forth in SEQ ID NO:69, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:69; The chimeric antigen receptor of any one of claims 1 to 15.

17. The above V H The V region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 80, 81, and 77, respectively; L whether the regions comprise the amino acid sequences of SEQ ID NOs: 85, 86, and 87, respectively; The above V H The V region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 95, 96, and 92, respectively; L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 100, 101, and 102, respectively; The above V H The V region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 110, 111, and 107, respectively; L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 115, 116, and 117, respectively; The above V H The V region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 125, 126, and 122, respectively; L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 130, 131, and 132, respectively; The above V H The V region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 140, 141, and 137, respectively; L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 145, 146, and 147, respectively; The above V H wherein the V region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 140, 154, and 151, respectively; L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 157, 158, and 159, respectively; The above V H wherein the V region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 167, 168, and 164, respectively; L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 172, 86, and 173, respectively; or The above V H wherein the V region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 169, 170, and 171, respectively; L The region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 174, 89, and 297, respectively; The chimeric antigen receptor of any one of claims 1 to 16.

18. The above V H Region and V L the region comprises the amino acid sequence set forth in SEQ ID NOs:21 and 22, respectively, or the amino acid sequence set forth in SEQ ID NOs:21 and 63, respectively; The above V H Region and V L the region comprises the amino acid sequence set forth in SEQ ID NOs:23 and 24, respectively, or the amino acid sequence set forth in SEQ ID NOs:23 and 64, respectively; The above V H Region and V L the region comprises the amino acid sequence set forth in SEQ ID NOs:25 and 26, respectively, or the amino acid sequence set forth in SEQ ID NOs:25 and 65, respectively; The above V H Region and V L the region comprises the amino acid sequence set forth in SEQ ID NOs:27 and 28, respectively, or the amino acid sequence set forth in SEQ ID NOs:27 and 66, respectively; The above V H Region and V L whether the region comprises the amino acid sequence set forth in SEQ ID NOs:29 and 30, respectively, or the amino acid sequence set forth in SEQ ID NOs:29 and 67, respectively; The above V H Region and V L the region comprises the amino acid sequence set forth in SEQ ID NOs:31 and 32, respectively, or the amino acid sequence set forth in SEQ ID NOs:31 and 68, respectively; or The above V H Region and V L The region comprises the amino acid sequence set forth in SEQ ID NOs: 33 and 34 or the amino acid sequence set forth in SEQ ID NOs: 33 and 69, respectively; The chimeric antigen receptor of any one of claims 1 to 17.

19. The above V H wherein the region comprises an amino acid sequence set forth in SEQ ID NO:21, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:21; and L the region comprises the amino acid sequence set forth in SEQ ID NO:22, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:22; The above V H wherein the region comprises an amino acid sequence set forth in SEQ ID NO:21, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:21; and L the region comprises the amino acid sequence set forth in SEQ ID NO:63, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:63; The above V H wherein the V region comprises an amino acid sequence set forth in SEQ ID NO:23, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:23; and L the region comprises the amino acid sequence set forth in SEQ ID NO:24, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:24; The above V H wherein the V region comprises an amino acid sequence set forth in SEQ ID NO:23, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:23; and L the region comprises the amino acid sequence set forth in SEQ ID NO:64, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:64; The above V H wherein the region comprises an amino acid sequence set forth in SEQ ID NO:27, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:27; and L the region comprises the amino acid sequence set forth in SEQ ID NO:28, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:28; The above V H wherein the region comprises an amino acid sequence set forth in SEQ ID NO:27, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:27; and L the region comprises the amino acid sequence set forth in SEQ ID NO:66, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:66; The above V H wherein the region comprises an amino acid sequence set forth in SEQ ID NO:31, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:31; and L the region comprises the amino acid sequence set forth in SEQ ID NO:32, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least or at least about 91%, at least or at least about 92%, at least or at least about 92%, at least or at least about 93%, at least or at least about 94%, at least or at least about 94%, at least or at least about 95%, at least or at least about 96%, at least or at least about 96%, at least or at least about 97%, at least or at least about 97%, at least 98% or at least about 98%, or at least 99% or at least about 99% identity to SEQ ID NO:32; or The above V H wherein the region comprises an amino acid sequence set forth in SEQ ID NO:31, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:31; and L the region comprises an amino acid sequence set forth in SEQ ID NO:68, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:68; The chimeric antigen receptor of any one of claims 1 to 18.

20. The above V H wherein the V region comprises CDR-H1, CDR-H2, and CDR-H3 comprising the amino acid sequences of SEQ ID NOs: 80, 81, and 77, respectively; L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 85, 86, and 87, respectively; The above V H The V region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 95, 96, and 92, respectively; L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 100, 101, and 102, respectively; The above V H wherein the V region comprises CDR-H1, CDR-H2, and CDR-H3 comprising the amino acid sequences of SEQ ID NOs: 125, 126, and 122, respectively; L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 130, 131, and 132, respectively; or The above V H wherein the V region comprises CDR-H1, CDR-H2, and CDR-H3 comprising the amino acid sequences of SEQ ID NOs: 140, 154, and 151, respectively; L The regions include CDR-L1, CDR-L2, and CDR-L3, each of which comprises the amino acid sequences of SEQ ID NOs: 157, 158, and 159, respectively; The chimeric antigen receptor of any one of claims 1 to 19.

21. The above V H Region and V L the region comprises the amino acid sequence set forth in SEQ ID NOs:21 and 22, respectively, or the amino acid sequence set forth in SEQ ID NOs:21 and 63, respectively; The above V H Region and V L the region comprises the amino acid sequence set forth in SEQ ID NOs:23 and 24, respectively, or the amino acid sequence set forth in SEQ ID NOs:23 and 64, respectively; The above V H Region and V L the region comprises the amino acid sequence set forth in SEQ ID NOs:27 and 28, respectively, or the amino acid sequence set forth in SEQ ID NOs:27 and 66, respectively; or The above V H Region and V L The region comprises the amino acid sequence set forth in SEQ ID NOs: 31 and 32, respectively, or the amino acid sequence set forth in SEQ ID NOs: 31 and 68, respectively; The chimeric antigen receptor of any one of claims 1 to 20.

22. The above V H wherein the region comprises an amino acid sequence set forth in SEQ ID NO:27, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:27; and L the region comprises the amino acid sequence set forth in SEQ ID NO:28, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:28; or The above V H wherein the region comprises an amino acid sequence set forth in SEQ ID NO:27, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:27; and L the region comprises an amino acid sequence set forth in SEQ ID NO:66, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% identity to SEQ ID NO:66; The chimeric antigen receptor of any one of claims 1 to 21.

23. V as shown in SEQ ID NO:27 H heavy chain variable (V) domains, including CDR-H1, CDR-H2, and CDR-H3, contained within the amino acid sequence of the V H ) region; and the V region shown in SEQ ID NO: 28 or 66 L The light chain variable (V) region contains CDR-L1, CDR-L2, and CDR-L3. L 23. The chimeric antigen receptor of any one of claims 1 to 22, comprising a nucleotide sequence corresponding to a nucleotide sequence of ...

24. The above V H The V region comprises the amino acid sequence of SEQ ID NO: 125, 126 and 122, respectively, and L The chimeric antigen receptor of any one of claims 1 to 23, wherein the regions comprise the amino acid sequences of SEQ ID NOs: 130, 131 and 132, respectively.

25. The above V H The V region comprises the amino acid sequence of SEQ ID NO: 127, 128 and 129, respectively, and L The chimeric antigen receptor of any one of claims 1 to 23, wherein the regions comprise the amino acid sequences of SEQ ID NOs: 133, 134 and 132, respectively.

26. The above V H The V region comprises the amino acid sequence of SEQ ID NO: 120, 121 and 122, respectively, and L The chimeric antigen receptor of any one of claims 1 to 23, wherein the regions comprise the amino acid sequences of SEQ ID NOs: 130, 131 and 132, respectively.

27. The above V H The V region comprises the amino acid sequence of SEQ ID NO: 123, 124 and 122, respectively, and L The chimeric antigen receptor of any one of claims 1 to 23, wherein the regions comprise the amino acid sequences of SEQ ID NOs: 130, 131 and 132, respectively.

28. The above V H Region and V L 24. The chimeric antigen receptor of any one of claims 1 to 23, wherein the region comprises the amino acid sequence set forth in SEQ ID NOs: 27 and 28, respectively, or the amino acid sequence set forth in SEQ ID NOs: 27 and 66, respectively.

29. The chimeric antigen receptor of any one of claims 1 to 28, wherein the extracellular antigen-binding domain is a single-chain antibody fragment.

30. 30. The chimeric antigen receptor of any one of claims 1 to 29, wherein the single-chain antibody fragment is or comprises a single-chain variable fragment (scFv).

31. The above V H Region and V L The chimeric antigen receptor of any one of claims 1 to 30, wherein the regions are linked by a flexible linker.

32. The linker comprises the amino acid sequence 32. The chimeric antigen receptor of claim 31 , comprising:

33. The above V H The region V L The chimeric antigen receptor of any one of claims 1 to 32, wherein the chimeric antigen receptor is amino terminal to the region.

34. the extracellular antigen-binding domain comprises an amino acid sequence selected from SEQ ID NOs: 1, 3, 5, 7, 9, 11 and 13, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to an amino acid sequence selected from SEQ ID NOs: 1, 3, 5, 7, 9, 11 and 13; and / or the extracellular antigen-binding domain is encoded by a nucleotide sequence selected from SEQ ID NOs: 257, 259, 261, 263, 265, 267, and 269, or a nucleotide sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% or at least about 99% sequence identity to a nucleotide sequence selected from SEQ ID NOs: 257, 259, 261, 263, 265, 267, and 269; The chimeric antigen receptor of any one of claims 1 to 33.

35. 35. The chimeric antigen receptor of any one of claims 1 to 34, wherein the extracellular antigen-binding domain comprises an amino acid sequence selected from SEQ ID NOs: 1, 3, 5, 7, 9, 11 and 13.

36. 35. The chimeric antigen receptor of any one of claims 1 to 34, wherein the extracellular antigen-binding domain comprises an amino acid sequence selected from SEQ ID NOs: 1, 3, 7 and 11, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NOs: 1, 3, 7 and 11.

37. 37. The chimeric antigen receptor of any one of claims 1 to 36, wherein the antigen-binding domain comprises an amino acid sequence selected from SEQ ID NOs: 1, 3, 7 and 11.

38. 38. The chimeric antigen receptor of any one of claims 1 to 34 or 37, wherein the antigen-binding domain comprises the amino acid sequence set forth in SEQ ID NO:7, or an amino acid sequence having at least or at least about 90%, at least 91%, or at least about 91%, at least 92%, or at least about 92%, at least 93%, or at least about 93%, at least 94%, or at least about 94%, at least 95%, or at least about 95%, at least 96%, or at least about 96%, at least 97%, or at least about 97%, at least 98%, or at least about 98%, or at least 99% or at least about 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:

7.

39. The chimeric antigen receptor of any one of claims 1 to 38, wherein the antigen-binding domain comprises the amino acid sequence shown in SEQ ID NO:

7.

