Therapeutic antibody epitope-tagged car t cells for enhanced control and safety

Abstract

The present disclosure relates generally to compositions and methods for improving the safety of adoptive immune cell therapy, including the use of a depletion tag (e.g., an epitope or a mimotope) that is capable of being bound by another molecule (e.g., an antibody) to facilitate the depletion of the engineered immune cells. One class of depletion tags disclosed herein contain an epitope or mimotope derived from a biomarker of a hematologic cancer (e.g., CCR4). A second class of depletion tags disclosed herein contain an epitope or mimotope derived from the same antigen that the engineered receptor binds to (e.g., a DLL3 derived depletion tag for engineered immune cells that bind to DLL3).

Description

THERAPEUTIC ANTIBODY EPITOPE-TAGGED CAR T CELLS FOR ENHANCED CONTROL AND SAFETYCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 572,671, filed on April 1, 2024, the contents of which are herein incorporated by reference in their entirety.STATEMENT REGARDING SEQUENCE LISTING

[0002] The Sequence Listing XML associated with this application is provided in XML file format and is hereby incorporated by reference into the specification. The name of the XML file containing the Sequence Listing XML is MNBI_003_01WO_SeqList_ST26.xml. The XML file is 665,052 bytes in size and created on March 27, 2025, and is being submitted electronically via USPTO Patent Center.FIELD

[0003] The present disclosure relates generally to compositions and methods for enhancing the control and safety of engineered T cell therapeutics. The disclosure provides recombinant receptors and engineered immune cells that contain one or more epitope or mimotope tags for a therapeutic antibody, such that, if an adverse event occurs the therapeutic antibody can be administered to remove the engineered immune cells and mitigate the adverse event. Also provided are recombinant nucleic acid constructs, vectors, particles and cells that comprise such recombinant nucleic acids, methods for preparing immune cells for use in cell therapies, as well as methods of mitigating adverse events in such cell therapies.BACKGROUND

[0004] Engineered immune cell therapies, such as adoptive T cell therapies using chimeric antigen receptor (CAR) T cells, have revolutionized cancer therapy. To date, five CD19 targeted CAR-T cell therapies have been approved by the FDA for use against hematological B cell cancers. Despite this success, meaningful responses have not been achieved for many tumors. Further, severe and sometimes life-threatening adverse events may be associated with engineered immune cell therapies.

[0005] One challenge for improving engineered immune cell thrapies is to ensure its safety while enhancing potency. The associated adverse events may arise due to an uncontrolled proliferation of the engineered cells in vivo, and thus depleting the engineered cells on demand is a promising strategy for improving the safety of such cell therapies. To date, a number of approaches have been proposed for depleting engineered immune cells, such as those employing a “safety switch” (e.g., a herpesvirus thymidine kinase (HSV-tk) gene, an Escherichia coli cytosine deaminase (EC-CD) gene, or an inducible Caspase9 (iCasp9) protein). However, the large size of such a separate polypeptide poses problems for AAV delivery of an expression cassette that encodes both the engineered receptor (e.g., CAR) and the separate depletion polypeptide. Further, these approaches often have limited depletion efficiency or relatively slow depletion rate in vivo. Thus, there remains significant unmet needs for improved strategies of depleting engineered immune cells to ensure the safety of corresponding cell therapies.SUMMARY

[0006] This section provides a general summary of the disclosure, and is not comprehensive of its full scope or all of its features.

[0007] In one aspect, the present disclosure provides a recombinant polypeptide comprising (i) an extracellular region comprising a binding region for an antigen; and (ii) a transmembrane region, wherein the extracellular region comprises a peptide epitope or mimotope capable of being bound by an anti-CCR4 antibody, wherein the recombinant polypeptide is not CCR4.

[0008] In some embodiments, the recombinant polypeptide is a chimeric antigen receptor (CAR) or a recombinant T cell receptor (TCR).

[0009] In some embodiments, the anti-CCR4 antibody is mogamulizumab, the peptide epitope consists of SEQ ID NO: 431, and the recombinant polypeptide does not comprise a cysteine immediately after the C-terminus of the peptide epitope or mimotope.

[0010] In some embodiments, the binding region comprises a heavy chain variable region (VH) and a light chain variable region (VL), and the peptide epitope or mimotope is located between the VH and the VL.

[0011] In one aspect, the present disclosure provides an engineered immune cell expressing a recombinant polypeptide described herein.

[0012] In one aspect, the present disclosure provides a method of treating cancer or an autoimmune disease in a subject in need thereof, comprising administering an effective amount of the engineered cells described herein to the subject. In some embodiments, the method further comprises administering the anti-CCR4 antibody to the subject, wherein the anti-CCR4 antibody is mogamulizumab. In some embodiments, the anti-CCR4 antibody is administered after detection or occurrence of an adverse event caused by the engineered cells in the subject, and wherein administration of the anti-CCR4 antibody reduces or eliminates at least one symptom of the adverse event. In some embodiments, the anti-CCR4 antibody depletes at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 99%, at least 99.5%, or at least 99.9% of the engineered cells in the subject.

[0013] In one aspect, the present disclosure provides modified peptide epitopes or mimotopes, wherein the peptide epitope or mimotope comprises or consists of an amino acid sequence having at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mutations (substitutions, insertions, and / or deletions) compared to an unmodified peptide epitope or mimotope, and wherein the modified peptide epitope or mimotope is capable of being bound by a monoclonal antibody.

[0014] In some embodiments, the peptide epitope comprises or consists of an amino acid sequence having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to SEQ ID NO: 430, and wherein the peptide epitope is capable of being bound by mogamulizumab. In some embodiments, the modified peptide epitope or mimotope does not comprise the cysteine that is present at the C-terminus of SEQ ID NO: 430. In some embodiments, the modified peptide epitope comprises or consists of any one of SEQ ID NO: 431-438. In some embodiments, the modified peptide epitope consists of SEQ ID NO: 431.

[0015] In some embodiments, the peptide epitope or mimotope comprises or consists of an amino acid sequence having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to SEQ ID NO: 475, and wherein the peptide epitope or mimotope is capable of being bound by rituximab. In some embodiments, the peptide epitope or mimotope comprises or consists of an amino acid sequence having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to SEQ ID NO: 476, and wherein the peptide epitope or mimotope is capable of being bound by Palivizumab. In some embodiments, the peptide epitope or mimotope comprises or consists of an amino acid sequenc having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to SEQ ID NO: 477, and wherein the peptide epitope or mimotope is capable of being bound by Cetuximab. In some embodiments, the peptide epitope or mimotopecomprises or consists of an amino acid sequence having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to SEQ ID NO: 478, and wherein the peptide epitope or mimotope is capable of being bound by Cetuximab. In some embodiments, the peptide epitope or mimotope comprises or consists of an amino acid sequence having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to SEQ ID NO: 479, and wherein the peptide epitope or mimotope is capable of being bound by Cetuximab. In some embodiments, the peptide epitope or mimotope comprises or consists of an amino acid sequence having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to SEQ ID NO: 480, and wherein the peptide epitope or mimotope is capable of being bound by Cetuximab. In some embodiments, the peptide epitope or mimotope comprises or consists of an amino acid sequence having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to SEQ ID NO: 481, and wherein the peptide epitope or mimotope is capable of being bound by Nivolumab. In some embodiments, the peptide epitope or mimotope comprises or consists of an amino acid sequence having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to SEQ ID NO: 482, and wherein the peptide epitope or mimotope is capable of being bound by Nivolumab. In some embodiments, the peptide epitope or mimotope comprises or consists of an amino acid sequence having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to SEQ ID NO: 483, and wherein the peptide epitope or mimotope is capable of being bound by QBend-10. In some embodiments, the peptide epitope or mimotope comprises or consists of an amino acid sequence having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to SEQ ID NO: 484, and wherein the peptide epitope or mimotope is capable of being bound by Alemtuzumab. In some embodiments, the peptide epitope or mimotope comprises or consists of an amino acid sequence having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to SEQ ID NO: 469, and wherein the peptide epitope or mimotope is capable of being bound by Sacituzumab. In some embodiments, the peptide epitope or mimotope comprises or consists of an amino acid sequence having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to any one of SEQ ID NO:470-474, and wherein the peptide epitope or mimotope is capable of being bound by an anti-GUCY2C antibody. In some embodiments, the peptide epitope or mimotope comprises or consists of an amino acid sequence having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / ordeletions) compared to SEQ ID NO: 439, and wherein the peptide epitope or mimotope is capable of being bound by Tarlatamab. In some embodiments, the peptide epitope or mimotope comprises or consists of an amino acid sequence having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to any one of SEQ ID NO: 441-468, and wherein the peptide epitope or mimotope is capable of being bound by an anti-DLL3 antibody. In some embodiments, the peptide epitope or mimotope comprises or consists of an amino acid sequence having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to any one of SEQ ID NOs: 519-524, and wherein the peptide epitope or mimotope is capable of being bound by to CAP-100.

[0016] In some embodiments, the mutation(s) are substitutions. In some embodiments, the mutation(s) are conservative substitutions.

[0017] In some embodiments, the cysteine(s) in the unmodified peptide epitope or mimotope are substituted or deleted. In some embodiments, the N-terminus of the unmodified peptide epitope or mimotope contains cysteine. In some embodiments, the C- terminus of the unmodified peptide epitope or mimotope contains cysteine. In some embodiments, the unmodified peptide epitope or mimotope comprises at least one cysteine between the N-terminus and the C-terminus. In some embodiments, the cysteine(s) in the unmodified peptide epitope or mimotope are substituted with serine, glycine, threonine, alanine, or valine. In some embodiments, the cysteine(s) in the unmodified peptide epitope or mimotope are substituted with serine or valine. In some embodiments, the cysteine(s) in the unmodified peptide epitope or mimotope are substituted with serine. In some embodiments, the cysteine(s) in the unmodified peptide epitope or mimotope are substituted with valine. In some embodiments, the modified peptide epitope or mimotope does not comprise any sulfur atoms. In some embodiments, the modified peptide epitope or mimotope does not comprise any cysteine(s).

[0018] In some embodiments, the mutation(s) comprise deleting or substituting one or more amino acids that facilitate polypeptide crosslinking in the unmodified peptide epitope or mimotope. In some embodiments, the mutation(s) comprise deleting or substituting one or more cysteine(s), lysine(s), aspartic acid(s), glutamic acid(s), or any combination thereof.

[0019] In some embodiments, the modified peptide epitope or mimotope is capable of being bound by the corresponding antibody with a dissociation constant (Kd) of less than 1 pM, less than 100 nM, less than 10 nM, or less than 1 nM. In some embodiments, the Kd ismeasured by surface plasmon resonance (SPR) using a sensor chip that contains the immobilized antibody.