40. The above V H The region V L The chimeric antigen receptor of any one of claims 1 to 32, wherein the chimeric antigen receptor is carboxy-terminal to the region.

41. the extracellular antigen-binding domain comprises an amino acid sequence selected from SEQ ID NOs: 2, 4, 6, 8, 10, 12 and 14, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to an amino acid sequence selected from SEQ ID NOs: 2, 4, 6, 8, 10, 12 and 14; and / or the extracellular antigen-binding domain is encoded by a nucleotide sequence selected from SEQ ID NOs: 258, 260, 262, 264, 266, 268, and 270, or a nucleotide sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% or at least about 99% sequence identity to a nucleotide sequence selected from SEQ ID NOs: 258, 260, 262, 264, 266, 268, and 270; The chimeric antigen receptor of any one of claims 1 to 32 and 40.

42. 42. The chimeric antigen receptor of any one of claims 1 to 32, 40 and 41, wherein the extracellular antigen-binding domain comprises an amino acid sequence selected from SEQ ID NOs: 2, 4, 6, 8, 10, 12 and 14.

43. 42. The chimeric antigen receptor of any one of claims 1 to 32, 40 and 41, wherein the extracellular antigen-binding domain comprises an amino acid sequence selected from SEQ ID NOs: 2, 4, 8 and 12, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NOs: 2, 4, 8 and 12.

44. 44. The chimeric antigen receptor of any one of claims 1 to 32 or 40 to 43, wherein the extracellular antigen-binding domain comprises an amino acid sequence selected from SEQ ID NOs: 2, 4, 8 and 12.

45. 44. The chimeric antigen receptor of any one of claims 1 to 32, 40, 41 and 43, wherein the antigen-binding domain comprises the amino acid sequence set forth in SEQ ID NO:8 or an amino acid sequence having at least or at least about 90%, at least 91%, or at least about 91%, at least 92%, or at least about 92%, at least 93%, or at least about 93%, at least 94%, or at least about 94%, at least 95%, or at least about 95%, at least 96%, or at least about 96%, at least 97%, or at least about 97%, at least 98%, or at least about 98%, or at least 99% or at least about 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:

8.

46. 46. ​​The chimeric antigen receptor of any one of claims 1 to 32 or 40 to 45, wherein the extracellular antigen-binding domain comprises the amino acid sequence shown in SEQ ID NO:

8.

47. The chimeric antigen receptor of any one of claims 1 to 46, wherein the intracellular signaling region comprises an intracellular cytoplasmic signaling domain.

48. 48. The chimeric antigen receptor of claim 47, wherein the intracellular signaling domain is or comprises the cytoplasmic signaling domain of the CD3-zeta (CD3ζ) chain or a functional variant or signaling portion thereof.

49. 49. The chimeric antigen receptor of any one of claims 47 or 48, wherein the intracellular signaling domain is or comprises an amino acid sequence set forth in SEQ ID NO:20, or an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% or at least about 99% sequence identity to SEQ ID NO:

20.

50. The chimeric antigen receptor of any one of claims 47 to 49, wherein the intracellular signaling region further comprises a costimulatory signaling region.

51. 51. The chimeric antigen receptor of claim 50, wherein the costimulatory signaling region comprises the intracellular signaling domain of CD28, 4-1BB, or ICOS, or a signaling portion thereof.

52. 52. The chimeric antigen receptor of claim 50 or 51, wherein the costimulatory signaling region comprises the intracellular signaling domain of CD28.

53. 53. The chimeric antigen receptor of any one of claims 50-52, wherein the costimulatory signaling region is or comprises an amino acid sequence set forth in SEQ ID NO:46 or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:

46.

54. 52. The chimeric antigen receptor of claim 50 or 51, wherein the costimulatory signaling region comprises the intracellular signaling domain of 4-1BB.

55. 55. The chimeric antigen receptor of any one of claims 50, 51 and 54, wherein the costimulatory signaling region is or comprises an amino acid sequence set forth in SEQ ID NO:19 or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:

19.

56. 56. The chimeric antigen receptor of any one of claims 50 to 55, wherein the costimulatory signaling region is between the transmembrane domain and the intracellular signaling region.

57. 57. The chimeric antigen receptor of any one of claims 1 to 56, wherein the transmembrane domain is or comprises a transmembrane domain from CD4, CD28, or CD8.

58. 58. The chimeric antigen receptor of any one of claims 1 to 57, wherein said transmembrane domain is or comprises a transmembrane domain derived from CD28.

59. 59. The chimeric antigen receptor of any one of claims 1 to 58, wherein the transmembrane domain is or comprises the amino acid sequence set forth in SEQ ID NO:18 or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:

18.

60. (1) An extracellular antigen-binding domain that specifically binds to human G protein-coupled receptor class C group 5 member D (GPRC5D), (i) a heavy chain variable (V) antibody comprising an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:27; H ) area; and (ii) a light chain variable (V) comprising an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:28 or 66; L )region an extracellular antigen-binding domain comprising: (2) IgG4 / 2 chimeric hinge or modified IgG4 hinge; IgG2 / 4 chimeric C H 2 domains; and IgG4 C H a spacer comprising three regions and optionally about 228 amino acids in length or the spacer set forth in SEQ ID NO: 17; (3) a transmembrane domain derived from human CD28; and (4) an intracellular signaling region containing the cytoplasmic signaling domain of the CD3-zeta (CD3ζ) chain and the intracellular signaling domain of the T cell costimulatory molecule; A chimeric antigen receptor comprising:

61. The above V H The region is V shown in SEQ ID NO:27 H CDR-H1, CDR-H2, and CDR-H3 contained within the V region amino acid sequence, L The region is V shown in SEQ ID NO: 28 or 66 L Contains CDR-L1, CDR-L2, and CDR-L3 contained within the region amino acid sequence; The above V H the regions comprising CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 125, 126, and 122, respectively; and L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 130, 131, and 132, respectively; The above V H wherein the V region comprises CDR-H1, CDR-H2, and CDR-H3 comprising the amino acid sequences of SEQ ID NOs: 120, 121, and 122, respectively; L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 130, 131, and 132, respectively; or The above V H wherein the V region comprises CDR-H1, CDR-H2, and CDR-H3 comprising the amino acid sequences of SEQ ID NOs: 123, 124, and 122, respectively; L The regions include CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences of SEQ ID NOs: 130, 131, and 132, respectively; 61. The chimeric antigen receptor of claim 60.

62. (1) An extracellular antigen-binding domain that specifically binds to human G protein-coupled receptor class C group 5 member D (GPRC5D), V as shown in SEQ ID NO:27 H V including CDR-H1, CDR-H2, and CDR-H3 contained within the amino acid sequence of the region H region; and V as shown in SEQ ID NO: 28 or 66 L The light chain variable (V) region contains CDR-L1, CDR-L2, and CDR-L3. L ) area; or V comprising CDR-H1, CDR-H2, and CDR-H3 containing the amino acid sequences of SEQ ID NOs: 125, 126, and 122, respectively. H V region, and CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 130, 131, and 132, respectively. L region; V comprising CDR-H1, CDR-H2, and CDR-H3 containing the amino acid sequences of SEQ ID NOs: 120, 121, and 122, respectively. H V region, and CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 130, 131, and 132, respectively. L Area; or V comprising CDR-H1, CDR-H2, and CDR-H3 containing the amino acid sequences of SEQ ID NOs: 123, 124, and 122, respectively. H V region, and CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 130, 131, and 132, respectively. L region an extracellular antigen-binding domain comprising: (2) IgG4 / 2 chimeric hinge or modified IgG4 hinge; IgG2 / 4 chimeric C H 2 domains; and IgG4 C H a spacer comprising three regions and optionally about 228 amino acids in length or the spacer set forth in SEQ ID NO: 17; (3) a transmembrane domain derived from human CD28; and (4) an intracellular signaling region comprising the cytoplasmic signaling domain of the human CD3-zeta (CD3ζ) chain and the intracellular signaling domain of human CD28 or human 4-1BB; A chimeric antigen receptor comprising:

63. The extracellular antigen-binding domain is V as shown in SEQ ID NO:27 H The amino acid sequence of the region and V shown in SEQ ID NO: 28 or 66 L and / or the extracellular antigen-binding domain comprises an scFv as set forth in SEQ ID NO:7 or SEQ ID NO:8; 63. The chimeric antigen receptor of any one of claims 60 to 62.

64. 64. The chimeric antigen receptor of any one of claims 60 to 63, wherein the transmembrane domain is or comprises the amino acid sequence set forth in SEQ ID NO: 18 or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to SEQ ID NO:

18.

65. the intracellular signaling region is selected from the group consisting of: (a) the amino acid sequence set forth in SEQ ID NO:20, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to SEQ ID NO:20; and (b) the amino acid sequence set forth in SEQ ID NO:46, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:

46.

65. The chimeric antigen receptor of any one of claims 60 to 64, comprising an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least or at least about 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% or at least about 99% sequence identity to the sequence set forth in SEQ ID NO:

46.

66. 66. The chimeric antigen receptor of any one of claims 60 to 65, wherein the intracellular signaling region is or comprises the sequence shown in SEQ ID NO: 20 and the sequence shown in SEQ ID NO:

46.

67. the intracellular signaling region is selected from the group consisting of: (a) the amino acid sequence set forth in SEQ ID NO:20, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:20; and (b) the amino acid sequence set forth in SEQ ID NO:19, or an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:

20.

65. The chimeric antigen receptor of any one of claims 60 to 64, comprising an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least or at least about 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% or at least about 99% sequence identity to the sequence set forth in SEQ ID NO:

19.

68. 68. A polynucleotide comprising a nucleotide sequence encoding the chimeric antigen receptor of any one of claims 1 to 67.

69. 69. The polynucleotide of claim 68, wherein the nucleic acid encoding the spacer comprises at least one modified splice donor site and / or modified splice acceptor site, wherein the modified splice donor site and / or modified splice acceptor site comprises one or more nucleotide modifications corresponding to a reference splice donor site and / or reference splice acceptor site contained in the sequence set forth in SEQ ID NO:

73.

70. 70. The polynucleotide of claim 69, wherein said one or more nucleotide modifications comprise an amino acid substitution.

71. the reference splice donor and / or reference splice acceptor site has a splice site prediction score of at least or at least about 0.4, at least 0.5 or at least about 0.5, at least 0.6 or at least about 0.6, at least 0.70 or at least about 0.70, at least 0.75 or at least about 0.75, at least 0.80 or at least about 0.80, at least 0.85 or at least about 0.85, at least 0.90 or at least about 0.90, at least 0.95 or at least about 0.95, at least 0.99 or at least about 0.99, or at least 1.0 or at least about 1.0; and / or the reference splice donor and / or reference splice acceptor site is predicted to be involved in a splice event at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or at least 100% of the time, 71. The polynucleotide of claim 69 or 70.

72. the reference splice donor site has the sequence and / or the reference splice acceptor site has the sequence Including, 72. The polynucleotide of any one of claims 69 to 71.

73. the reference splice donor site has the sequence and / or the reference splice acceptor site has the sequence Including, 73. The polynucleotide of any one of claims 69 to 72.

74. 74. The polynucleotide of any one of claims 69 to 73, wherein said one or more nucleotide modifications are silent and / or result in degenerate codons compared to SEQ ID NO: 73 and / or do not change the amino acid sequence of the encoded spacer.

75. the modified splice donor site and / or the modified splice acceptor site As shown in 75. The polynucleotide of any one of claims 69 to 74.

76. the modified splice donor site and / or the modified acceptor site 76. The polynucleotide of any one of claims 69 to 75, as set forth in

77. 77. The polynucleotide of any one of claims 69 to 76, wherein said spacer is encoded by the nucleotide sequence set forth in SEQ ID NO:

74.

78. 78. The polynucleotide of any one of claims 68 to 77, wherein upon expression in a cell, RNA, optionally messenger RNA (mRNA), transcribed from said polynucleotide exhibits reduced heterogeneity compared to the heterogeneity of mRNA transcribed from a reference polynucleotide, said reference polynucleotide encoding the same amino acid sequence as said polynucleotide, said reference polynucleotide differing from said polynucleotide in the presence of one or more splice donor sites and / or one or more splice acceptor sites in a nucleic acid encoding a spacer, and / or comprising one or more nucleotide alterations compared to said polynucleotide, and / or comprising the spacer set forth in SEQ ID NO:

73.

79. 79. The polynucleotide of any one of claims 68-78, wherein the polynucleotide is codon-optimized for expression in a human cell.

80. 79. The polynucleotide of any one of claims 68-78, wherein said chimeric antigen receptor is a first chimeric antigen receptor, and said polynucleotide further comprises a nucleotide sequence encoding a second chimeric antigen receptor.

81. 81. The polynucleotide of claim 80, wherein said first chimeric antigen receptor and said second chimeric antigen receptor are separated by one or more multicistronic elements.

82. 82. The polynucleotide of claim 81, wherein said one or more multicistronic elements is or comprises a ribosomal skipping sequence, optionally wherein said ribosomal skipping sequence is a T2A, P2A, E2A, or F2A element.

83. 83. The polynucleotide of claim 82, wherein the nucleotide sequence encoding said one or more multicistronic elements is a codon divergent form.

84. 84. The polynucleotide of claim 82 or claim 83, wherein the nucleotide sequence encoding the one or more multicistronic elements is or comprises the sequence set forth in SEQ ID NO:

319.

85. 85. The polynucleotide of any one of claims 80-84, wherein said second chimeric antigen receptor (CAR) comprises an extracellular antigen-binding domain that specifically binds to a second antigen expressed on or associated with multiple myeloma.

86. 86. The polynucleotide of claim 85, wherein said second CAR further comprises a spacer, a transmembrane domain, and an intracellular signaling region.

87. 87. The polynucleotide of claim 85 or claim 86, wherein the second antigen is selected from B-cell maturation antigen (BCMA), CD38, CD138, CS-1, BAFF-R, TACI, and FcRH5.

88. 88. The polynucleotide of any one of claims 85 to 87, wherein said second antigen is BCMA.

89. The second CAR is (1) An extracellular antigen-binding domain that specifically binds to BCMA, (i) a heavy chain variable (V) comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:189, 191, 193, 195, or 197; H ) area; and (ii) a light chain variable (V) comprising an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:190, 192, 194, 196, or 198; L )region an extracellular antigen-binding domain comprising: (2) Spacer, optionally an IgG4 / 2 chimeric hinge or a modified IgG4 hinge; IgG2 / 4 chimeric C H 2 domains; and IgG4 C H a spacer comprising three regions and optionally about 228 amino acids in length or the spacer set forth in SEQ ID NO: 17; (3) a transmembrane domain; and (4) Intracellular signal transduction region 89. The polynucleotide of any one of claims 85 to 88, comprising:

90. V of the second car H The region is V as set forth in SEQ ID NO: 189, 191, 193, 195, or 197. H The second CAR comprises CDR-H1, CDR-H2, and CDR-H3 contained within the amino acid sequence of the second CAR; and L The region is selected from the group consisting of V, VL and VL sequences shown in SEQ ID NO: 190, 192, 194, 196, or 198. L 90. The polynucleotide of claim 89, comprising CDR-L1, CDR-L2, and CDR-L3 contained within the region amino acid sequence.

91. The second CAR is (1) An extracellular antigen-binding domain that specifically binds to BCMA, (i) a heavy chain complementary determining region 1 (CDR-H1) comprising an amino acid sequence selected from SEQ ID NOs: 199, 202, 206, and 209; (b) a heavy chain complementary determining region 2 (CDR-H2) comprising an amino acid sequence selected from SEQ ID NOs: 200, 203, 207, and 210; and (c) a heavy chain complementary determining region 3 (CDR-H3) comprising an amino acid sequence selected from SEQ ID NOs: 201, 204, 205, 208, and 211. H ) area; and (ii) a light chain variable (V) antibody comprising: (i) a light chain complementarity determining region 1 (CDR-L1) comprising an amino acid sequence selected from SEQ ID NOs: 218, 221, 224, 227, and 230; (b) a light chain complementarity determining region 2 (CDR-L2) comprising an amino acid sequence selected from any one of SEQ ID NOs: 219, 222, 225, 228, and 231; and (c) a light chain complementarity determining region 3 (CDR-L3) comprising an amino acid sequence selected from SEQ ID NOs: 220, 223, and 226. L )region an extracellular antigen-binding domain comprising: (2) Spacer, optionally an IgG4 / 2 chimeric hinge or a modified IgG4 hinge; IgG2 / 4 chimeric C H 2 domains; and IgG4 C H a spacer comprising 3 regions, optionally at or about 228 amino acids in length, or the spacer set forth in SEQ ID NO: 17; (3) a transmembrane domain; and (4) Intracellular signal transduction region 89. The polynucleotide of any one of claims 85 to 88, comprising:

92. V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 199, 200, and 201, respectively, and L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 218, 219, and 220, respectively; V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 202, 203, and 204, respectively, and L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 221, 222, and 223, respectively; V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 199, 200, and 205, respectively, and L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 224, 225, and 226, respectively; V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 206, 207, and 208, respectively, and L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 227, 228, and 229, respectively; or V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and L The regions include CDR-L1, CDR-L2, and CDR-L3, each of which comprises the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; 92. The polynucleotide of any one of claims 89 to 91.

93. V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and L The regions include CDR-L1, CDR-L2, and CDR-L3, each of which comprises the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; 92. The polynucleotide of any one of claims 89 to 91.

94. V of the second car H Area and V L a region having (a) the amino acid sequence set forth in SEQ ID NO:189 and SEQ ID NO:190, respectively; or (b) an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:189; and a region having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98% sequence identity to SEQ ID NO:

190. comprises an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to NO:190; V of the second car H Area and V L a region having (a) the amino acid sequence set forth in SEQ ID NO:191 and SEQ ID NO:192, respectively; or (b) an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:191; and a region having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:

192. or comprises an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to NO:192; V of the second car H Area and V L a region having (a) the amino acid sequence set forth in SEQ ID NO:193 and SEQ ID NO:194, respectively, or (b) an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:193, and comprises an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to NO:194; V of the second car H Area and V L a region having (a) the amino acid sequence set forth in SEQ ID NO:195 and SEQ ID NO:196, respectively, or (b) an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:195, and or comprises an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to NO:196; or V of the second car H Area and V L a region having (a) the amino acid sequence set forth in SEQ ID NO:197 and SEQ ID NO:198, respectively, or (b) an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:197, and comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to NO:198; 94. The polynucleotide of any one of claims 89 to 93.

95. V of the second car H Area and V L the regions comprise the amino acid sequences set forth in SEQ ID NO:189 and SEQ ID NO:190, respectively; V of the second car H Area and V L whether the region comprises the amino acid sequence set forth in SEQ ID NO:191 and SEQ ID NO:192; V of the second car H Area and V L whether the region comprises the amino acid sequence set forth in SEQ ID NO:193 and SEQ ID NO:194; V of the second car H Area and V L The region comprises the amino acid sequence set forth in SEQ ID NO:195 and SEQ ID NO:196; and V of the second car H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NO: 197 and SEQ ID NO: 198, respectively; 95. The polynucleotide of any one of claims 89 to 94.

96. V of the second car H The region V L 96. The polynucleotide of any one of claims 89-95, wherein said polynucleotide is amino terminal to said region.

97. V of the second car H The region V L 96. The polynucleotide of any one of claims 89-95, wherein said polynucleotide is carboxy terminal to said region.

98. 98. The polynucleotide of any one of claims 89-97, wherein the antigen-binding domain of the second CAR comprises an amino acid sequence selected from SEQ ID NOs: 237, 238, 239, 240, and 241, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NOs: 237, 238, 239, 240, and 241.

99. 99. The polynucleotide of any one of claims 89-98, wherein the antigen-binding domain of the second CAR comprises an amino acid sequence selected from SEQ ID NOs: 237, 238, 239, 240, and 241.

100. V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and L the regions comprise CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; and / or V of the second car H Area and V L the regions comprise the amino acid sequences set forth in SEQ ID NO:197 and SEQ ID NO:198, respectively; and / or the extracellular antigen-binding domain of the second CAR comprises the amino acid sequence set forth in SEQ ID NO:241, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:241; 99. The polynucleotide of any one of claims 89 to 99.

101. 101. The polynucleotide of any one of claims 86-100, wherein the transmembrane domain of said second CAR is or comprises a transmembrane domain derived from CD4, CD28, or CD8, optionally a transmembrane domain derived from human CD4, human CD28, or human CD8.

102. the transmembrane domain of said second CAR is or comprises a transmembrane domain from human CD28; and / or the transmembrane domain of the second CAR is or comprises the amino acid sequence set forth in SEQ ID NO: 18 or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to SEQ ID NO: 18; The polynucleotide of any one of claims 86 to 101.

103. The polynucleotide of any one of claims 86-102, wherein the intracellular signaling region of said second CAR comprises an intracellular signaling domain.

104. The polynucleotide of claim 103, wherein the intracellular signaling domain of said second CAR is or comprises the cytoplasmic signaling domain of the CD3-zeta (CD3ζ) chain or a functional variant or signaling portion thereof, optionally the cytoplasmic signaling domain of the human CD3 zeta chain.

105. 105. The polynucleotide of any one of claims 103 or 104, wherein the intracellular signaling region of the second CAR comprises the amino acid sequence set forth in SEQ ID NO:20, or an amino acid sequence having at least or at least about 90%, at least 91%, or at least about 91%, at least 92%, or at least about 92%, at least 93%, or at least about 93%, at least 94%, or at least about 94%, at least 95%, or at least about 95%, at least 96%, or at least about 96%, at least 97%, or at least about 97%, at least 98%, or at least about 98%, or at least 99% or at least about 99% sequence identity to SEQ ID NO:

20.

106. The polynucleotide of any one of claims 103-105, wherein the intracellular signaling region of said second CAR further comprises a costimulatory signaling region.