[0020] In some embodiments, the modified peptide epitope or mimotope is capable of being bound by [fam-]trastuzumab deruxtecan, (fam-trastuzumab deruxtecan-nxki), Adebrelimab, Ado-trastuzumab emtansine, Alemtuzumab, Amivantamab, Atezolizumab, Avelumab, Belantamab mafodotin (belantamab mafodotin-blmf), Bevacizumab, Blinatumomab, Brentuximab vedotin, Cadonilimab, Camrelizumab, CAP- 100, Catumaxomab, Cemiplimab (cemiplimab-rwlc), Cetuximab, Cetuximab saratolacan, Cosibelimab, Daratumumab, Dinutuximab, Disitamab vedotin, Dostarlimab, Durvalumab, Edrecolomab, Elotuzumab, Elranatamab, Emapalumab (emapalumab-lzsg), Enfortumab vedotin (enfortumab vedotin-ejfv), Enlonstobart, Envafolimab, Epcoritamab, Gemtuzumab ozogamicin, Glofitamab, Ibritumomab tiuxetan, Inotuzumab ozogamicin, Ipilimumab, Isatuximab (isatuximab-irfc), Loncastuximab tesirine, Margetuximab-cmkb, Mirvetuximab soravtansine, Mogamulizumab (mogamulizumab-kpkc), Mosunetuzumab, Moxetumomab pasudotox (moxetumomab pasudotox-tdfk), Naxitamab-gqgk, Necitumumab, Nimotuzumab, Nivolumab, Cipterbin, Obinutuzumab, Ofatumumab, Olaratumab, Panitumumab, Pembrolizumab, Penpulimab, Pertuzumab, Polatuzumab vedotin (polatuzumab vedotin- piiq), Prolgolimab, Pucotenlimab, Racotumomab, Ramucirumab, Relatlimab, Retifanlimab, Ripertamab, Rituximab, Sacituzumab govitecan (sacituzumab govitecan-hziy), Serplulimab, Sintilimab, Socazolimab, Sugemalimab, Tafasitamab (tafasitamab-cxix), Tagitanlimab, Talquetamab (talquetamab-tgvs), Tebentafusp, Teclistamab, Tislelizumab, Tisotumab vedotin, tisotumab vedotin-tftv, Toripalimab, Tositumomab-1131, Trastuzumab, Trastuzumab duocarmazine, Tremelimumab, Zimberelimab, Zolbetuximab, Zuberitamab, Odronextamab, Ivonescimab, Benmelstobart, Trastuzumab botidotin, Iparomlimab, tuvonralimab, Tarlatamab, Patritumab deruxtecan, Sacituzumab tirumotecan, Zanidatamab, Linvoseltamab, Datopotamab deruxtecan, Anbenitamab, Anvatabart opadotin, Apamistamab-Iodine (1311), Bemarituzumab, Cetrelimab, Cobolimab, Domvanalimab, Emactuzumab, Erfonrilimab, Favezelimab, Felzartamab, Fianlimab, Finotonlimab, ABBV- 383, Geptanolimab, Gotistobart, Ivuxolimab, Izalontamab, Lemzoparlimab, Luveltamab tazevibulin, Magrolimab, Mecbotamab vedotin, Monalizumab, Nofazinlimab, Nurulimab, Ociperlimab, Oleclumab, Onfekafusp alfa, Oregovomab, Ozekibart, Pivekimab sunirine, Quavonlimab, Retlirafusp alfa, Rosopatamab (177Lu-DOTA), Rulonilimab, Sasanlimab, Telisotuzumab vedotin, Tiragolumab, IKS014, LCB14-0110, FS-1502, Trastuzumabmafodotin, Trastuzumab rezetecan, Vibostolimab, Vobramitamab duocarmazine, Zilovertamab vedotin, Suvemcitug, APX003, BD0801, TK001, sevacizumab, CTX-009, ES104, TR009, NOV1501, ABL001, DP303c, IAH0968, Kintuximab, gentuximab, cintuximab, MIL62, MRG002, MRG003, PM8002, MEDI5752, MCLA-128, AGEN1181, L19-IL2, L19-TNF, IMC-F106C, JMT101, BL-B01D1, BAT1308, AZD2936, JS004, TAB004, FG-M108, M108, 9MW2821, BMS-986349, CC-93269, EM801, ABBV-181, ABBV-151, ARGX-115, DS-7300a, IBI343, SHR-A1921, or ASKB589.

[0021] In some embodiments, the peptide epitope or mimotope comprises or consists of an amino acid sequence having 1, 2, 3, 4, or 5 mutations compared to the unmodified peptide epitope or mimotope.

[0022] In some embodiments, the unmodified peptide epitope or mimotope comprises or consists of no more than 10, no more than 15, no more than 20, no more than 25, no more than 30, no more than 35, no more than 40, no more than 45, or no more than 50, consecutive amino acids in an autoimmune-associated antigen or a cancer-associated antigen.

[0023] In one aspect, the present disclosure provides peptide epitopes comprising or consisting of SEQ ID NO: 430, wherein the peptide epitope is capable of being bound by an anti-CCR4 antibody. In some embodiments, the anti-CCR4 antibody is mogamulizumab.

[0024] In one aspect, the present disclosure provides peptide epitopes comprising or consisting of any one of SEQ ID NO: 439-468, wherein the peptide epitope is capable of being bound by an anti-DLL3 antibody. In some embodiments, the anti-DLL3 antibody is tarlatamab.

[0025] In one aspect, the present disclosure provides peptide epitopes comprising or consisting of any one of SEQ ID NO: 519-524, wherein the peptide epitope is capable of being bound by an anti-CCR7 antibody. In some embodiments, the anti-CCR7 antibody is CAP-100.

[0026] In one aspect, the present disclosure provides recombinant polypeptides comprising or consisting of the peptide epitope or mimotope of the disclosure. In some embodiments, the recombinant polypeptide consists of the peptide epitope or mimotope. In some embodiments, the peptide epitope or mimotope is expressed on the surface of a cell. In some embodiments, the peptide epitope or mimotope is located in an extracellular region of the recombinant polypeptide. In some embodiments, the recombinant polypeptide is not any of CCR4, CCR7, DLL3, TROP2, GUCY2C, CD20, RSV protein F, EGFR, PD-1, CD34, andCD52. In some embodiments, the recombinant polypeptide is not CCR4. In some embodiments, the recombinant polypeptide is not CCR7.

[0027] In some embodiments, the recombinant polypeptide does not comprise a cysteine at the C-terminus of the modified peptide epitope or mimotope. In some embodiments, the recombinant polypeptide does not comprise a cysteine immediately after the C-terminus of the modified peptide epitope or mimotope. In some embodiments, the recombinant polypeptide does not comprise a cysteine at the N-terminus of the modified peptide epitope or mimotope. In some embodiments, the recombinant polypeptide does not comprise a cysteine immediately before the N-terminus of the modified peptide epitope or mimotope.

[0028] In some embodiments, the recombinant polypeptide comprises one or more additional peptide epitope(s) or mimotope(s), wherein the one or more additional peptide epitope(s) or mimotope(s) are the same as the modified peptide epitope or different from the modified peptide epitope.

[0029] In one aspect, the present disclosure provides recombinant polypeptides comprising a tag, wherein the tag comprises or consists of a peptide epitope or a mimotope that mimics an epitope. In some embodiments, the epitope is derived from CCR4, DLL3, or CCR7.

[0030] In some embodiments, the recombinant polypeptide comprises a binding region for an antigen. In some embodiments, the antigen is expressed on the surface of a target cell.

[0031] In some embodiments, the recombinant polypeptide does not comprise a chimeric antigen receptor (CAR) or a recombinant T cell receptor (TCR).

[0032] In some embodiments, the recombinant polypeptide comprises or consists of a T cell receptor fusion construct (TRuC) derived from a CD3 protein and comprising the binding region. In some embodiments, the CD3 protein is CD3s.

[0033] In some embodiments, the recombinant polypeptide comprises or consists of a chimeric antigen receptor (CAR) or a recombinant T cell receptor (TCR).

[0034] In some embodiments, the recombinant polypeptide comprises or consists of a receptor comprising: (a) a heavy chain variable region (VH) located before the N-terminus of a constant region of TCRalpha, and a light chain variable region (VL) located before the N-terminus of a constant region of TCRbeta; or (b) a heavy chain variable region (VH) located before the N-terminus of a constant region of TCRbeta, and a light chain variable region (VL) located before the N-terminus of a constant region of TCRalpha, optionally, wherein at least one of the constant region of TCRalpha or the constant region of TCRbetaare an endogenous constant region of TCRalpha or an endogenous constant region of TCRbeta, optionally wherein the receptor is a T-body.

[0035] In some embodiments, the peptide epitope or mimotope is located between the VH and the constant region of TCRalpha. In some embodiments, the peptide epitope or mimotope is located between the VL and the constant region of TCRalpha. In some embodiments, the peptide epitope or mimotope is located between the VH and the constant region of TCRbeta. In some embodiments, the peptide epitope or mimotope is located between the VL and the constant region of TCRbeta. In some embodiments, the peptide epitope or mimotope is located N-terminal to the VH. In some embodiments, the peptide epitope or mimotope is located N-terminal to the VL. In some embodiments, the peptide epitope or mimotope is located C-terminal to the constant region of TCRalpha. In some embodiments, the peptide epitope or mimotope is located C-terminal to the constant region of TCRbeta.

[0036] In some embodiments, an immune cell expressing the recombinant polypeptide exhibits one or more of the following: (a) improved binding to the antigen expressed on the surface of the target cell; (b) reduced recombinant polypeptide -induced antigen-independent signaling and / or activation of the immune cell; and / or (c) reduced cross-linking between the recombinant polypeptides expressed on the cell surface of the immune cell; compared to an immune cell expressing a recombinant polypeptide comprising the unmodified peptide epitope or mimotope.

[0037] In some embodiments, an immune cell expressing the recombinant polypeptide exhibits one or more of the following: (a) comparable levels of binding to the antigen expressed on the surface of the target cell; (b) comparable levels of CAR-induced or TCR- induced antigen-independent signaling and / or activation of the immune cell; and / or (c) comparable levels of cross-linking between the recombinant polypeptides expressed on the cell surface of the immune cell; compared to an immune cell expressing a recombinant polypeptide that does not comprise a peptide epitope or mimotope.

[0038] In some embodiments, the recombinant polypeptide comprises the peptide epitope or mimotope located N-terminal to the binding region. In some embodiments, the peptide epitope or mimotope is separated from the N-terminus of the binding region by no more than 15, no more than 10, no more than 5, no more than 4, no more than 3, no more than 2, no more than 1, or zero amino acid(s).

[0039] In some embodiments, the recombinant polypeptide comprises the peptide epitope or mimotope located C-terminal to the binding region. In some embodiments, the peptide epitope or mimotope is separated from the C-terminus of the binding region by zero, no more than 5, no more than 10, no more than 15, no more than 20, no more than 25, no more than 30, no more than 35, no more than 40, no more than 45, no more than 50, no more than 55, or no more than 60 amino acids.

[0040] In some embodiments, the peptide epitope or mimotope is separated from the binding region by no more than 10, no more than 15, no more than 20, no more than 25, no more than 30, no more than 35, no more than 40, no more than 45, or no more than 50 amino acids.

[0041] In some embodiments, the peptide epitope or mimotope is located within the binding region.