107. The polynucleotide of claim 106, wherein the costimulatory signaling region of said second CAR comprises the intracellular signaling domain of CD28, 4-1BB, or ICOS, or a signaling portion thereof, and optionally comprises the intracellular signaling domain of human CD28, human 4-1BB, or human ICOS.

108. 108. The polynucleotide of any one of claims 80-107, wherein at least one of said first chimeric antigen receptor and said second chimeric antigen receptor comprises an intracellular signaling region comprising the intracellular signaling domain of 4-1BB or a signaling portion thereof, optionally comprising the intracellular signaling domain of human 4-1BB.

109. The polynucleotide of any one of claims 106-108, wherein the costimulatory signaling region of said second CAR comprises the intracellular signaling domain of 4-1BB or a signaling portion thereof, optionally comprising the intracellular signaling domain of human 4-1BB.

110. the costimulatory signaling region of the second CAR comprises: the intracellular signaling domain of human CD28; and / or an amino acid sequence set forth in SEQ ID NO:46, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:46; 109. The polynucleotide of any one of claims 106 to 108, comprising:

111. the costimulatory signaling region of the second CAR comprises: the intracellular signaling domain of human 4-1BB; and / or an amino acid sequence set forth in SEQ ID NO:19, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:19; 110. The polynucleotide of any one of claims 106 to 109, comprising:

112. (i) a first nucleic acid sequence encoding a first chimeric antigen receptor (CAR) comprising a first antigen-binding domain; and (ii) a second nucleic acid sequence encoding a second chimeric antigen receptor (CAR) comprising a second antigen-binding domain; A polynucleotide comprising: the first CAR and the second CAR each comprise (a) the first antigen-binding domain or the second antigen-binding domain, (b) a spacer, (c) a transmembrane domain, and (d) an intracellular signaling region comprising an intracellular signaling domain and a costimulatory signaling region; one or more of (b)-(d) of the first CAR and the same one or more of (b)-(d) of the second CAR comprise the same amino acid sequence; and the nucleotide sequence encoding one or more of (b)-(d) of the first CAR differs in sequence from the nucleotide sequence encoding the same one or more of (b)-(d) of the second CAR; Polynucleotide.

113. (i) a first nucleic acid sequence encoding a first chimeric antigen receptor (CAR) comprising a first antigen-binding domain capable of binding to either GPRC5D or BCMA; and (ii) a second nucleic acid sequence encoding a second chimeric antigen receptor (CAR) comprising a second antigen-binding domain capable of binding to the other of GPRC5D or BCMA; A polynucleotide comprising: the first CAR and the second CAR each comprise (a) the first antigen-binding domain or the second antigen-binding domain, (b) a spacer, (c) a transmembrane domain, and (d) an intracellular signaling region comprising an intracellular signaling domain and a costimulatory signaling region; one or more of (b)-(d) of the first CAR and the same one or more of (b)-(d) of the second CAR comprise the same amino acid sequence; and the nucleotide sequence encoding one or more of (b)-(d) of the first CAR differs in sequence from the nucleotide sequence encoding the same one or more of (b)-(d) of the second CAR. Polynucleotide.

114. The polynucleotide of any one of claims 80 to 113, wherein at least one of the nucleotide sequence encoding the first chimeric antigen receptor and the nucleotide sequence encoding the second chimeric antigen receptor is of a codon-different form.

115. 1. A polynucleotide comprising: (i) a first nucleic acid sequence encoding a first chimeric antigen receptor (CAR); (ii) a second nucleic acid sequence encoding a second chimeric antigen receptor (CAR); and (iii) a nucleotide sequence encoding a multicistronic element, wherein the first nucleic acid encoding the first CAR and the second nucleic acid encoding the second CAR are separated by the multicistronic element; the first CAR comprises a first antigen-binding domain that binds to GPRC5D, optionally encoded by the nucleotide sequence set forth in SEQ ID NO: 311; a spacer encoded by the nucleotides set forth in SEQ ID NO: 305; a transmembrane domain encoded by the nucleotide sequence set forth in SEQ ID NO: 307; and an intracellular signaling region comprising an intracellular signaling domain encoded by the nucleotide sequence set forth in SEQ ID NO: 309 and a costimulatory signaling region encoded by the nucleotide sequence set forth in SEQ ID NO: 308; the second CAR comprises a second antigen-binding domain that binds to BCMA, optionally encoded by the nucleotide sequence set forth in SEQ ID NO:310; a spacer encoded by the nucleotides set forth in SEQ ID NO:74; a transmembrane domain encoded by the nucleotide sequence set forth in SEQ ID NO:56; and an intracellular signaling region comprising an intracellular signaling domain encoded by the nucleotide sequence set forth in SEQ ID NO:58 and a costimulatory signaling domain region encoded by the nucleotide sequence set forth in SEQ ID NO:60; a first nucleic acid sequence encoding the first CAR is located toward the 5' end of the polynucleotide relative to a second nucleic acid sequence encoding the second CAR; Polynucleotide.

116. 1. A polynucleotide comprising: (i) a first nucleic acid sequence encoding a first chimeric antigen receptor (CAR); (ii) a second nucleic acid sequence encoding a second chimeric antigen receptor (CAR); and (iii) a nucleotide sequence encoding a multicistronic element, wherein the first nucleic acid encoding the first CAR and the second nucleic acid encoding the second CAR are separated by the multicistronic element; the first CAR comprises a first antigen-binding domain that binds to BCMA, optionally encoded by the nucleotide sequence set forth in SEQ ID NO:310; a spacer encoded by the nucleotides set forth in SEQ ID NO:74; a transmembrane domain encoded by the nucleotide sequence set forth in SEQ ID NO:56; and an intracellular signaling region comprising an intracellular signaling domain encoded by the nucleotide sequence set forth in SEQ ID NO:58 and a costimulatory signaling domain region encoded by the nucleotide sequence set forth in SEQ ID NO:60; the second CAR comprises a second antigen-binding domain that binds to GPRC5D, optionally encoded by the nucleotide sequence set forth in SEQ ID NO: 311; a spacer encoded by the nucleotides set forth in SEQ ID NO: 305; a transmembrane domain encoded by the nucleotide sequence set forth in SEQ ID NO: 307; and an intracellular signaling region comprising an intracellular signaling domain encoded by the nucleotide sequence set forth in SEQ ID NO: 309 and a costimulatory signaling region encoded by the nucleotide sequence set forth in SEQ ID NO: 308; a first nucleic acid encoding the first CAR is located toward the 5' end of the polynucleotide relative to a second nucleic acid sequence encoding the second CAR; Polynucleotide.

117. 117. The polynucleotide of any one of claims 80-116, wherein the nucleotide sequence encoding the first chimeric antigen receptor and the nucleotide sequence encoding the second chimeric antigen receptor share no more than about 30, no more than about 20, or no more than about 10 contiguous base pairs of sequence homology.

118. A vector comprising the polynucleotide of any one of claims 68 to 117.

119. 119. The vector of claim 118, which is a viral vector.

120. 68. A cell comprising the chimeric antigen receptor of any one of claims 1 to 67.

121. 121. The cell of claim 120, wherein the chimeric antigen receptor is a first chimeric receptor, and the cell further comprises a polynucleotide comprising a nucleotide encoding a second chimeric antigen receptor.

122. A cell comprising the polynucleotide of any one of claims 68 to 117.

123. 80. A cell comprising the polynucleotide of any one of claims 68-79 which is a first polynucleotide, and further comprising a second polynucleotide comprising a nucleotide sequence encoding a second chimeric antigen receptor (CAR).

124. The cell of claim 121 or claim 123, wherein said second chimeric antigen receptor (CAR) comprises an extracellular antigen-binding domain that specifically binds to a second antigen expressed on or associated with multiple myeloma.

125. The cell of claim 124, wherein said second CAR further comprises a spacer, a transmembrane domain, and an intracellular signaling region.

126. The cell of claim 124 or claim 125, wherein the second antigen is selected from B-cell maturation antigen (BCMA), CD38, CD138, CS-1, BAFF-R, TACI, and FcRH5.

127. The cell of any one of claims 124 to 126, wherein the second antigen is BCMA.

128. The second CAR is (1) An extracellular antigen-binding domain that specifically binds to BCMA, (i) a heavy chain variable (V) comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:189, 191, 193, 195, or 197; H ) area; and (ii) a light chain variable (V) comprising an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:190, 192, 194, 196, or 198; L )region an extracellular antigen-binding domain comprising: (2) Spacer, optionally an IgG4 / 2 chimeric hinge or a modified IgG4 hinge; IgG2 / 4 chimeric C H 2 domains; and IgG4 C H a spacer comprising three regions and optionally being at or about 228 amino acids in length, or the spacer shown in SEQ ID NO: 17; (3) a transmembrane domain; and (4) Intracellular signal transduction region 128. The cell of any one of claims 121-127, comprising:

129. V of the second car H The region is V as set forth in SEQ ID NO: 189, 191, 193, 195, or 197. H CDR-H1, CDR-H2, and CDR-H3 contained within the V region amino acid sequence, L The region is selected from the group consisting of V, VL and VL sequences shown in SEQ ID NO: 190, 192, 194, 196, or 198. L The cell of claim 128, comprising CDR-L1, CDR-L2, and CDR-L3 contained within the region amino acid sequence.

130. The second CAR is (1) An extracellular antigen-binding domain that specifically binds to BCMA, (i) a heavy chain complementary determining region 1 (CDR-H1) comprising the amino acid sequence set forth in SEQ ID NO: 199, 202, 206, or 209; (b) a heavy chain complementary determining region 2 (CDR-H2) comprising the amino acid sequence set forth in SEQ ID NO: 200, 203, 207, or 210; and (c) a heavy chain complementary determining region 3 (CDR-H3) comprising the amino acid sequence set forth in SEQ ID NO: 201, 204, 205, 208, or 211. H ) area; and (ii) a light chain variable (V) CDR comprising: (i) a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 218, 221, 224, 227, or 230; (b) a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 219, 222, 225, 228, or 231; and (c) a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 220, 223, or 226. L )region an extracellular antigen-binding domain comprising: (2) Spacer, optionally an IgG4 / 2 chimeric hinge or a modified IgG4 hinge; IgG2 / 4 chimeric C H 2 domains; and IgG4 C H a spacer comprising three regions and optionally about 228 amino acids in length or the spacer set forth in SEQ ID NO: 17; (3) a transmembrane domain; and (4) Intracellular signal transduction region 128. The cell of any one of claims 121-127, comprising:

131. V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 199, 200, and 201, respectively, and L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 218, 219, and 220, respectively; V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 202, 203, and 204, respectively, and L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 221, 222, and 223, respectively; V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 199, 200, and 205, respectively, and L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 224, 225, and 226, respectively; V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 206, 207, and 208, respectively, and L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 227, 228, and 229, respectively; or V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and L The regions include CDR-L1, CDR-L2, and CDR-L3, each of which comprises the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; A cell according to any one of claims 128 to 130.

132. V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and L The cell of any one of claims 128 to 131, wherein the regions comprise CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively.