[0042] In some embodiments, the TCR is TCRalpha, and the binding region comprises a variable region and a constant region. In some embodiments, the TCR is TCRbeta, and the binding region comprises a variable region and a constant region. In some embodiments, the peptide epitope or mimotope is located between the variable region and the constant region. In some embodiments, the peptide epitope or mimotope is located N-terminal to the variable region. In some embodiments, the peptide epitope or mimotope is located C-terminal to the constant region.

[0043] In some embodiments, the recombinant polypeptide comprises or consists of the CAR.

[0044] In some embodiments, the binding region comprises a heavy chain variable domain (VH) and a light chain variable domain (VL). In some embodiments, the binding region comprises a single chain variable fragment (scFv) comprising a VH and a VL. In some embodiments, the VH and the VL is separated by a linker.

[0045] In some embodiments, the binding region comprises one or more variable heavy domains of heavy chain (VHHs). In some embodiments, the binding region comprises at least two VHHs separated by a linker.

[0046] In some embodiments, the binding region comprises a first binding domain and a second binding domain, wherein the first binding domain and the second binding domain are separated by a linker.

[0047] In some embodiments, the recombinant polypeptide comprises a second binding region located N-terminal or C-terminal to the binding region. In some embodiments, the binding region and the second binding region are separated by a linker.

[0048] In some embodiments, the peptide epitope or mimotope is located within the linker.

[0049] In some embodiments, the linker comprises no more than 10, no more than 15, no more than 20, no more than 25, no more than 30, no more than 35, no more than 40, no more than 45, or no more than 50 amino acids.

[0050] In some embodiments, the peptide epitope or mimotope is located in between the binding region(s) and a transmembrane region. In some embodiments, the peptide epitope or mimotope is located within a hinge region that connects the binding region(s) to a transmembrane region. In some embodiments, the hinge region comprises no more than 10, no more than 15, no more than 20, no more than 25, no more than 30, no more than 35, no more than 40, no more than 45, no more than 50, no more than 55, no more than 60, no more than 65, or no more than 70 amino acids.

[0051] In some embodiments, the CAR or TCR comprises no more than 10, no more than 15, no more than 20, no more than 25, no more than 30, no more than 35, no more than 40, no more than 45, or no more than 50 amino acids N-terminal to the binding region(s), excluding the signal peptide.

[0052] In some embodiments, the extracellular region of the CAR or TCR comprises no more than 200, no more than 250, no more than 300, no more than 350, no more than 400, no more than 450, or no more than 500 amino acids.

[0053] In some embodiments, the CAR or TCR comprises, from N-terminus to C- terminus, the binding region(s), a hinge region, a transmembrane region.

[0054] In some embodiments, the antigen is an autoimmune-associated antigen or a cancer-associated antigen. In some embodiments, the peptide epitope is derived from an autoimmune-associated antigen or a cancer-associated antigen or wherein the mimotope mimics an epitope of the autoimmune-associated antigen or the cancer-associated antigen.

[0055] In some embodiments, the autoimmune-associated antigen or the cancer-associated antigen is selected from the group consisting of CD1, CD la, CD lb, CDlc, CD Id, CDle, CD2, CD3d, CD3e, CD3g, CD3s, CD4, CD5, CD7, CD8a, CD8b, CD19, CD20, CD21, CD22, CD23, CD24, CD25, CD27, CD28, CD30, CD33, CD34, CD38, CD40, CD44v6, CD45, CD46, CD47 CD48, CD52, CD59, CD66, CD70, CD71, CD72, CD73, CD79A, CD79B, CD80 (B7.1), CD86 (B7.2), CD94, CD95, CD97, CD123, CD134, CD140(PDGFR4), CD152, CD154, CD158, CD171, CD178, CD179, CD179a, CD181 (CXCR1), CD182 (CXCR2), CD183 (CXCR3), CD210, CD213A2, CD246, CD252, CD253, CD261, CD262, CD272, CD273 (PD-L2), CD274 (PD-L1), CD276 (B7H3), CD279, CD295, CD339 (JAG1), CD340 (HER2), CDH17, CEA, CLECL1, CLL-1, CLDN6, CLDN18.2, CS1, DLL3, LY6G6D, GCC, p53R175H, PRAME, EGFR, EGFRvIII, FGFR2, AFP, CA125, MUC-1, MAGE, ALPI, alkaline phosphatase placental-like 2 (ALPPL2), B-cell maturation antigen (BCM A), green fluorescent protein (GFP), enhanced green fluorescent protein (eGFP), KLK2, KLK3, Mesothelin, IL13Ra2, signal regulatory protein a (SIRPa), TCRalpha, TCRbeta, TSHR, GD2, GD3, Tn Ag, cMET, Axl, R0R1, R0R2, GPC1, GPC2, GPC3, FLT3, TAG72, CEA, EPC AM, KIT (CD117), IL-13Ra2, IL-l lRa, PSCA, PRSS21, VEGFR2, LewisY, PDGFRp, SSEA-4, folate receptor alpha, ERBB2 (Her2 / neu), MUC1, MUC16, NCAM, prostase, PAP, ELF2M, Ephrin B2, IGF-I receptor, CAIX, LMP2, gplOO, bcr-abl, tyrosinase, EphA2, STEAP1, STEAP2, fucosyl GM1, sLe, GM3, TGS5, HMWMAA, o-acetyl-GD2, folate receptor beta, TEM1 / CD248, TEM7R, R0PN1, GPRC5D, GPA33, CX0RF61, ALK, Polysialic acid, PLAC1, GloboH, NY-BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP, WT1, NY-ESO-1, LAGE-la, MAGE- Al, legumain, HPV E6,E7, MAGE A4, ETV6-AML, sperm protein 17, XAGE1, Tie 2, MAD-CT-1, MAD-CT-2, Fos-related antigen 1, p53, p53 mutant, p53R175H, KRAS, mutant KRAS, KRAS G12D, prostein, surviving, telomerase, PCTA-l / Galectin 8, MelanA / MARTl, Ras mutant, hTERT, sarcoma translocation breakpoints, ML-IAP, ERG (TMPRSS2 ETS fusion gene), NA17, PAX3, androgen receptor, cyclin Bl, MYCN, RhoC, TRP-2, CYP1B1, BORIS, SART3, PAX5, OY-TES1, LCK, AKAP-4, SSX2, RAGE-1, human telomerase reverse transcriptase, RU1, RU2, intestinal carboxyl esterase, mut hsp70- 2,LAIR1, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, FCRL5, IGLL1, PSMA, TROP2, citrullinated vimentin, the extracellular portion of the APRIL protein, and any combinations thereof.

[0056] In some embodiments, the cancer-associated antigen is a cancer-associated antigen for a solid tumor. In some embodiments, the cancer is a solid tumor selected from the group consisting of small cell lung cancer, prostate cancer, colorectal cancer, head and neck squamous cell carcinoma, breast cancer, melanoma, ovarian cancer, lung cancer, pancreatic cancer, stomach cancer, cervical cancer, hepatocellular carcinoma, glioma cancer, neuroblastoma cancer, and renal cell carcinoma.

[0057] In some embodiments, the cancer-associated antigen for the solid tumor comprises or consists of KIR2D, NRP1, CD221, VEGFR, HGF, integrin a5 1, CD51, CD27, CDH17, B7H3, NYESO1, EGFR, cMET, BRD4, EpCAM, SSX, Fibronectin EDB, integrin al 1 1, DLL3, CD2, SSTR2, TROP2, KLK2, KLK3, STEAP2, STEAP1, PSMA, PSCA, LY6G6D, IL-la, CEA, C242 antigen, CanAg (MUC1 glycoform), CTAA16.88, UCY2C, IGF2BP3, p53R175H, NKG2D ligands, PRAME, MAGEA4, vimentin, HER2 / neu, CCR4, CCR5, ROPN1, GPA33, ROR1, KK-LC-1, CA-125, folate receptor 1, folate receptor alpha, MUC1, MUC16, MSLN, CLDN6, WT1, ALPPL2, KRASG12D, KRASG12V, CD70, MSLN, CLDN18.2, HPV E6 / E7, GPC3, IL-13Ra, EGFRv3, EGFR806, GD2, or CA9. In some embodiments, the cancer-associated antigen for the solid tumor comprises or consists of KIR2D, NRP1, CD221, VEGFR, HGF, integrin a5bl, CD51, CD27, B7H3, NYESO1, EGFR, cMET, BRD4, EpCAM, SSX, Fibronectin EDB, or Integrin al 101. In some embodiments, the cancer-associated antigen for the solid tumor comprises or consists of KLK2, Ly6G6D, CLDN6, DLL3 or CD 19.

[0058] In some embodiments, the cancer is a hematologic cancer selected from B cell lymphoma, T cell lymphoma, chronic lymphocytic leukemia, and acute myeloid leukemia.

[0059] In some embodiments, the cancer-associated antigen is or comprises an antigen selected from Table 2. In some embodiments, the antigen is not CD19. In some embodiments, the antigen is not CCR4. In some embodiments, the antigen is not CCR7. In some embodiments, the antigen is DLL3.

[0060] In some embodiments, the epitope is derived from a cancer-associated antigen selected from Table 2. In some embodiments, the epitope is derived from DLL3.

[0061] In some embodiments, the peptide epitope comprises or consists of at least 10, at least 15, at least 20, at least 25, at least 30, at least 35, or at least 40 consecutive amino acids in SEQ ID NO: 486, or an amino acid sequence having at most 1, at most 2, at most 3, at most 4, or at most 5 amino acid mutations thereto. In some embodiments, the peptide epitope comprises or consists of any one of SEQ ID NO: 439-468, or an amino acid sequence having at most 1, at most 2, at most 3, at most 4, or at most 5 amino acid mutations thereto. In some embodiments, the peptide epitope comprises or consists of SEQ ID NO: 439, or an amino acid sequence having at most 1, at most 2, at most 3, at most 4, or at most 5 amino acid mutations thereto.

[0062] In some embodiments, the peptide epitope or mimotope is capable of being bound by a monoclonal antibody that binds to the antigen for the binding region. In someembodiments, the epitope is derived from a cancer-associated antigen of T cell lymphoma. In some embodiments, the epitope is derived from a cancer-associated antigen of adult T- cell lymphoma (ATL) and / or cutaneous T-cell lymphoma (CTCL).

[0063] In some embodiments, the epitope is derived from a cancer-associated antigen of a Th2 cell, a cutaneous lymphocyte antigen-positive skin-homing T cell, and / or a Treg cell.

[0064] In some embodiments, the epitope is derived from CCR4. In some embodiments, the peptide epitope comprises or consists of an amino acid sequence of any one of SEQ ID NO: 430-438, or an amino acid sequence having at most 1, at most 2, at most 3, at most 4, or at most 5 amino acid mutations thereto. In some embodiments, the peptide epitope comprises or consists of an amino acid sequence of SEQ ID NO: 431.

[0065] In some embodiments, the peptide epitope is derived from CCR7. In some embodiments, the peptide epitope comprises or consists of an amino acid sequence of any one of SEQ ID NOs: 519-524, or an amino acid sequence having at most 1, at most 2, at most 3, at most 4, or at most 5 amino acid mutations thereto. In some embodiments, the peptide epitope comprises or consists of an amino acid sequence of SEQ ID NOs: 519-524.

[0066] In some embodiments, the peptide epitope or the mimotope is about 5 to about 10, about 10 to about 20, about 15 to about 25, about 20 to about 30, about 25 to about 35, about 30 to about 40, about 35 to about 45, or about 40 to about 50 amino acids in length. In some embodiments, the peptide epitope or the mimotope is no more than 10, no more than 15, no more than 20, no more than 25, no more than 30, no more than 35, no more than 40, no more than 45, or no more than 50 amino acids in length.

[0067] In some embodiments, the peptide epitope does not comprise any mutation compared to the native epitope derived from the antigen.

[0068] In some embodiments, the peptide epitope or the mimotope is capable of forming an alpha-helical conformation.

[0069] In some embodiments, the peptide epitope or the mimotope is capable of being bound by an antibody with a dissociation constant (Kd) of less than 1 pM, less than 100 nM, less than 10 nM, or less than 1 nM. In some embodiments, the Kd is measured by surface plasmon resonance (SPR) using a sensor chip that contains the immobilized antibody. In some embodiments, the antibody is [fam-]trastuzumab deruxtecan, (fam-trastuzumab deruxtecan-nxki), Adebrelimab, Ado-trastuzumab emtansine, Alemtuzumab, Amivantamab, Atezolizumab, Avelumab, Belantamab mafodotin (belantamab mafodotin-blmf), Bevacizumab, Blinatumomab, Brentuximab vedotin, Cadonilimab, Camrelizumab, CAP-100, Catumaxomab, Cemiplimab (cemiplimab-rwlc), Cetuximab, Cetuximab saratolacan, Cosibelimab, Daratumumab, Dinutuximab, Disitamab vedotin, Dostarlimab, Durvalumab, Edrecolomab, Elotuzumab, Elranatamab, Emapalumab (emapalumab-lzsg), Enfortumab vedotin (enfortumab vedotin-ejfv), Enlonstobart, Envafolimab, Epcoritamab, Gemtuzumab ozogamicin, Glofitamab, Ibritumomab tiuxetan, Inotuzumab ozogamicin, Ipilimumab, Isatuximab (isatuximab-irfc), Loncastuximab tesirine, Margetuximab-cmkb, Mirvetuximab soravtansine, Mogamulizumab (mogamulizumab-kpkc), Mosunetuzumab, Moxetumomab pasudotox (moxetumomab pasudotox-tdfk), Naxitamab-gqgk, Necitumumab, Nimotuzumab, Nivolumab, Cipterbin, Obinutuzumab, Ofatumumab, Olaratumab, Panitumumab, Pembrolizumab, Penpulimab, Pertuzumab, Polatuzumab vedotin (polatuzumab vedotin- piiq), Prolgolimab, Pucotenlimab, Racotumomab, Ramucirumab, Relatlimab, Retifanlimab, Ripertamab, Rituximab, Sacituzumab govitecan (sacituzumab govitecan-hziy), Serplulimab, Sintilimab, Socazolimab, Sugemalimab, Tafasitamab (tafasitamab-cxix), Tagitanlimab, Talquetamab (talquetamab-tgvs), Tebentafusp, Teclistamab, Tislelizumab, Tisotumab vedotin, tisotumab vedotin-tftv, Toripalimab, Tositumomab-1131, Trastuzumab, Trastuzumab duocarmazine, Tremelimumab, Zimberelimab, Zolbetuximab, Zuberitamab, Odronextamab, Ivonescimab, Benmelstobart, Trastuzumab botidotin, Iparomlimab, tuvonralimab, Tarlatamab, Patritumab deruxtecan, Sacituzumab tirumotecan, Zanidatamab, Linvoseltamab, Datopotamab deruxtecan, Anbenitamab, Anvatabart opadotin, Apamistamab-Iodine (1311), Bemarituzumab, Cetrelimab, Cobolimab, Domvanalimab, Emactuzumab, Erfonrilimab, Favezelimab, Felzartamab, Fianlimab, Finotonlimab, ABBV- 383, Geptanolimab, Gotistobart, Ivuxolimab, Izalontamab, Lemzoparlimab, Luveltamab tazevibulin, Magrolimab, Mecbotamab vedotin, Monalizumab, Nofazinlimab, Nurulimab, Ociperlimab, Oleclumab, Onfekafusp alfa, Oregovomab, Ozekibart, Pivekimab sunirine, Quavonlimab, Retlirafusp alfa, Rosopatamab (177Lu-DOTA), Rulonilimab, Sasanlimab, Telisotuzumab vedotin, Tiragolumab, IKS014, LCB14-0110, FS-1502, Trastuzumab mafodotin, Trastuzumab rezetecan, Vibostolimab, Vobramitamab duocarmazine, Zilovertamab vedotin, Suvemcitug, APX003, BD0801, TK001, sevacizumab, CTX-009, ES104, TR009, NOV1501, ABL001, DP303c, IAH0968, Kintuximab, gentuximab, cintuximab, MIL62, MRG002, MRG003, PM8002, MEDI5752, MCLA-128, AGEN1181, L19-IL2, L19-TNF, IMC-F106C, JMT101, BL-B01D1, BAT1308, AZD2936, JS004, TAB004, FG-M108, M108, 9MW2821, BMS-986349, CC-93269, EM801, ABBV-181, ABBV-151, ARGX-115, DS-7300a, IBI343, SHR-A1921, or ASKB589 .

[0070] In some embodiments, the antibody is a bi-specific or multi-specific antibody that is capable of binding to (i) the peptide epitope or the mimotope; and (ii) CD3. In some embodiments, the antibody is a bi-specific or multi-specific antibody that is capable of specifically binding to (i) the peptide epitope or the mimotope; and (ii) CD 16. In some embodiments, the antibody is a tri-specific or multi-specific antibody that is capable of specifically binding to (i) the peptide epitope or the mimotope; (ii) 4 IBB or albumin, and (ii) CD3. In some embodiments, the antibody is tarlatamab.

[0071] In some embodiments, the antibody is mogamulizumab.

[0072] In some embodiments, the antibody is CAP- 100.

[0073] In some embodiments, the recombinant polypeptide comprises one or more additional peptide epitope(s) or mimotope(s). In some embodiments, the one or more additional peptide epitope(s) or mimotope(s) are the same as the peptide epitope or mimotope in the tag. In some embodiments, the one or more additional peptide epitope(s) or mimotope(s) are different from the peptide epitope or mimotope in the tag. In some embodiments, the recombinant polypeptide comprises 1, 2, 3, or 4 additional peptide epitope(s) or mimotope(s).

[0074] In some embodiments, the recombinant polypeptide comprises an extracellular portion comprising the binding region and the peptide epitope or mimotope.

[0075] In some embodiments, the recombinant polypeptide comprises a transmembrane region. In some embodiments, the recombinant polypeptide comprises a glycosylphosphatidylinositol (GPI) anchor.

[0076] In some embodiments, the recombinant polypeptide comprises an intracellular region. In some embodiments, the intracellular domain is located C-terminal to the transmembrane region. In some embodiments, the intracellular region comprises one or more intracellular signaling domains. In some embodiments, the one or more intracellular signaling domains are derived from a protein selected from the group consisting of CD3zeta, lxxCD3zeta, CD28, 4- IBB, 0X40, IL2Rb turbodomains, MYD88 / CD40, MYD88 / CD40 inducible costimulatory domain, CD2, B7-1 / CD80; B7-2 / CD86; B7-H1 / PD-L1; B7-H2; B7- H3; B7-H4; B7-H6; B7-H7; BTLA / CD272; CD28; CTLA-4; Gi24 / VISTA / B7-H5;ICOS / CD278; PD-1; PD-L2 / B7-DC; PDCD6); 4-1BB / TNFSF9 / CD137; 4-1BB Ligand / TNFSF9; BAFF / BLyS / TNFSF13B; BAFF R / TNFRSF13C; CD27 / TNFRSF7; CD27 Ligand / TNFSF7; CD30 / TNFRSF8; CD30 Ligand / TNFSF8; CD40 / TNFRSF5;CD40 / TNFSF5; CD40 Ligand / TNFSF5; DR3 / TNFRSF25; GITR / TNFRSF18; GITRLigand / TNFSF18; HVEM / TNFRSF14; LIGHT / TNFSF14; Lymphotoxin-alpha / TNF-beta; OX40 / TNFRSF4; 0X40 Ligand / TNFSF4; RELT / TNFRSF19L; TACI / TNFRSF13B; TL1A / TNFSF15; TNF-alpha; TNF RII / TNFRSF1B); 2B4 / CD244 / SLAMF4; BLAME / SLAMF8; CD2; CD2F-10 / SLAMF9; CD48 / SLAMF2; CD58 / LFA-3; CD84 / SLAMF5; CD229 / SLAMF3; CRACC / SLAMF7; NTB- A / SLAMF6; SLAM / CD150); CD2; CD7; CD53; CD82 / Kai-1; CD90 / Thyl; CD96; CD160; CD200; CD300a / LMIRl; HLA Class I; HLA-DR; Ikaros; Integrin alpha 4 / CD49d; Integrin alpha 4 beta 1; Integrin alpha 4 beta 7 / LPAM-l; LAG-3; TCL1A; TCL1B; CRTAM; DAP12; Dectin- 1 / CLEC7A; DPPIV / CD26; EphB6; TIM- 1 / KIM- 1 / HA VCR; TIM-4; TSLP; TSLP R; lymphocyte function associated antigen- 1 (LFA-1); NKG2C, an immunoreceptor tyrosine based activation motif (ITAM), CD27, CD134 / OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD7, LIGHT, NKG2C, and any combinations thereof. In some embodiments, the intracellular region comprises an intracellular signaling domain derived from 4-1BB and / or an intracellular signaling domain derived from CD3zeta. In some embodiments, the intracellular region comprises, from N-term to C-term, the intracellular signaling domain derived from 4-1BB and the intracellular signaling domain derived from CD3zeta. In some embodiments, the intracellular region comprises the intracellular signaling domain derived from 4-1BB comprising a polypeptide sequence at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 422. In some embodiments, the intracellular region comprises the intracellular signaling domain derived from CD3zeta comprising a polypeptide sequence at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 423. In some embodiments, the intracellular region comprises the intracellular signaling domain derived from CD3zeta comprising a polypeptide sequence at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 488 wherein the amino acid corresponding to position 14 of SEQ ID NO: 488 is lysine.

[0077] In some embodiments, the transmembrane region comprises a polypeptide sequence at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 421.

[0078] In some embodiments, the hinge region comprises a polypeptide sequence at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 420.

[0079] In some embodiments, the linker comprises a polypeptide sequence at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical to any one of SEQ ID NO: 416-418.

[0080] In some embodiments, the recombinant polypeptide comprises a polypeptide sequence at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical to any one of SEQ ID NO: 401-413.

[0081] In some embodiments, the recombinant polypeptide is no more than 200, no more than 250, no more than 300, no more than 350, no more than 400, no more than 450, no more than 500, no more than 550, no more than 600, no more than 650, no more than 700, no more than 750, or no more than 800 amino acids in length.

[0082] In one aspect, the present disclosure provides recombinant nucleic acids encoding the peptide epitope or mimotope of the disclosure.