133. V of the second car H Area and V L a region having (a) the amino acid sequence set forth in SEQ ID NO:189 and SEQ ID NO:190, respectively; or (b) an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:189; and a region having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98% sequence identity to SEQ ID NO:

190. comprises an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to NO:190; V of the second car H Area and V L a region having (a) the amino acid sequence set forth in SEQ ID NO:191 and SEQ ID NO:192, respectively; or (b) an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:191; and a region having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:

192. or comprises an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to NO:192; V of the second car H Area and V L a region having (a) the amino acid sequence set forth in SEQ ID NO:193 and SEQ ID NO:194, respectively, or (b) an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:193, and comprises an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to NO:194; V of the second car H Area and V L a region having (a) the amino acid sequence set forth in SEQ ID NO:195 and SEQ ID NO:196, respectively, or (b) an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:195, and or comprises an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to NO:196; or V of the second car H Area and V L a region having (a) the amino acid sequence set forth in SEQ ID NO:197 and SEQ ID NO:198, respectively, or (b) an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:197, and comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to NO:198; A cell according to any one of claims 128 to 132.

134. V of the second car H Area and V L the regions comprise the amino acid sequences set forth in SEQ ID NO:189 and SEQ ID NO:190, respectively; V of the second car H Area and V L whether the region comprises the amino acid sequence set forth in SEQ ID NO:191 and SEQ ID NO:192; V of the second car H Area and V L whether the region comprises the amino acid sequence set forth in SEQ ID NO:193 and SEQ ID NO:194; V of the second car H Area and V L the region comprises the amino acid sequence set forth in SEQ ID NO:195 and SEQ ID NO:196; or V of the second car H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NO: 197 and SEQ ID NO: 198, respectively; A cell according to any one of claims 128 to 133.

135. The cell of claims 128-134, wherein the single-chain antibody fragment is or comprises a single-chain variable fragment (scFv).

136. V of the second car H The region is V of the second CAR L 136. The cell of any one of claims 128-135, wherein said region is amino terminal to said region.

137. V of the second car H The region is V of the second CAR L 136. The cell of any one of claims 128-135, wherein said region is carboxy-terminal to said region.

138. 138. The cell of any one of claims 128-137, wherein the antigen-binding domain of the second CAR comprises an amino acid sequence selected from SEQ ID NOs: 237, 238, 239, 240, and 241, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NOs: 237, 238, 239, 240, and 241.

139. The cell of any one of claims 128-138, wherein the antigen-binding domain comprises an amino acid sequence selected from SEQ ID NOs: 237, 238, 239, 240 and 241.

140. V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and L the regions comprise CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; and / or V of the second car H Area and V L the regions comprise the amino acid sequences set forth in SEQ ID NO:197 and SEQ ID NO:198, respectively; and / or the antigen-binding domain comprises the amino acid sequence set forth in SEQ ID NO:241, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:241; 140. The cell of any one of claims 128 to 139.

141. The cell of any one of claims 125-140, wherein the transmembrane domain of said second CAR is or comprises a transmembrane domain derived from CD4, CD28, or CD8, optionally a transmembrane domain derived from human CD4, human CD28, or human CD8.

142. The cell of any one of claims 125-141, wherein the intracellular signaling region of said second CAR further comprises a costimulatory signaling region.

143. The cell of claim 142, wherein said costimulatory signaling region comprises the intracellular signaling domain of CD28, 4-1BB, or ICOS, or a signaling portion thereof, optionally comprising the intracellular signaling domain of human CD28, human 4-1BB, or human ICOS.

144. The cell of claim 142 or claim 143, wherein said costimulatory signaling region comprises the intracellular signaling domain of 4-1BB or a signaling portion thereof, optionally comprising the intracellular signaling domain of human 4-1BB.

145. 68. A composition comprising the chimeric antigen receptor of any one of claims 1 to 67.

146. 145. A composition comprising a cell according to any one of claims 120-144 or a plurality of cells according to any one of claims 120-144.

147. The composition of claim 143, wherein said composition comprises CD4+ T cells and CD8+ T cells, and wherein the ratio of said CD4+ T cells to said CD8+ T cells is from about 1:3 to about 3:1, optionally from about 1:2 to about 2:

1.

148. a plurality of first cells comprising a first chimeric antigen receptor, wherein the first chimeric antigen receptor is the chimeric antigen receptor of any one of claims 1 to 67 or encoded by the polynucleotide of any one of claims 68 to 79; and a plurality of second cells comprising a second chimeric antigen receptor; A composition comprising:

149. The composition of claim 148, wherein the second chimeric receptor comprises an extracellular antigen-binding domain that specifically binds to a second antigen expressed on or associated with multiple myeloma.

150. The composition of claim 148 or claim 149, wherein the second CAR comprises an extracellular antigen-binding domain that binds to a second antigen, a spacer, a transmembrane domain, and an intracellular signaling region.

151. The composition of claim 149 or claim 150, wherein the second antigen is selected from B-cell maturation antigen (BCMA), CD38, CD138, CS-1, BAFF-R, TACI, and FcRH5.

152. The composition of any one of claims 149-151, wherein said second antigen is BCMA.

153. The second CAR is (1) An extracellular antigen-binding domain that specifically binds to BCMA, (i) a heavy chain variable (V) comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:189, 191, 193, 195, or 197; H ) area; and (ii) a light chain variable (V) comprising an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:190, 192, 194, 196, or 198; L )region an extracellular antigen-binding domain comprising: (2) Spacer, optionally an IgG4 / 2 chimeric hinge or a modified IgG4 hinge; IgG2 / 4 chimeric C H 2 domains; and IgG4 C H a spacer comprising three regions and optionally about 228 amino acids in length or the spacer set forth in SEQ ID NO: 17; (3) a transmembrane domain; and (4) Intracellular signal transduction region 153. The composition of any one of claims 148-152, comprising:

154. V of the second car H The region is V as set forth in SEQ ID NO: 189, 191, 193, 195, or 197. H CDR-H1, CDR-H2, and CDR-H3 contained within the V region amino acid sequence, L The region is selected from the group consisting of V, VL and VL sequences shown in SEQ ID NO: 190, 192, 194, 196, or 198. L The composition of claim 153, comprising CDR-L1, CDR-L2, and CDR-L3 contained within the region amino acid sequence.

155. The second CAR is (1) An extracellular antigen-binding domain that specifically binds to BCMA, (i) a heavy chain complementary determining region 1 (CDR-H1) comprising the amino acid sequence set forth in SEQ ID NO: 199, 202, 206, or 209; (b) a heavy chain complementary determining region 2 (CDR-H2) comprising the amino acid sequence set forth in SEQ ID NO: 200, 203, 207, or 210; and (c) a heavy chain complementary determining region 3 (CDR-H3) comprising the amino acid sequence set forth in SEQ ID NO: 201, 204, 205, 208, or 211. H ) area; and (ii) a light chain variable (V) CDR comprising: (i) a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 218, 221, 224, 227, or 230; (b) a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 219, 222, 225, 228, or 231; and (c) a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 220, 223, or 226. L )region an extracellular antigen-binding domain comprising: (2) Spacer, optionally an IgG4 / 2 chimeric hinge or a modified IgG4 hinge; IgG2 / 4 chimeric C H 2 domains; and IgG4 C H a spacer comprising three regions and optionally about 228 amino acids in length or the spacer set forth in SEQ ID NO: 17; (3) a transmembrane domain; and (4) Intracellular signal transduction region 153. The composition of any one of claims 148-152, comprising:

156. V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 199, 200, and 201, respectively, and L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 218, 219, and 220, respectively; V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 202, 203, and 204, respectively, and L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 221, 222, and 223, respectively; V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 199, 200, and 205, respectively, and L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 224, 225, and 226, respectively; V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 206, 207, and 208, respectively, and L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 227, 228, and 229, respectively; or V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and L The regions include CDR-L1, CDR-L2, and CDR-L3, each of which comprises the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; The composition of any one of claims 153-155.

157. V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and L The composition of any one of claims 153-156, wherein the regions comprise CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively.

158. V of the second car H Area and V L a region having (a) the amino acid sequence set forth in SEQ ID NO:189 and SEQ ID NO:190, respectively; or (b) an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:189; and a region having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98% sequence identity to SEQ ID NO:

190. comprises an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to NO:190; V of the second car H Area and V L a region having (a) the amino acid sequence set forth in SEQ ID NO:191 and SEQ ID NO:192, respectively; or (b) an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:191; and a region having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:

192. or comprises an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to NO:192; V of the second car H Area and V L a region having (a) the amino acid sequence set forth in SEQ ID NO:193 and SEQ ID NO:194, respectively, or (b) an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:193, and comprises an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to NO:194; V of the second car H Area and V L a region having (a) the amino acid sequence set forth in SEQ ID NO:195 and SEQ ID NO:196, respectively, or (b) an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:195, and or comprises an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to NO:196; or V of the second car H Area and V L a region having (a) the amino acid sequence set forth in SEQ ID NO:197 and SEQ ID NO:198, respectively, or (b) an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:197, and comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to NO:198; The composition of any one of claims 153-157.

159. V of the second car H Area and V L the regions comprise the amino acid sequences set forth in SEQ ID NO:189 and SEQ ID NO:190, respectively; V of the second car H Area and V L whether the region comprises the amino acid sequence set forth in SEQ ID NO:191 and SEQ ID NO:192; V of the second car H Area and V L whether the region comprises the amino acid sequence set forth in SEQ ID NO:193 and SEQ ID NO:194; V of the second car H Area and V L the region comprises the amino acid sequence set forth in SEQ ID NO:195 and SEQ ID NO:196; or V of the second car H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NO: 197 and SEQ ID NO: 198, respectively; The composition of any one of claims 153-158.

160. V of the second car H The region is V of the second CAR L The composition of any one of claims 153-159, wherein the composition is amino terminal to the region.

161. V of the second car H The region is V of the second CAR L The composition of any one of claims 153-159, wherein the composition is carboxy terminal to the region.

162. 162. The composition of any one of claims 153-161, wherein the antigen-binding domain of the second CAR comprises an amino acid sequence selected from SEQ ID NOs: 237, 238, 239, 240, and 241, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NOs: 237, 238, 239, 240, and 241.

163. The composition of any one of claims 153-162, wherein the antigen-binding domain of the second CAR comprises an amino acid sequence selected from SEQ ID NOs: 237, 238, 239, 240, and 241.

164. V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and L the regions comprise CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; and / or V of the second car H Area and V L the regions comprise the amino acid sequences set forth in SEQ ID NO:197 and SEQ ID NO:198, respectively; and / or the antigen-binding domain comprises the amino acid sequence set forth in SEQ ID NO:241, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:241; The composition of any one of claims 153-163.

165. The composition of any one of claims 150-164, wherein the transmembrane domain of said second CAR is or comprises a transmembrane domain derived from CD4, CD28, or CD8, optionally human CD4, human CD28, or human CD8.

166. the transmembrane domain of said second CAR is or comprises a transmembrane domain from human CD28; and / or the transmembrane domain of the second CAR is or comprises the amino acid sequence set forth in SEQ ID NO: 18 or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to SEQ ID NO: 18; The composition of any one of claims 150-165.

167. 167. The composition of any one of claims 150-166, wherein said intracellular signaling region comprises an intracellular signaling domain, wherein the intracellular signaling domain is capable of inducing a primary activation signal in a T cell, is a component of a T cell receptor (TCR), and / or comprises an immunoreceptor tyrosine-based activation motif (ITAM).

168. The composition of claim 167, wherein the intracellular signaling domain is or comprises the cytoplasmic signaling domain of the CD3-zeta (CD3ζ) chain or a functional variant or signaling portion thereof, optionally the cytoplasmic signaling domain of the human CD3 zeta chain.

169. The composition of any one of claims 150-168, wherein the intracellular signaling region of said second CAR further comprises a costimulatory signaling region.

170. The composition of claim 169, wherein the costimulatory signaling region of said second CAR comprises the intracellular signaling domain of CD28, 4-1BB, or ICOS, or a signaling portion thereof, and optionally comprises the intracellular signaling domain of human CD28, human 4-1BB, or human ICOS.

171. The composition of any one of claims 148-170, wherein at least one of said first chimeric antigen receptor and said second chimeric antigen receptor comprises an intracellular signaling region comprising the intracellular signaling domain of 4-1BB or a signaling portion thereof, optionally comprising the intracellular signaling domain of human 4-1BB.

172. The composition of any one of claims 169-171, wherein the costimulatory signaling region of said second CAR comprises the intracellular signaling domain of 4-1BB or a signaling portion thereof, optionally comprising the intracellular signaling domain of human 4-1BB.

173. 173. The composition of any one of claims 148-172, wherein said plurality of first cells comprises T cells, optionally comprising CD4+ T cells and CD8+ T cells, optionally at a ratio of said CD4+ T cells to said CD8+ T cells of about 1:3 to about 3:1, optionally 1:2 to 2:

1.

174. The composition of any one of claims 148-173, wherein said plurality of second cells comprises T cells, optionally comprising CD4+ T cells and CD8+ T cells, optionally at a ratio of said CD4+ T cells to said CD8+ T cells of about 1:3 to about 3:1, optionally about 1:2 to about 2:

1.

175. 175. The composition of any one of claims 148-174, wherein the ratio of said first plurality of cells to said second plurality of cells in said composition is from about 1:3 to about 3:1, optionally from about 1:2 to about 2:1, optionally about 1:

1.

176. 176. The composition of any one of claims 148-175, comprising said first plurality of cells expressing said first chimeric antigen receptor to said second plurality of cells expressing said second chimeric antigen receptor in a ratio that is from about 1:3 to about 3:1, optionally from about 1:2 to about 2:1, optionally about 1:

1.

177. 177. The pharmaceutical composition of any one of claims 145-176 for use in treating a subject having a disease or condition, optionally cancer.

178. 145. A pharmaceutical composition for use in the treatment of a disease or disorder, optionally cancer, comprising the cells of any one of claims 120 to 144 as an active ingredient.

179. 236. A pharmaceutical composition for use in the treatment of a disease or disorder, optionally cancer, comprising as an active ingredient the composition of any one of claims 145-176, 233 or 234.

180. 80. A pharmaceutical composition for use in the treatment of a disease or disorder, optionally cancer, comprising as active ingredients a composition comprising a first dose of a plurality of first cells comprising a first chimeric antigen receptor which is the chimeric antigen receptor of any one of claims 1 to 67 or encoded by the polynucleotide of any one of claims 68 to 79, and a composition comprising a second dose of a plurality of second cells comprising a second chimeric antigen receptor.

181. 13. A method of treatment comprising administering to a subject having a disease or disorder a dose of a composition comprising the cells of any of claims 120-144, or a composition of any of claims 145-180, 233 or 234.

182. 145. Use of the cells of any one of claims 120 to 144 for treating a disease or disorder, optionally cancer.

183. 236. Use of the composition of any one of claims 145-180, 233 or 234 for treating a disease or disorder, optionally cancer.

184. 145. Use of a cell according to any one of claims 120 to 144 for the manufacture of a medicament for treating a disease or disorder, optionally cancer.

185. 23. Use of the composition of any one of claims 145-180, 233 or 234 for the manufacture of a medicament for treating a disease or disorder, optionally cancer.

186. The dose of the cells is about 1.0 x 10 7 CAR-expressing T cells ~ approximately 1.2 x 10 9 CAR-expressing T cells, approximately 1.0 x 10 7 CAR-expressing T cells ~ approximately 6.5 x 10 8 CAR-expressing T cells, approximately 1.5 x 10 7 CAR-expressing T cells ~ approximately 6.5 x 10 8 CAR-expressing T cells, approximately 1.5 x 10 7 CAR-expressing T cells ~ approximately 6.0 x 10 8 CAR-expressing T cells, approximately 2.5 x 10 7 CAR-expressing T cells ~ approximately 6.0 x 10 8 CAR-expressing T cells, approximately 5.0 x 10 7 CAR-expressing T cells ~ approximately 6.0 x 10 8 CAR-expressing T cells, approximately 1.25 x 10 7 CAR-expressing T cells ~ approximately 1.2 x 10 9 CAR-expressing T cells, approximately 1.5 x 10 7 CAR-expressing T cells ~ approximately 1.2 x 10 9 CAR-expressing T cells, approximately 5.0 x 10 7 CAR-expressing T cells ~ approximately 4.5 x 10 8 CAR-expressing T cells, or approximately 1.5 x 10 8 CAR-expressing T cells ~ approximately 3.0 x 10 8 186. The method or use of any one of claims 181-185, or the pharmaceutical composition for use of any one of claims 177-180, comprising CAR-expressing T cells, each of the endpoints inclusive.

187. The cell dose is 1.5 x 10 7 Or about 1.5 x 10 7 pieces, 2.5×10 7 Or about 2.5 x 10 7 pieces, 5.0×10 7 or approximately 5.0 x 10 7 pieces, 7.5×10 7 Or about 7.5 x 10 7 pieces, 1.5×10 8 Or about 1.5 x 10 8 pieces, 2.25×10 8 Or about 2.25 x 10 8 pieces, 3.0×10 8 Or about 3.0 x 10 8 pieces, 4.5×10 8 or about 4.5 x 10 8 pieces, 6.0×10 8 or approximately 6.0 x 10 8 pieces, 8.0×10 8 or approximately 8.0 x 10 8 pieces, or 1.2 x 10 9 or about 1.2 x 10 9 187. The method or use of any one of claims 181-186 or the pharmaceutical composition for the use of any one of claims 177-180 comprising CAR-expressing T cells.

188. administering to a subject having a disease or disorder a composition comprising a first dose of a plurality of first cells comprising a first chimeric antigen receptor which is the chimeric antigen receptor of any one of claims 1-67 or encoded by the polynucleotide of any one of claims 68-79; and administering to the subject a composition comprising a second dose of a plurality of second cells comprising a second chimeric antigen receptor. A method of treatment comprising:

189. 80. Use of a composition comprising a first dose of a plurality of first cells comprising a first chimeric antigen receptor that is the chimeric antigen receptor of any one of claims 1 to 67 or encoded by the polynucleotide of any one of claims 68 to 79, and a composition comprising a second dose of a plurality of second cells that comprises a second chimeric antigen receptor, for the treatment of a disease or disorder, optionally wherein the disease or condition is cancer.

190. 80. Use of a composition comprising a first dose of a plurality of first cells comprising a first chimeric antigen receptor that is the chimeric antigen receptor of any one of claims 1 to 67 or encoded by the polynucleotide of any one of claims 68 to 79, and a composition comprising a second dose of a plurality of second cells that comprises a second chimeric antigen receptor, for the manufacture of a medicament for treating a disease or disorder, optionally wherein the disease or condition is cancer.

191. The first dose of the plurality of first cells and the second dose of the plurality of second cells are independently 1.0 x 10 7 or approximately 1.0 x 10 7 CAR-expressing T cells ~ 1.5 x 10 9 Or about 1.5 x 10 9 CAR-expressing T cells, 1.0 x 10 7 or approximately 1.0 x 10 7 CAR-expressing T cells ~ 6.5 x 10 8 or about 6.5 x 10 8 CAR-expressing T cells, 1.25 x 10 7 Or about 1.25 x 10 7 CAR-expressing T cells ~ 0.6 x 10 8 or approximately 0.6 x 10 8 CAR-expressing T cells, 1.5 x 10 7 Or about 1.5 x 10 7 CAR-expressing T cells ~ 6.5 x 10 8 or about 6.5 x 10 8 CAR-expressing T cells, 1.5 x 10 7 Or about 1.5 x 10 7 CAR-expressing T cells ~6.0 x 10 8 or approximately 6.0 x 10 8 CAR-expressing T cells, 2.5 x 10 7 Or about 2.5 x 10 7 CAR-expressing T cells ~2.25 x 10 8 Or about 2.25 x 10 8 CAR-expressing T cells, 2.5 x 10 7 Or about 2.5 x 10 7 CAR-expressing T cells ~6.0 x 10 8 or approximately 6.0 x 10 8 CAR-expressing T cells, 5.0 x 10 7 or approximately 5.0 x 10 7 CAR-expressing T cells ~6.0 x 10 8 or approximately 6.0 x 10 8 CAR-expressing T cells, 7.5 x 10 7 Or about 7.5 x 10 7 CAR-expressing T cells ~ 1.5 x 10 8 Or about 1.5 x 10 8 CAR-expressing T cells, 2.5 x 10 7 Or about 2.5 x 10 7 CAR-expressing T cells ~ 1.2 x 10 9 or about 1.2 x 10 9 CAR-expressing T cells, 5.0 x 10 7 or approximately 5.0 x 10 7 CAR-expressing T cells ~ 4.5 x 10 8 or about 4.5 x 10 8 CAR-expressing T cells, or 1.5 x 10 8 Or about 1.5 x 10 8 CAR-expressing T cells ~3.0 x 10 8 Or about 3.0 x 10 8 191. The method or use of any one of claims 188-190, comprising CAR-expressing T cells, each end point inclusive.

192. 191. The method or use of any one of claims 188-190, or the pharmaceutical composition for use of claim 180, wherein said second chimeric receptor comprises an extracellular antigen-binding domain that specifically binds to a second antigen expressed on or associated with multiple myeloma.

193. 193. The method or use or pharmaceutical composition for use of claim 192, wherein said second antigen is selected from B-cell maturation antigen (BCMA), CD38, CD138, CS-1, BAFF-R, TACI, and FcRH5.

194. 194. The method or use or pharmaceutical composition for use of any one of claims 188-193, wherein said second antigen is BCMA.