[0083] In one aspect, the present disclosure provides recombinant nucleic acids encoding the recombinant polypeptide of the disclosure.

[0084] In some embodiments, the recombinant nucleic acid encodes the TCRalpha comprising the peptide epitope or mimotope and a TCRbeta. In some embodiments, the recombinant nucleic acid encodes a TCRalpha and the TCRbeta comprising the peptide epitope or mimotope.

[0085] In some embodiments, the recombinant nucleic acid encodes a TCRalpha, a TCRbeta, and the peptide epitope or mimotope located in between the TCRalpha and the TCRbeta.

[0086] In some embodiments, the TCRalpha and the TCRbeta are encoded by the same coding region. In some embodiments, the TCRalpha and the TCRbeta are separated by a cleavage site.

[0087] In some embodiments, the recombinant nucleic acid encodes (i) the VH and the constant region of TCRalpha of the receptor, or (ii) the VL and the constant region of TCRbeta of the receptor, wherein (i) or (ii) comprises the peptide epitope or mimotope. In some embodiments, the recombinant nucleic acid encodes (i) the VH and the constant region of TCRbeta of the receptor, or (ii) the VL and the constant region of TCRalpha of the receptor, wherein (i) or (ii) comprises the peptide epitope or mimotope. In some embodiments, (i) and (ii) are encoded by the same coding region; optionally, wherein (i) and (ii) are separated by a cleavage site.

[0088] In one aspect, the present disclosure provides recombinant nucleic acids encoding (i) a recombinant receptor that comprises a binding region for an antigen, and (ii) a recombinant polypeptide comprising a tag in its extracellular region, wherein the tag comprises a peptide epitope or a mimotope that mimics an epitope.

[0089] In some embodiments, the recombinant nucleic acid is less than 5.0 kb in length. In some embodiments, the recombinant nucleic acid is less than 4.8 kb in length. In some embodiments, the recombinant nucleic acid is less than 4.7 kb in length.

[0090] In some embodiments, the recombinant nucleic acid further comprises a polynucleotide sequence encoding a second recombinant polypeptide comprising a caspase- associated recruitment domain (CARD) containing protein or a functional fragment thereof. In some embodiments, the CARD containing protein comprises (i) a CARD domain comprising a sequence having at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to amino acids 11-103 of SEQ ID NO: 1 and (ii) a Src Homology region 2 (SH2) domain comprising a sequence having at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to amino acids 622-716 of SEQ ID NO: 1. In some embodiments, the CARD containing protein comprises a sequence having at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to any one of SEQ ID NO: 1-4. In some embodiments, the second recombinant polypeptide comprising the CARD containing protein or the functional fragment thereof comprises no more than 500, no more than 550, no more than 600, no more than 650, or no more than 700 amino acids.

[0091] In some embodiments, the polynucleotide sequence encoding the recombinant polypeptide is operably linked to a promoter. In some embodiments, the promoter is or comprises a minimal TATA promoter, pGK promoter, actin promoter, CD4 promoter, CD8a promoter, CD8b promoter, TCRa promoter, TCRb promoter, CD3d promoter, CD3g promoter, CD3e promoter, CD3z promoter, CARD9 promoter, CARDIO promoter, CARD 11 promoter, CARD 14 promoter, PIK3R3 promoter, CD25 promoter, IL2 promoter, IL7 promoter, IL 15 promoter, KLRG-1 promoter, HLA-DR promoter, CD38 promoter, CD69 promoter, Ki-67 promoter, CDl la promoter, CD58 promoter, CD99 promoter, CD62L promoter, CD 103 promoter, CCR4 promoter, CCR5 promoter, CCR6 promoter, CCR9 promoter, CCR10 promoter, CXCR3 promoter, CXCR4 promoter, CL A promoter, Granzyme A promoter, Granzyme B promoter, Perforin promoter, CD57 promoter, CD161 promoter, IL-18Ra promoter, CD69 promoter, GzmB promoter, T-bet promoter, IFNgammapromoter, TIM3 promoter, IL4 promoter, GAT A3 promoter, IL1 promoter, IL5 promoter, IL6 promoter, IL 13 promoter, IL 10 promoter, IL17A promoter, IL6 promoter, IL21 promoter, IL23R promoter, FoxP3 promoter, CTLA4 promoter, CD25 promoter, PD1 promoter, CD45RO promoter, CCR7 promoter, CD28 promoter, CD95 promoter, CD28 promoter, CD27 promoter, CD 127 promoter, PD-1 promoter, CD 122 promoter, CD 132 promoter, c-Kit promoter, nuclear factor of activated T cells (NF AT) promoter, programmed death 1 (PD-1) promoter, T cell immunoglobulin mucin-3 (TIM-3) promoter, cytotoxic T lymphocyte antigen-4 (CTLA4) promoter, lymphocyte-activation protein 3 (LAG-3) promoter, tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) promoter, B- and T-lymphocyte attenuator (BTLA) promoter, CD25 promoter, CD69 promoter, Fas ligand (FasL) promoter, TIGIT promoter, TGF-beta promoter, T-bet promoter, Eomes promoter, GATA3 promoter, CD45RA promoter, 2B4 promoter, Type I interferon (IFN) alpha, Type I IFN beta promoter, IFN gamma promoter, IRF3 promoter, IRF7 promoter, NFkB promoter, AP-1 promoter, TNF-alpha promoter, CD 130 promoter, NR4A1 promoter, NR4A2, NR4A3 promoter, MND promoter, EF-1 alpha promoter, short EF-1 alpha promoter, CAG promoter, ubiquitin / S27a promoter, SV40 promoter, SV40 early promoter, adenovirus major late promoter, mouse metallothionein-I promoter, Moloney murine leukemia virus (MMLV) long terminal repeat (LTR) region, CMV promoter, immunoglobulin promoter, heat shock promoter, polyoma virus promoter, fowlpox virus promoter, bovine papilloma virus promoter, avian sarcoma virus promoter, retrovirus promoter, hepatitis-B virus promoter, PGK promoter, vaccinia virus 7.5K promoter, TK promoter of HSV, mouse mammary tumor virus (MMTV) promoter, LTR promoter of HIV, promoter of moloney virus, Epstein Barr virus (EBV) promoter, Rous sarcoma virus (RSV) promoter, U6 promoter, or UBC promoter. In some embodiments, the promoter is a constitutive promoter. In some embodiments, the constitutive promoter is or comprises a CD4 promoter, CD8a promoter, CD8b promoter, TCRa promoter, TCRb promoter, CD3d promoter, CD3g promoter, CD3e promoter, CD3z promoter, CARD9 promoter, CARDIO promoter, CARD 11 promoter, CARD 14 promoter, PIK3R3 promoter, MND promoter or a short EFla promoter.

[0092] In some embodiments, the polynucleotide sequence encoding the recombinant polypeptide is not operably linked to a promoter.

[0093] In some embodiments, the recombinant nucleic acid further comprises a polynucleotide sequence encoding one or more cleavable linkers. In some embodiments, the one or more cleavable linkers comprise a P2A, E2A, F2A, or T2A self-cleaving peptide.

[0094] In some embodiments, the recombinant nucleic acid further comprises one or more nucleic acid element sequences. In some embodiments, wherein the one or more nucleic acid element sequences is or comprises ribosomal binding sites, enhancer elements, activator elements, translational start sequences, translational termination sequences, transcription start sequences, transcription termination sequences, polyadenylation signal sequences, a 70 bp poly(A) tract, a 100 bp poly(A) tract, a 172 bp poly(A) tract, a 200 bp poly(A) tract, a 300 bp poly(A) tract, a 325 bp poly(A) tract, replication elements, RNA processing and export elements, transposon sequences, transposase sequences, insulator sequences, internal ribosome entry sites (IRES), 5’UTRs, 3’UTRs, mRNA 3’ end processing sequences, boundary elements, locus control regions (LCR), matrix attachment regions (MAR), recombination or cassette exchange sequences, linker sequences, cleavable linker sequences, secretion signals, resistance markers, anchoring peptides, localization signals, fusion tags, affinity tags, chaperonins, proteases, or any combination thereof.

[0095] In some embodiments, the recombinant nucleic acid further comprises a polynucleotide sequence encoding one or more additional polypeptides and / or a non-coding RNA. In some embodiments, the non-coding RNA comprises a shRNA or a microRNA.

[0096] In some embodiments, the one or more additional polypeptides comprise an additional potency enhancement polypeptide, a cytokine, a chemokine, a growth factor, or any combination thereof. In some embodiments, the additional potency enhancement polypeptide is or comprises a dominant negative form of an inhibitor of a cell-mediated immune response of the immune cell (e.g., TGFPR2 DNR), c-Jun, CCL19, CCL21, IL2R, IL7, IL7Ralpha, IL15, IL15RA, IL18, decoy-resistant IL 18 (DR-18), MyD88 / CD40, PD1- CD28 switch receptor, PD1-41BB switch receptor, CD40L-CD28 switch receptor, CTBR12 switch receptor, or CD8alpha / beta.

[0097] In some embodiments, the recombinant nucleic acid is less than 10 kb, less than 9 kb, less than 8 kb, less than 7 kb, less than 6 kb, less than 5 kb, less than 4 kb, or less than 3 kb in length. In some embodiments, the recombinant nucleic acid is less than about 5.0 kb in length. In some embodiments, the recombinant nucleic acid is less than about 4.7 kb in length.

[0098] In one aspect, the present disclosure provides recombinant nucleic acids comprising, from 5’ to 3’: (1) a promoter, (2) a polynucleotide sequence encoding a CARD- containing protein or a functional fragment thereof, (3) a cleavable linker encoding sequence, and(4) a polynucleotide sequence encoding the recombinant polypeptide of the disclosure.

[0099] In one aspect, the present disclosure provides recombinant nucleic acids comprising, from 5’ to 3’ : (1) a promoter, (2) a polynucleotide sequence encoding the recombinant polypeptide of the disclosure, (3) a cleavable linker encoding sequence, and(4) a polynucleotide sequence encoding a CARD-containing protein or a functional fragment thereof.

[0100] In one aspect, the present disclosure provides recombinant nucleic acids comprising, from 5’ to 3’: (1) a promoter, (2) a polynucleotide sequence encoding a CARD- containing protein or a functional fragment thereof, (3) a cleavable linker encoding sequence, (4) a polynucleotide sequence encoding the recombinant polypeptide of the disclosure,(5) a cleavable linker encoding sequence, and (6) a polynucleotide sequence encoding an additional potency enhancement polypeptide, a cytokine, a chemokine, or a growth factor.

[0101] In one aspect, the present disclosure provides recombinant nucleic acids comprising, from 5’ to 3’ : (1) a promoter, (2) a polynucleotide sequence encoding the recombinant polypeptide of the disclosure, (3) a cleavable linker encoding sequence,(4) a polynucleotide sequence encoding a CARD-containing protein or a functional fragment thereof, (5) a cleavable linker encoding sequence, and (6) a polynucleotide sequence encoding an additional potency enhancement polypeptide, a cytokine, a chemokine, or a growth factor.