195. The second CAR is (1) An extracellular antigen-binding domain that specifically binds to BCMA, (i) a heavy chain variable (V) comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:189, 191, 193, 195, or 197; H ) area; and (ii) a light chain variable (V) comprising an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:190, 192, 194, 196, or 198; L )region an extracellular antigen-binding domain comprising: (2) Spacer, optionally an IgG4 / 2 chimeric hinge or a modified IgG4 hinge; IgG2 / 4 chimeric C H 2 domains; and IgG4 C H a spacer comprising three regions and optionally about 228 amino acids in length or the spacer set forth in SEQ ID NO: 17; (3) a transmembrane domain; and (4) Intracellular signal transduction region 195. The method or use or pharmaceutical composition for use of any one of claims 188-194, comprising:

196. V of the second car H The region is V as set forth in SEQ ID NO: 189, 191, 193, 195, or 197. H CDR-H1, CDR-H2, and CDR-H3 contained within the V region amino acid sequence, L The region is selected from the group consisting of V, VL and VL sequences shown in SEQ ID NO: 190, 192, 194, 196, or 198. L 196. The method or use or pharmaceutical composition for use of claim 195, comprising CDR-L1, CDR-L2, and CDR-L3 contained within the region amino acid sequence.

197. The second CAR is (1) An extracellular antigen-binding domain that specifically binds to BCMA, (i) a heavy chain complementary determining region 1 (CDR-H1) comprising an amino acid sequence selected from SEQ ID NOs: 199, 202, 206, and 209; (b) a heavy chain complementary determining region 2 (CDR-H2) comprising an amino acid sequence selected from SEQ ID NOs: 200, 203, 207, and 210; and (c) a heavy chain complementary determining region 3 (CDR-H3) comprising an amino acid sequence selected from SEQ ID NOs: 201, 204, 205, 208, and 211. H ) area; and (ii) a light chain variable (V) antibody comprising: (i) a light chain complementarity determining region 1 (CDR-L1) comprising an amino acid sequence selected from SEQ ID NOs: 218, 221, 224, 227, and 230; (b) a light chain complementarity determining region 2 (CDR-L2) comprising an amino acid sequence selected from SEQ ID NOs: 219, 222, 225, 228, and 231, 234; and (c) a light chain complementarity determining region 3 (CDR-L3) comprising an amino acid sequence selected from SEQ ID NOs: 220, 223, and 226. L )region an extracellular antigen-binding domain comprising: (2) Spacer, optionally an IgG4 / 2 chimeric hinge or a modified IgG4 hinge; IgG2 / 4 chimeric C H 2 domains; and IgG4 C H a spacer comprising three regions and optionally about 228 amino acids in length or the spacer set forth in SEQ ID NO: 17; (3) a transmembrane domain; and (4) Intracellular signal transduction region 197. The method or use or pharmaceutical composition for use of any one of claims 188-196, comprising:

198. V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 199, 200, and 201, respectively, and L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 218, 219, and 220, respectively; V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 202, 203, and 204, respectively, and L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 221, 222, and 223, respectively; V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 199, 200, and 205, respectively, and L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 224, 225, and 226, respectively; V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 206, 207, and 208, respectively, and L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 227, 228, and 229, respectively; or V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and L The regions include CDR-L1, CDR-L2, and CDR-L3, each of which comprises the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; 198. The method or use or pharmaceutical composition for use of any one of claims 195-197.

199. V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and L 199. The method or use or pharmaceutical composition for use of any one of claims 195-198, wherein said regions comprise CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 230, 231 and 232, respectively.

200. V of the second car H Area and V L a region having (a) the amino acid sequence set forth in SEQ ID NO:189 and SEQ ID NO:190, respectively; or (b) an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:189; and a region having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98% sequence identity to SEQ ID NO:

190. comprises an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to NO:190; V of the second car H Area and V L a region having (a) the amino acid sequence set forth in SEQ ID NO:191 and SEQ ID NO:192, respectively; or (b) an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:191; and a region having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:

192. or comprises an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to NO:192; V of the second car H Area and V L a region having (a) the amino acid sequence set forth in SEQ ID NO:193 and SEQ ID NO:194, respectively, or (b) an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:193, and comprises an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to NO:194; V of the second car H Area and V L a region having (a) the amino acid sequence set forth in SEQ ID NO:195 and SEQ ID NO:196, respectively, or (b) an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:195, and or comprises an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to NO:196; or V of the second car H Area and V L the region comprises (a) the amino acid sequence set forth in SEQ ID NO:197 and SEQ ID NO:198, respectively, or (b) an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to SEQ ID NO:197 and SEQ ID NO:198, 200. The method or use or pharmaceutical composition for use of any one of claims 195 to 199.

201. V of the second car H Area and V L the regions comprise the amino acid sequences set forth in SEQ ID NO:189 and SEQ ID NO:190, respectively; V of the second car H Area and V L whether the region comprises the amino acid sequence set forth in SEQ ID NO:191 and SEQ ID NO:192; V of the second car H Area and V L whether the region comprises the amino acid sequence set forth in SEQ ID NO:193 and SEQ ID NO:194; V of the second car H Area and V L the region comprises the amino acid sequence set forth in SEQ ID NO:195 and SEQ ID NO:196; or V of the second car H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NO: 197 and SEQ ID NO: 198, respectively; 201. The method or use or pharmaceutical composition for use of any one of claims 195-200.

202. V of the second car H The region V L 202. The method or use or pharmaceutical composition for use of any one of claims 195-201, wherein said region is carboxy terminal to said region.

203. 203. The method or use or pharmaceutical composition for use of any one of claims 195-202, wherein the antigen-binding domain of said second CAR comprises an amino acid sequence selected from SEQ ID NOs: 237, 238, 239, 240, and 241, or an amino acid sequence having at least or at least about 90%, at least 91%, or at least about 91%, at least 92%, or at least about 92%, at least 93%, or at least about 93%, at least 94%, or at least about 94%, at least 95%, or at least about 95%, at least 96%, or at least about 96%, at least 97%, or at least about 97%, at least 98%, or at least about 98%, or at least 99% or at least about 99% sequence identity to an amino acid sequence selected from SEQ ID NOs: 237, 238, 239, 240, and 241.

204. The method or use, or pharmaceutical composition for use, of any one of claims 195-203, wherein the antigen-binding domain of said second CAR comprises an amino acid sequence selected from SEQ ID NOs: 237, 238, 239, 240, and 241.

205. V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and L the regions comprise CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; and / or V of the second car H Area and V L the regions comprise the amino acid sequences set forth in SEQ ID NO:197 and SEQ ID NO:198, respectively; and / or the antigen-binding domain of the second CAR comprises the amino acid sequence set forth in SEQ ID NO:241, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:241; 205. The method or use or pharmaceutical composition for use of any one of claims 195-204.

206. The method or use or pharmaceutical composition for use of any one of claims 188-205, wherein at least one of said first chimeric antigen receptor and said second chimeric antigen receptor comprises an intracellular signaling region comprising the intracellular signaling domain of 4-1BB or a signaling portion thereof, optionally comprising the intracellular signaling domain of human 4-1BB.

207. The method or use or pharmaceutical composition for use of any one of claims 195-206, wherein the transmembrane domain of said second CAR is or comprises a transmembrane domain derived from CD4, CD28, or CD8, optionally a transmembrane domain derived from human CD4, human CD38, or human CD8.

208. the transmembrane domain of said second CAR is or comprises a transmembrane domain from human CD28; and / or the transmembrane domain of the second CAR is or comprises the amino acid sequence set forth in SEQ ID NO: 18 or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to SEQ ID NO: 18; 208. The method or use or pharmaceutical composition for use of any one of claims 195-207.

209. The method or use, or pharmaceutical composition for use, of any one of claims 195-208, wherein the intracellular signaling region of said second CAR comprises an intracellular signaling domain.

210. 210. The method or use or pharmaceutical composition for use of claim 209, wherein the intracellular signaling domain of said second CAR is or comprises the cytoplasmic signaling domain of the CD3-zeta (CD3ζ) chain or a functional variant or signaling portion thereof, optionally the cytoplasmic signaling domain of the human CD3 zeta chain.

211. 211. The method or use, or pharmaceutical composition for use, of claim 209 or 210, wherein the intracellular signaling region of said second CAR comprises the amino acid sequence set forth in SEQ ID NO:20, or an amino acid sequence having at least or at least about 90%, at least 91%, or at least about 91%, at least 92%, or at least about 92%, at least 93%, or at least about 93%, at least 94%, or at least about 94%, at least 95%, or at least about 95%, at least 96%, or at least about 96%, at least 97%, or at least about 97%, at least 98%, or at least about 98%, or at least 99% or at least about 99% sequence identity to SEQ ID NO:

20.

212. The method or use, or pharmaceutical composition for use, of any one of claims 209-211, wherein the intracellular signaling region of said second CAR further comprises a costimulatory signaling region.

213. 213. The method or use, or pharmaceutical composition for use, of claim 212, wherein the costimulatory signaling region of said second CAR comprises the intracellular signaling domain of CD28, 4-1BB, or ICOS, or a signaling portion thereof, and optionally comprises the intracellular signaling domain of human CD28, human 4-1BB, or human ICOS.

214. The method or use or pharmaceutical composition for use of claim 212 or claim 213, wherein the costimulatory signaling region of said second CAR comprises the intracellular signaling domain of 4-1BB or a signaling portion thereof, and optionally comprises the intracellular signaling domain of human 4-1BB.

215. the costimulatory signaling region of the second CAR comprises: the intracellular signaling domain of human CD28; and / or an amino acid sequence set forth in SEQ ID NO:46, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:46; 214. The method or use or pharmaceutical composition for use of claim 212 or claim 213, comprising:

216. the costimulatory signaling region of the second CAR comprises: the intracellular signaling domain of human 4-1BB; and / or an amino acid sequence set forth in SEQ ID NO:19, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:19; 215. The method or use or pharmaceutical composition for use of any one of claims 212-214, comprising:

217. 217. The method or use or pharmaceutical composition for use of any one of claims 181-216, wherein said disease or disorder is associated with expression of GPRC5D.

218. 218. The method or use or pharmaceutical composition for use of claim 217, wherein said disease or disorder is further associated with expression of B-cell maturation antigen (BCMA).

219. The method or use or pharmaceutical composition for use of any one of claims 181-218, wherein said disease or disorder is a B-cell related disorder.

220. The method or use or pharmaceutical composition for use of any one of claims 181-219, wherein said disease or disorder associated with BCMA is an autoimmune disease or disorder.

221. 221. The method or use or pharmaceutical composition for use of claim 220, wherein the autoimmune disease or disorder is systemic lupus erythematosus (SLE), lupus nephritis, inflammatory bowel disease, rheumatoid arthritis, ANCA-associated vasculitis, idiopathic thrombocytopenic purpura (ITP), thrombotic thrombocytopenic purpura (TTP), autoimmune thrombocytopenia, Chagas' disease, Graves' disease, Wegener's granulomatosis, polyarteritis nodosa, Sjogren's syndrome, pemphigus vulgaris, scleroderma, multiple sclerosis, psoriasis, IgA nephropathy, IgM polyneuropathy, vasculitis, diabetes mellitus, Raynaud's syndrome, antiphospholipid syndrome, Goodpasture's disease, Kawasaki's disease, autoimmune hemolytic anemia, myasthenia gravis, or progressive glomerulonephritis.

222. The method or use or pharmaceutical composition for use of any one of claims 181-221, wherein said disease or disorder is cancer.