[0102] In one aspect, the present disclosure provides recombinant nucleic acids comprising, from 5’ to 3’ : (1) a promoter, (2) a polynucleotide sequence encoding the recombinant polypeptide of the disclosure, (3) a cleavable linker encoding sequence, (4) a polynucleotide sequence encoding an additional potency enhancement polypeptide, a cytokine, a chemokine, or a growth factor; (5) a cleavable linker encoding sequence, and(6) a polynucleotide sequence encoding a CARD-containing protein or a functional fragment thereof.

[0103] In one aspect, the present disclosure provides recombinant nucleic acids comprising, from 5’ to 3’ : (1) a promoter, (2) a polynucleotide sequence encoding anadditional potency enhancement polypeptide, a cytokine, a chemokine, or a growth factor, (3) a cleavable linker encoding sequence, (4) a polynucleotide sequence encoding the recombinant polypeptide of the disclosure, (5) a cleavable linker encoding sequence, and (6) a polynucleotide sequence encoding a CARD-containing protein or a functional fragment thereof.

[0104] In one aspect, the present disclosure provides recombinant nucleic acids comprising, from 5’ to 3’ : (1) a promoter, (2) a polynucleotide sequence encoding an additional potency enhancement polypeptide, a cytokine, a chemokine, or a growth factor, (3) a cleavable linker encoding sequence, (4) a polynucleotide sequence encoding a CARD- containing protein or a functional fragment thereof, (5) a cleavable linker encoding sequence, and (6) a polynucleotide sequence encoding the recombinant polypeptide of the disclosure.

[0105] In one aspect, the present disclosure provides recombinant nucleic acids comprising, from 5’ to 3’ : (1) a cleavable linker encoding sequence, (2) a polynucleotide sequence encoding a CARD-containing protein or a functional fragment thereof, (3) a cleavable linker encoding sequence, (4) a polynucleotide sequence encoding the recombinant polypeptide of the disclosure, and (5) a cleavable linker encoding sequence.

[0106] In one aspect, the present disclosure provides recombinant nucleic acids comprising, from 5’ to 3’ : (1) a cleavable linker encoding sequence, (2) a polynucleotide sequence encoding the recombinant polypeptide of the disclosure, (3) a cleavable linker encoding sequence, (4) a polynucleotide sequence encoding a CARD-containing protein or a functional fragment thereof, and (5) a cleavable linker encoding sequence.

[0107] In one aspect, the present disclosure provides recombinant nucleic acids comprising, from 5’ to 3’ : (1) a cleavable linker encoding sequence, (2) a polynucleotide sequence encoding a CARD-containing protein or a functional fragment thereof, (3) a cleavable linker encoding sequence, (4) a polynucleotide sequence encoding the recombinant polypeptide of the disclosure, (5) a cleavable linker encoding sequence, (6) a polynucleotide sequence encoding an additional potency enhancement polypeptide, a cytokine, a chemokine, or a growth factor, and (7) a cleavable linker encoding sequence.

[0108] In one aspect, the present disclosure provides recombinant nucleic acids comprising, from 5’ to 3’ : (1) a cleavable linker encoding sequence, (2) a polynucleotide sequence encoding a CARD-containing protein or a functional fragment thereof, (3) a cleavable linker encoding sequence, (4) a polynucleotide sequence encoding an additionalpotency enhancement polypeptide, a cytokine, a chemokine, or a growth factor, (5) a cleavable linker encoding sequence, (6) a polynucleotide sequence encoding the recombinant polypeptide of the disclosure, (7) a cleavable linker encoding sequence.

[0109] In one aspect, the present disclosure provides recombinant nucleic acids comprising, from 5’ to 3’ : (1) a cleavable linker encoding sequence, (2) a polynucleotide sequence encoding the recombinant polypeptide of the disclosure, (3) a cleavable linker encoding sequence, (4) a polynucleotide sequence encoding a CARD-containing protein or a functional fragment thereof, (5) a cleavable linker encoding sequence, (6) a polynucleotide sequence encoding an additional potency enhancement polypeptide, a cytokine, a chemokine, or a growth factor, and (7) a cleavable linker encoding sequence.

[0110] In one aspect, the present disclosure provides recombinant nucleic acids comprising, from 5’ to 3’ : (1) a cleavable linker encoding sequence, (2) a polynucleotide sequence encoding the recombinant polypeptide of the disclosure, (3) a cleavable linker encoding sequence, (4) a polynucleotide sequence encoding an additional potency enhancement polypeptide, a cytokine, a chemokine, or a growth factor, (5) a cleavable linker encoding sequence, (6) a polynucleotide sequence encoding a CARD-containing protein or a functional fragment thereof, and (7) a cleavable linker encoding sequence.[OHl] In one aspect, the present disclosure provides recombinant nucleic acids comprising, from 5’ to 3’ : (1) a cleavable linker encoding sequence, (2) a polynucleotide sequence encoding an additional potency enhancement polypeptide, a cytokine, a chemokine, or a growth factor,(3) a cleavable linker encoding sequence, (4) a polynucleotide sequence encoding a CARD- containing protein or a functional fragment thereof, (5) a cleavable linker encoding sequence,(6) a polynucleotide sequence encoding the recombinant polypeptide of the disclosure, and(7) a cleavable linker encoding sequence.

[0112] In one aspect, the present disclosure provides recombinant nucleic acids comprising, from 5’ to 3’ : (1) a cleavable linker encoding sequence, (2) a polynucleotide sequence encoding an additional potency enhancement polypeptide, a cytokine, a chemokine, or a growth factor, (3) a cleavable linker encoding sequence, (4) a polynucleotide sequence encoding the recombinant polypeptide of the disclosure, (5) a cleavable linker encoding sequence, (6) a polynucleotide sequence encoding a CARD-containing protein or a functional fragment thereof, and (7) a cleavable linker encoding sequence.

[0113] In one aspect, the present disclosure provides recombinant nucleic acids comprising:(1) a polynucleotide sequence encoding a CARD-containing protein or a functional fragment thereof, and (2) a polynucleotide sequence encoding the recombinant polypeptide of the disclosure, wherein the recombinant nucleic acid is an RNA construct.

[0114] In one aspect, the present disclosure provides recombinant nucleic acids comprising: (1) a polynucleotide sequence encoding a CARD-containing protein or a functional fragment thereof, (2) a polynucleotide sequence encoding the recombinant polypeptide of the disclosure, and (3) a polynucleotide encoding an additional potency enhancement polypeptide, a cytokine, a chemokine, or a growth factor, wherein the nucleic acid is an RNA construct.

[0115] In one aspect, the present disclosure provides combinations of recombinant nucleic acid constructs comprising: (1) a first nucleic acid construct encoding a CARD-containing protein or a functional fragment thereof, and (2) a second nucleic acid construct encoding the recombinant polypeptide of the disclosure, wherein the first and the second nucleic acid constructs are both RNA constructs.

[0116] In one aspect, the present disclosure provides combinations of recombinant nucleic acid constructs comprising: (1) a first nucleic acid construct encoding a CARD-containing protein or a functional fragment thereof, (2) a second nucleic acid construct encoding the recombinant polypeptide of the disclosure, and (3) a third nucleic acid construct encoding a polynucleotide encoding an additional potency enhancement polypeptide, a cytokine, a chemokine, or a growth factor, and wherein the first, second, and third nucleic acid constructs are each RNA constructs.

[0117] In some embodiments, the CARD-containing protein comprises (i) a CARD domain comprising a sequence having at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to amino acids 11-103 of SEQ ID NO: 1 and (ii) a Src Homology region 2 (SH2) domain comprising a sequence having at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to amino acids 622-716 of SEQ ID NO: 1. In some embodiments, the CARD containing protein comprises a sequence having at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to any one of SEQ ID NO: 1-4. In some embodiments, the second recombinantpolypeptide comprising the CARD containing protein or the functional fragment thereof comprises no more than 500, no more than 550, no more than 600, no more than 650, or no more than 700 amino acids.

[0118] In one aspect, the present disclosure provides a vector comprising the recombinant nucleic acid of the disclosure. In some embodiments, the vector is a plasmid. In some embodiments, the vector is a single stranded DNA vector or a double stranded DNA vector. In some embodiments, the vector is a synthetic DNA vector. In some embodiments, the vector is a linear DNA vector. In some embodiments, the vector is a closed linear DNA vector. In some embodiments, the vector is a synthetic RNA vector. In some embodiments, the vector is a phagemid vector.

[0119] In some embodiments, the vector is a viral vector. In some embodiments, the viral vector is selected from a retrovirus vector, an adenovirus vector, and an adeno-associated virus (AAV) vector; optionally, wherein the viral vector is the AAV vector. In some embodiments, the retrovirus is a lentivirus. In some embodiments, the lentivirus is a VSV-G pseudotyped lentivirus. In some embodiments, the AAV vector is AAV6 vector. In some embodiments, the AAV vector is AAV9 vector. In some embodiments, the AAV vector is a split-intein dual AAV vector. In some embodiments, the vector is a redirected lentiviral vector.

[0120] In some embodiments, the vector is a fusosome.

[0121] In some embodiments, the vector is a lentiviral particle engineered with the anti-CD3 Cocal glycoprotein.

[0122] In some embodiments, the vector is an enveloped delivery vehicle.

[0123] In some embodiments, the vector is self-replicating RNA virus.

[0124] In some embodiments, the vector is mRNA-packaging virus-like particle.

[0125] In some embodiments, the vector is RNP-packaging virus-like particle.

[0126] In one aspect, the present disclosure provides particles comprising the recombinant nucleic acid of the disclosure.

[0127] In some embodiments, the particle is a lipid nanoparticle (LNP). In some embodiments, the particle is a selective organ targeting (SORT) LNP. In some embodiments, the particle is an antibody targeted LNP. In some embodiments, the LNP comprises: (i) an ionizable lipid (e.g., an amino lipid), (ii) a sterol or other structural lipid, (iii) a non-cationic helper lipid or phospholipid, and (iv) a PEG-lipid (e.g., a PEG-modified lipid).

[0128] In some embodiments, the particle is a polymer nanoparticle.

[0129] In some embodiments, the particle is a protein nanoparticle.

[0130] In one aspect, the present disclosure provides engineered cells, wherein the engineered cell (i) comprises the recombinant nucleic acid of the disclosure (ii) is transfected or transduced by the vector of the disclosure, or (iii) is contacted by the particle of the disclosure.

[0131] In one aspect, the present disclosure provides engineered cells expressing the modified peptide epitope of the disclosure, or the recombinant polypeptide of the disclosure.

[0132] In one aspect, the present disclosure provides engineered cells expressing (i) a recombinant receptor that comprises a binding region for an antigen, and (ii) a recombinant polypeptide comprising a tag in its extracellular region, wherein the tag comprises a peptide epitope or a mimotope that mimics an epitope.

[0133] In one aspect, the present disclosure provides engineered cell comprising the recombinant polypeptide of the disclosure or the recombiant nucleic acid of the disclosure.

[0134] In some embodiments, the engineered cell further expresses a second recombinant polypeptide comprising a caspase-associated recruitment domain (CARD) containing protein or a functional fragment thereof. In some embodiments, the second recombinant polypeptide comprising the CARD containing protein or the functional fragment thereof comprises no more than 500, no more than 550, no more than 600, no more than 650, or no more than 700 amino acids.

[0135] In some embodiments, the engineered cell comprises one or more additional peptide epitope(s) or mimotope(s) present on the cell surface, wherein the one or more additional peptide epitope(s) or mimotope(s) are the same as the peptide epitope or mimotope or different from the peptide epitope or mimotope.

[0136] In some embodiments, activating the engineered cell results in the secretion of IL-2 by the cell at a level that is at least 10%, at least 20%, at least 50%, at least 1-fold, at least 2- fold, at least 5-fold, at least 10-fold, at least 20-fold, least 30-fold, or at least 100-fold higher than the IL-2 secretion level of a control cell lacking the recombinant polypeptide.

[0137] In some embodiments, expression of the recombinant polypeptide is under the control of an endogenous promoter. In some embodiments, the endogenous promoter is a TCRa promoter, a TCRb promoter, a CD3d promoter, a CD3g promoter, a CD3e promoter, a CD3z promoter, a CARD9 promoter, a CARDIO promoter, a CARD 11 promoter, a CARD 14 promoter, or a PIK3R3 promoter.

[0138] In some embodiments, expression of the recombinant polypeptide is under the control of an exogenous promoter. In some embodiments, the exogenous promoter is or comprises a minimal TATA promoter, pGK promoter, actin promoter, CD4 promoter, CD8a promoter, CD8b promoter, TCRa promoter, TCRb promoter, CD3d promoter, CD3g promoter, CD3e promoter, CD3z promoter, CARD9 promoter, CARDIO promoter, CARD 11 promoter, CARD 14 promoter, PIK3R3 promoter, CD25 promoter, IL2 promoter, IL7 promoter, IL 15 promoter, KLRG-1 promoter, HLA-DR promoter, CD38 promoter, CD69 promoter, Ki-67 promoter, CDl la promoter, CD58 promoter, CD99 promoter, CD62L promoter, CD 103 promoter, CCR4 promoter, CCR5 promoter, CCR6 promoter, CCR9 promoter, CCR10 promoter, CXCR3 promoter, CXCR4 promoter, CL A promoter, Granzyme A promoter, Granzyme B promoter, Perforin promoter, CD57 promoter, CD161 promoter, IL-18Ra promoter, CD69 promoter, GzmB promoter, T-bet promoter, IFNgamma promoter, TIM3 promoter, IL4 promoter, GAT A3 promoter, IL1 promoter, IL5 promoter, IL6 promoter, IL 13 promoter, IL 10 promoter, IL17A promoter, IL6 promoter, IL21 promoter, IL23R promoter, FoxP3 promoter, CTLA4 promoter, CD25 promoter, PD1 promoter, CD45RO promoter, CCR7 promoter, CD28 promoter, CD95 promoter, CD28 promoter, CD27 promoter, CD 127 promoter, PD-1 promoter, CD 122 promoter, CD 132 promoter, c-Kit promoter, nuclear factor of activated T cells (NF AT) promoter, programmed death 1 (PD-1) promoter, T cell immunoglobulin mucin-3 (TIM-3) promoter, cytotoxic T lymphocyte antigen-4 (CTLA4) promoter, lymphocyte-activation protein 3 (LAG-3) promoter, tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) promoter, B- and T-lymphocyte attenuator (BTLA) promoter, CD25 promoter, CD69 promoter, Fas ligand (FasL) promoter, TIGIT promoter, TGF-beta promoter, T-bet promoter, Eomes promoter, GATA3 promoter, CD45RA promoter, 2B4 promoter, Type I interferon (IFN) alpha, Type I IFN beta promoter, IFN gamma promoter, IRF3 promoter, IRF7 promoter, NFkB promoter, AP-1 promoter, TNF-alpha promoter, CD 130 promoter, NR4A1 promoter, NR4A2, NR4A3 promoter, MND promoter, EF-1 alpha promoter, short EF-1 alpha promoter, CAG promoter, ubiquitin / S27a promoter, SV40 promoter, SV40 early promoter, adenovirus major late promoter, mouse metallothionein- 1 promoter, Moloney murine leukemia virus (MMLV) long terminal repeat (LTR) region, CMV promoter, immunoglobulin promoter, heat shock promoter, polyoma virus promoter, fowlpox virus promoter, bovine papilloma virus promoter, avian sarcoma virus promoter, retrovirus promoter, hepatitis-B virus promoter, PGK promoter, vaccinia virus 7.5K promoter, TK promoter of HSV, mousemammary tumor virus (MMTV) promoter, LTR promoter of HIV, promoter of 29oloney virus, Epstein Barr virus (EBV) promoter, Rous sarcoma virus (RSV) promoter, U6 promoter, or UBC promoter.

[0139] In some embodiments, the nucleic acid construct is inserted in a T-cell receptor (TCR) locus, a CD3 locus, a B2 microglobulin (B2M) locus, a class II transactivator (CIITA) locus, a CARD9 locus, a CARDIO locus, a CARD11 locus, a CARD14 locus, a PIK3R3 locus, or a safe harbor locus. In some embodiments, the TCR locus is or comprises a TRAC locus, a TRBC1 locus, a TRBC2 locus, or a TRAJ locus. In some embodiments, the TRAJ locus is or comprises a TRAJ intron splice acceptor locus.

[0140] In some embodiments, the cell does not express a functional gene product of an endogenous TRAC locus. In some embodiments, the cell does not express a functional gene product of an endogenous TRBC1 locus. In some embodiments, the cell does not express a functional gene product of an endogenous TRBC2 locus. In some embodiments, the cell does not express a functional gene product of an endogenous TRAJ locus. In some embodiments, the CD3 locus is or comprises: a CD3d locus, a CD3g locus, a CD3e locus, or CD3z locus. In some embodiments, the cell does not express a functional gene product of an endogenous CD3d locus. In some embodiments, the cell does not express a functional gene product of an endogenous CD3g locus. In some embodiments, the cell does not express a functional gene product of an endogenous CD3e locus. In some embodiments, the cell does not express a functional gene product of an endogenous CD3z locus. In some embodiments, the safe harbor locus is or comprises an AAVS1, ABO, CCR5, CLYBL, CXCR4, F3, FUT1, HMGB1, KDM5D, LRP1, MICA, MICB, RHD, ROSA26, or SHS231 locus.

[0141] In some embodiments, the nucleic acid construct is inserted is an exon, an intron, between an intron and an exon, or a regulatory region.

[0142] In some embodiments, the cell is selected from the group of consisting of a T cell, a CD4+ T cell, a CD8+ T cell, a regulatory T cell (Treg), a gamma delta T cell (yST), an invariant natural killer T (iNKT) cell, a mucosal associated invariant T (MAIT) cell, a macrophage, a monocyte, a natural killer (NK) cell, a tumor infiltrating lymphocyte (TIL), a cytotoxic T cell, a T helper cell, a memory T cell, a central memory T (TCM) cell, a stem memory T (TSCM) cell, a stem-cell-like memory T cell (or stem-like memory T cells), an effector memory T (TEM) cell, a TEMRA (CD45RA+) cell, an effector T cell, a Thl cell, a Th2 cell, a Th9 cell, a Thl7 cell, a Th22 cell, a Tfh (follicular helper) cell, a natural killer T(NKT) cell, a transitional memory T (TTM) cell, a terminal effector T (TTE) cell, a naive T (TN) cell, a hematopoietic stem cell, and a progenitor cell of the lymphoid lineage.

[0143] In some embodiments, the cell is selected from the group consisting of a T cell, a macrophage, a monocyte, and a natural killer (NK) cell. In some embodiments, the cell is a T cell. In some embodiments, the T cell is selected from the group consisting of a regulatory T cell (Treg), a gamma delta T cell, a CD8+ T cell, an invariant iNKT cell, a MAIT cell, a CAR T cell, a tumor-infiltrating lymphocyte, and an engineered T cell comprising a transcriptional receptor.

[0144] In some embodiments, the cell is an autologous cell.

[0145] In some embodiments, the cell is an allogeneic cell.

[0146] In some embodiments, the cell is a primary cell.

[0147] In some embodiments, the cell is derived from a stem cell.

[0148] In some embodiments, the cell is genetically modified.

[0149] In some embodiments, the cell has reduced or eliminated cell surface expression of an endogenous T cell receptor.

[0150] In some embodiments, the cell is more prone to display central memory phenotype than a control cell lacking the recombinant polypeptide.

[0151] In some embodiments, the cell further comprises one or more modifications that inactivate or disrupt one or more alleles of TGFbeta, Regnase-1, FAS, PTPN2, NR4A3, CD52, PD-1, B2M, CIITA, SOCS1, CBLB, DGKalpha, and / or DGK^.

[0152] In some embodiments, the cell is a mammalian cell.

[0153] In some embodiments, the cell is selected from the group consisting of a human cell, a mouse cell, and a canine cell.

[0154] In some embodiments, the cell further comprises a sequence specific nuclease, a nucleic acid programmable DNA binding protein, an RNA guided nuclease, an RNA-guided nuclease comprising a Cas nuclease and a guide RNA (CRISPR-Cas combination), a ribonucleoprotein (RNP) complex comprising a gRNA and a Cas nuclease, a homing endonuclease, a zinc finger nuclease (ZF) nucleic acid binding entity, a transcription activator-like effector (TALE) nucleic acid binding entity, a meganuclease, a Cas nuclease, a core Cas protein, a homing endonuclease, an endonuclease-deficient-Cas protein, an enzymatically inactive Cas protein, a CRISPR- associated transposase (CAST), a Type II or Type V Cas protein, or a functional portion thereof. In some embodiments, the RNA-guidednuclease is a Life Edit nuclease (LEG). In some embodiments, the RNA-guided nuclease is a LEG14 nuclease.

[0155] In some embodiments, the cell further comprises Casl, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8a, Cas8b, Cas8c, Cas9, CaslO, Casl2, Casl2a (Cpfl), Casl2b (C2cl), Casl2c (C2c3), Casl 2d (CasY), Casl2e (CasX), Casl2f (C2cl0), Cas 12g, Casl2h, Casl2i, Cas 12k (C2c5), Casl3, Casl3a (C2c2), Casl3b, Casl3c, Casl3d, C2c4, C2c8, C2c9, Cmrl, Cmr2, Cmr3, Cmr4, Cmr5, Cmr6, Csdl, Csd2, Cas5d, Csel, Cse2, Cse3, Cse4, Cas5e, Csfl, Csml, Csm2, Csm3, Csm4, Csm5, Csnl, Csn2, Cstl, Cst2, Cas5t, Cshl, Csh2, Cas5h, Csal, Csa2, Csa3, Csa4, Csa5, Cas5a, CsxlO, Csxl 1, Csyl, Csy2, Csy3, Csy4, Mad7, SpCas9, eSpCas9, SpCas9-HFl, HypaSpCas9, HeFSpCas9, evoSpCas9 high-fidelity variants of SpCas9, SaCas9, NmeCas9, CjCas9, StCas9, TdCas9, LbCasl2a, AsCasl2a, AacCasl2b, BhCasl2b v4, TnpB, dCas (D10A), dCas (H840A), dCasl3a, dCasl3b, or a functional fragment thereof.