223. 223. The method or use or pharmaceutical composition for use of claim 222, wherein the cancer is a GPRC5D-expressing cancer.

224. 224. The method or use or pharmaceutical composition for use of claim 222 or claim 223, wherein said cancer is a plasma cell malignancy, and said plasma cell malignancy is multiple myeloma (MM) or plasmacytoma.

225. 225. The method or use or pharmaceutical composition for use of any one of claims 222-224, wherein said cancer is multiple myeloma (MM).

226. 226. The method or use or pharmaceutical composition for use of claim 225, wherein said cancer is relapsed / refractory multiple myeloma.

227. the subject is refractory to BCMA-targeted therapy, optionally T cells comprising a CAR that specifically binds BCMA, or has relapsed after administration of BCMA-targeted therapy, optionally after administration of T cells comprising a CAR that specifically binds BCMA; or The method comprises selecting for treatment a subject who is refractory to a BCMA-targeted therapy, optionally T cells comprising a CAR that specifically binds BCMA, or who has relapsed after administration of a BCMA-targeted therapy, optionally after administration of T cells comprising a CAR that specifically binds BCMA.

227. The method or use or pharmaceutical composition for use of any one of claims 181-226.

228. The method or use, or pharmaceutical composition for use, of any one of claims 181-187 and 191-227, wherein said subject has previously undergone administration of a BCMA targeted therapy for the treatment of said disease or disorder prior to administration of said dose of cells.

229. The method or use or pharmaceutical composition for use of any one of claims 188-228, wherein said subject has previously undergone administration of a BCMA targeted therapy for the treatment of said disease or disorder prior to administration of said first dose of cells and said second dose of cells.

230. 230. The method or use or pharmaceutical composition for use of claim 228 or claim 229, wherein said BCMA targeted therapy comprises a composition comprising T cells comprising a CAR that specifically binds to BCMA.

231. 231. The method or use or pharmaceutical composition for use of any one of claims 228-230, wherein the subject is refractory to BCMA targeted therapy, optionally T cells comprising a CAR that specifically binds BCMA, or has relapsed after BCMA targeted therapy, optionally after administration of T cells comprising a CAR that specifically binds BCMA.

232. 232. The method or use or pharmaceutical composition for use of any one of claims 181-231, wherein said subject comprises multiple myeloma cells that exhibit loss of the BCMA antigen or epitope, downregulation of BCMA, and / or BCMA-negative tumor cells following the previous administration.

233. The composition of claim 146 or claim 147, wherein the composition comprises a plurality of cells, at least a portion of the cells comprising a first CAR that specifically binds to GPRC5D, and a portion of the cells comprising a second CAR that specifically binds to a second antigen expressed on or associated with multiple myeloma, optionally wherein the second antigen is BCMA, and a portion of the cells comprising both the first CAR and the second CAR.

234. The composition of claim 233, wherein the ratio of cells expressing the first CAR to cells expressing the second CAR is 1:3 to 3:1, or about 1:3 to about 3:1, optionally 1:2 to 2:1, or about 1:2 to about 2:1, optionally 1:1 or about 1:

1.

235. a plurality of first cells comprising a first chimeric antigen receptor which is the chimeric antigen receptor of any one of claims 1 to 67 and / or encoded by the polynucleotide of any one of claims 68 to 79; and a plurality of second cells comprising a second chimeric antigen receptor; Including, combinations.

236. 236. The combination of claim 235, wherein said second chimeric receptor comprises an extracellular antigen-binding domain that specifically binds to a second antigen expressed on or associated with multiple myeloma.

237. 237. The combination of claim 236, wherein said second antigen is selected from the group consisting of B-cell maturation antigen (BCMA), CD38, CD138, CS-1, BAFF-R, TACI, and FcRH5.

238. 238. The combination of claim 236 or claim 237, wherein said second antigen is BCMA.

239. The second CAR is (1) An extracellular antigen-binding domain that specifically binds to BCMA, (i) a heavy chain variable (V) comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NO:189, 191, 193, 195, or 197; H ) area; and (ii) a light chain variable (V) comprising an amino acid sequence having at least or at least about 90%, at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to the amino acid sequence set forth in any of SEQ ID NOs:190, 192, 194, 196, or 198. L )region an extracellular antigen-binding domain comprising: (2) Spacer, optionally an IgG4 / 2 chimeric hinge or a modified IgG4 hinge; IgG2 / 4 chimeric C H 2 domains; and IgG4 C H a spacer comprising three regions and optionally being about 228 amino acids in length, and / or a spacer set forth in SEQ ID NO: 17; (3) a transmembrane domain; and (4) Intracellular signal transduction region 239. The combination of any one of claims 235 to 238, comprising:

240. The above V H The region is V as set forth in SEQ ID NO: 189, 191, 193, 195, or 197. H CDR-H1, CDR-H2, and CDR-H3 contained within the V region amino acid sequence, L The region is selected from the group consisting of V, VL and VL sequences shown in SEQ ID NO: 190, 192, 194, 196, or 198. L 240. The combination of claim 239, comprising CDR-L1, CDR-L2, and CDR-L3 contained within the region amino acid sequence.

241. The second CAR is (1) An extracellular antigen-binding domain that specifically binds to BCMA, a heavy chain variable region (V) comprising: (i) a heavy chain complementarity determining region 1 (CDR-H1) comprising the amino acid sequence set forth in SEQ ID NO: 199, 202, 206, or 209; (b) a heavy chain complementarity determining region 2 (CDR-H2) comprising the amino acid sequence set forth in SEQ ID NO: 200, 203, 207, or 210; and (c) a heavy chain complementarity determining region 3 (CDR-H3) comprising the amino acid sequence set forth in SEQ ID NO: 201, 204, 205, 208, or 211. H ); and (ii) a light chain variable (V) CDR comprising: (i) a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 218, 221, 224, 227, 230, 233, or 235; (b) a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 219, 222, 225, 228, or 231; and (c) a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 220, 223, or 226. L )region an extracellular antigen-binding domain comprising: (2) Spacer, optionally an IgG4 / 2 chimeric hinge or a modified IgG4 hinge; IgG2 / 4 chimeric C H 2 domains; and IgG4 C H a spacer comprising three regions and optionally being about 228 amino acids in length, and / or a spacer set forth in SEQ ID NO: 17; (3) a transmembrane domain; and (4) Intracellular signal transduction region 241. The combination of any one of claims 235 to 240, comprising:

242. V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 199, 200, and 201, respectively, and L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 218, 219, and 220, respectively; V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 202, 203, and 204, respectively, and L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 221, 222, and 223, respectively; V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 199, 200, and 205, respectively, and L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 224, 225, and 226, respectively; V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 206, 207, and 208, respectively, and L the region comprises CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 227, 228, and 229, respectively; or V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and L The regions include CDR-L1, CDR-L2, and CDR-L3, each of which comprises the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; 242. A combination according to any one of claims 239 to 241.

243. V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and L 243. The combination of any one of claims 239 to 242, wherein the regions comprise CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively.

244. V of the second car H Area and V L a region having (a) the amino acid sequence set forth in SEQ ID NO:189 and SEQ ID NO:190, respectively; or (b) an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least about 99%, or 100% or about 100% sequence identity to SEQ ID NO:189; comprises an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to NO:190; V of the second car H Area and V L a region having (a) the amino acid sequence set forth in SEQ ID NO:191 and SEQ ID NO:192, respectively; or (b) an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:191; and a region having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:

192. or comprises an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to NO:192; V of the second car H Area and V L a region having (a) the amino acid sequence set forth in SEQ ID NO:193 and SEQ ID NO:194, respectively, or (b) an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:193, and comprises an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to NO:194; V of the second car H Area and V L a region having (a) the amino acid sequence set forth in SEQ ID NO:195 and SEQ ID NO:196, respectively, or (b) an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to SEQ ID NO:195, and or comprises an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to NO:196; or V of the second car H Area and V L a region having (a) the amino acid sequence set forth in SEQ ID NO:197 and SEQ ID NO:198, respectively, or (b) an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:197, and comprising an amino acid sequence having at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 98%, or at least 99% sequence identity to NO:198; 244. A combination according to any one of claims 240 to 243.

245. V of the second car H Area and V L the regions comprise the amino acid sequences set forth in SEQ ID NO:189 and SEQ ID NO:190, respectively; V of the second car H Area and V L whether the region comprises the amino acid sequence set forth in SEQ ID NO:191 and SEQ ID NO:192; V of the second car H Area and V L whether the region comprises the amino acid sequence set forth in SEQ ID NO:193 and SEQ ID NO:194; V of the second car H Area and V L the region comprises the amino acid sequence set forth in SEQ ID NO:195 and SEQ ID NO:196; or V of the second car H Area and V L The regions comprise the amino acid sequences set forth in SEQ ID NO: 197 and SEQ ID NO: 198, respectively; 245. The combination of any one of claims 239 to 244.

246. 246. The combination of any one of claims 239 to 245, wherein the antigen-binding domain comprises an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to the amino acid sequence set forth in SEQ ID NOs: 237, 238, 239, 240 or 241, or to the amino acid sequence set forth in SEQ ID NOs: 227, 238, 239, 240 or 241.

247. 247. The combination of any one of claims 239 to 246, wherein the antigen binding domain comprises the amino acid sequence shown in SEQ ID NO: 237, 238, 239, 240 or 241.

248. V of the second car H The second CAR region comprises CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 209, 210, and 211, respectively, and L the regions comprise CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of SEQ ID NOs: 230, 231, and 232, respectively; and / or V of the second car H the VL region and VL region comprise the amino acid sequences set forth in SEQ ID NO:197 and SEQ ID NO:198, respectively; and / or the antigen-binding domain comprises the amino acid sequence set forth in SEQ ID NO:241, or an amino acid sequence having at least or at least about 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least about 98%, or at least 99% sequence identity to the sequence set forth in SEQ ID NO:241; 248. The combination of any one of claims 239 to 247.

249. The combination of any one of claims 235-248, wherein at least one of said first chimeric antigen receptor and said second chimeric antigen receptor comprises an intracellular signaling region comprising the intracellular signaling domain of 4-1BB or a signaling portion thereof, optionally comprising the intracellular signaling domain of human 4-1BB.

250. 250. A kit comprising the combination of any one of claims 235-249 and instructions for use, optionally the instructions for administering a dose of a first plurality of cells and a second plurality of cells, optionally in accordance with the method or use or pharmaceutical composition for use of any one of claims 174-216.

251. An article of manufacture comprising the combination of any one of claims 235-249 or the kit of claim 250.

252. 252. The article of manufacture of claim 251, comprising a first container comprising a dose of said plurality of first cells and a second container comprising a dose of said plurality of second cells, optionally wherein said first container and said second container are independently vials or bags.

253. 250. Use of a combination according to any one of claims 235 to 249 for treating a disease or disorder, optionally cancer.

254. 250. Use of a combination according to any one of claims 235 to 249 for the manufacture of a medicament for the treatment of a disease or disorder, optionally cancer.

255. A pharmaceutical composition for treating a disease or disorder, optionally cancer, containing as an active ingredient a combination according to any one of claims 235 to 249.

Images