[0156] In some embodiments, the engineered cell exhibits one or more of the following: (a) improved binding to the antigen expressed on the surface of a target cell; (b) reduced recombinant polypeptide-induced antigen-independent signaling and / or activation of the engineered cell; and / or (c) reduced cross-linking between the recombinant polypeptides expressed on the cell surface of the engineered cell; compared to a control cell expressing a recombinant polypeptide comprising the unmodified peptide epitope or mimotope. In some embodiments, the engineered cell exhibits one or more of the following: (a) comparable levels of binding to the antigen expressed on the surface of a target cell; (b) comparable levels of recombinant polypeptide-induced antigen-independent signaling and / or activation of the engineered cell; and / or (c) comparable levels of cross-linking between the recombinant polypeptides expressed on the cell surface of the engineered cell; compared to a control cell expressing a recombinant polypeptide that does not comprise the peptide epitope or mimotope.

[0157] In some embodiments, the cell has reduced exhaustion, increased proliferative capacity, enhanced replicative lifespan, decreased replicative senescence, enhanced antitumor effect, reduced dysfunction, enhanced persistence, and / or increase intratumoral presence in vivo.

[0158] In some embodiments, the cell has increased or decreased signaling through the CARD1 1-BCL10-MALT1 complex, NF-KB, AP-1, NF AT, JAK / STAT, and / or MEK / ERK pathways.

[0159] In one aspect, the present disclosure provides compositions comprising the vector of the disclosure, the particle of the disclosure, or the engineered cell of the disclosure. In some embodiments, the composition further comprises a pharmaceutically acceptable carrier.

[0160] In some embodiments, at least at or about 70%, 75%, 80%, 85%, 90%, or 95% of the engineered cells, or of the total cells, in the composition express the recombinant polypeptide.

[0161] In some embodiments, at least at or about 70%, 75%, 80%, 85%, 90%, or 95% of the engineered cells, or of the total cells, in the composition do not express a gene product of an endogenous TRAC locus.

[0162] In some embodiments, at least at or about 70%, 75%, 80%, 85%, 90%, or 95% of the engineered cells, or of the total cells, in the composition express the recombinant polypeptide of the disclosure; at least at or about 70%, 75%, 80%, 85%, 90%, or 95% of the engineered cells, or of the total cells, in the composition express the receptor, or recombinant TCR; and / or at least at or about 70%, 75%, 80%, 85%, 90%, or 95% of the engineered cells, or of the total cells, in the composition do not express a gene product of an endogenous TRAC locus.

[0163] In some embodiments, at least at or about 70%, 75%, 80%, 85%, 90%, or 95% of the engineered cells, or of the total cells, in the composition do not express a functional gene product of an endogenous TRAJ locus. In some embodiments, at least at or about 70%, 75%, 80%, 85%, 90%, or 95% of the engineered cells, or of the total cells, in the composition express the recombinant polypeptide of the disclosure. In some embodiments, at least at or about 70%, 75%, 80%, 85%, 90%, or 95% of the engineered cells, or of the total cells, in the composition express the receptor (e.g., the CAR, T-body, HIT receptor, or TCR). In some embodiments, at least at or about 70%, 75%, 80%, 85%, 90%, or 95% of the engineered cells, or of the total cells, in the composition do not express a functional gene product of an endogenous TRAJ locus.

[0164] In some embodiments, the composition comprises CD4+ T cells and CD8+ T cells, and the percentage of CD4+ T cells in the composition is between at or about 20% and at or about 80%, or at or about 20%, 30%, 40%, 50%, 60%, 70%, or 80% of the total cells in the composition; and / or the percentage of CD8+ T cells in the composition is between at or about 20% and at or about 80%, or at or about 20%, 30%, 40%, 50%, 60%, 70%, or 80% ofthe total cells in the composition; and / or the ratio of CD4+ T cells to CD8+ T cells is from at or about 1 :3 to at or about 3 : 1. In some embodiments, the ratio is at or about 1 : 1.

[0165] In one aspect, the present disclosure provides methods of preparing an engineered cell, wherein the method comprises contacting the cell with the recombinant nucleic acid of the disclosure, the vector of the disclosure or the particle of the disclosure.

[0166] In one aspect, the present disclosure provides methods of preparing an engineered cell, wherein the method comprises expressing in the cell: (a) the recombinant polypeptide of the disclosure; or (b) at least the following two polypeptides: (i) a recombinant receptor that comprises a binding region for an antigen, and (ii) a recombinant polypeptide comprising at least one tag in its extracellular region, wherein the at least one tag comprises a peptide epitope or a mimotope that mimics an epitope.

[0167] In some embodiments, the method comprises enriching the engineered cells expressing the recombinant polypeptide using a binding agent that binds to the peptide epitope or mimotope. In some embodiments, the binding agent comprises a monoclonal antibody.

[0168] In some embodiments, at least at or about 70%, 75%, 80%, 85%, 90%, or 95% of the engineered cells, or of the total cells, in the composition express the recombinant polypeptide.

[0169] In some embodiments, the cell is in vivo. In some embodiments, the cell is prepared in vivo. In some embodiments, the cell is prepared in the body of a subject in need of treatment.

[0170] In some embodiments, the recombinant polypeptide is expressed in the cell, and / or the nucleic acid construct is introduced into the cell, by contacting the cell with a vector or a particle.

[0171] In some embodiments, the vector is a plasmid, a synthetic DNA vector, a single stranded DNA vector, a double stranded DNA vector, a linear DNA vector, a closed linear DNA vector, a synthetic RNA vector, a phagemid vector, an RNA vector, an mRNA vector, or a viral vector. In some embodiments, the viral vector is selected from a retrovirus vector, an adenovirus vector, and an adeno-associated virus (AAV) vector; optionally, wherein the viral vector is the AAV vector. In some embodiments, the retrovirus is a lentivirus. In some embodiments, the lentivirus is a VSV-G pseudotyped lentivirus. In some embodiments, the AAV vector is an AAV6 vector or an AAV9 vector.

[0172] In some embodiments, the vector is a fusosome, a lentiviral particle engineered with the anti-CD3 Cocal glycoprotein, or an enveloped delivery vehicle.

[0173] In some embodiments, the vector is a self-replicating RNA virus, an mRNA- packaging virus-like particle, or a ribonucleoprotein (RNP)-packaging virus-like particle.

[0174] In some embodiments, the particle is a lipid nanoparticle (LNP), a selective organ targeting (SORT) LNP, or an antibody targeted LNP. In some embodiments, the LNP comprises: (i) an ionizable lipid (e.g., an amino lipid), (ii) a sterol or other structural lipid, (iii) a non-cationic helper lipid or phospholipid, and (iv) a PEG-lipid e.g., a PEG-modified lipid).

[0175] In some embodiments, the particle is a polymer nanoparticle or a protein nanoparticle.

[0176] In some embodiments, the cell is ex vivo.

[0177] In some embodiments, the cell is prepared ex vivo.

[0178] In some embodiments, the cell is prepared outside of the body of a subject in need of treatment.

[0179] In some embodiments, the nucleic acid construct is introduced into the cell by electroporation or transfection. In some embodiments, the transfection is lipofection.

[0180] In some embodiments, the nucleic acid construct is a plasmid, a synthetic DNA vector, a single stranded DNA vector, a double stranded DNA vector, a linear DNA vector, a closed linear DNA vector, a synthetic RNA vector, a phagemid vector, an RNA vector, or an mRNA vector.

[0181] In some embodiments, the nucleic acid construct is introduced via a viral vector. In some embodiments, the viral vector is selected from a retrovirus vector, an adenovirus vector, and an adeno-associated virus (AAV) vector; optionally, wherein the viral vector is the AAV vector. In some embodiments, the retrovirus is a lentivirus. In some embodiments, the lentivirus is a VSV-G pseudotyped lentivirus. In some embodiments, the AAV vector is an AAV6 vector or an AAV9 vector.

[0182] In one aspect, the present disclosure provides methods of preparing an engineered cell, wherein the method comprises contacting the cell with an AAV vector comprising a recombinant nucleic acid encoding:(a) the recombinant polypeptide of the disclosure; or(b) at least the following three polypeptides: (i) a recombinant receptor that comprises a binding region for an antigen, (ii) a recombinant polypeptide comprising at leastone tag in its extracellular region, wherein the at least one tag comprises a peptide epitope or a mimotope that mimics an epitope; and (iii) a potency enhancement polypeptide.

[0183] In one aspect, the present disclosure provides methods of preparing an engineered cell, wherein the method comprises contacting the cell with a particle comprising a recombinant nucleic acid encoding:(a) the recombinant polypeptide of the disclosure; or(b) at least the following three polypeptides: (i) a recombinant receptor that comprises a binding region for an antigen, (ii) a recombinant polypeptide comprising at least one tag in its extracellular region, wherein the at least one tag comprises a peptide epitope or a mimotope that mimics an epitope; and (iii) a potency enhancement polypeptide. In some embodiments, the particle is a lipid nanoparticle (LNP).

[0184] In some embodiments, the method comprises genetically modifying the cell for expression of the recombinant polypeptide.

[0185] In some embodiments, the method comprises introducing into the cell one or more nucleic acids encoding a sequence specific nuclease or a nucleic acid programmable DNA binding protein. In some embodiments, the method additionally comprises introducing into the cell one or more guide RNAs. In some embodiments, the sequence-specific nuclease is an RNA guided nuclease. In some embodiments, the sequence-specific nuclease or nucleic acid programmable DNA binding domain is a homing endonuclease, a zinc finger nuclease (ZF) nucleic acid binding entity, a transcription activator-like effector (TALE) nucleic acid binding entity, a meganuclease, a Cas nuclease, a core Cas protein, a homing endonuclease, an endonuclease-deficient-Cas protein, an enzymatically inactive Cas protein, a CRISPR- associated transposase (CAST), a Type II or Type V Cas protein, or a functional portion thereof. In some embodiments, the Cas nuclease is Casl, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8a, Cas8b, Cas8c, Cas9, CaslO, Casl2, Casl2a (Cpfl), Casl2b (C2cl), Casl2c (C2c3), Casl 2d (CasY), Casl2e (CasX), Casl2f (C2cl0), Cas 12g, Casl2h, Casl2i, Cas 12k (C2c5), Casl3, Casl3a (C2c2), Casl3b, Casl3c, Casl3d, C2c4, C2c8, C2c9, Cmrl, Cmr2, Cmr3, Cmr4, Cmr5, Cmr6, Csdl, Csd2, Cas5d, Csel, Cse2, Cse3, Cse4, Cas5e, Csfl, Csml, Csm2, Csm3, Csm4, Csm5, Csnl, Csn2, Cstl, Cst2, Cas5t, Cshl, Csh2, Cas5h, Csal, Csa2, Csa3, Csa4, Csa5, Cas5a, CsxlO, Csxl 1, Csyl, Csy2, Csy3, Csy4, Mad7, SpCas9, eSpCas9, SpCas9-HFl, HypaSpCas9, HeFSpCas9, and evoSpCas9 high-fidelity variants of SpCas9, SaCas9, NmeCas9, CjCas9, StCas9, TdCas9, LbCasl2a, AsCasl2a, AacCasl2b, BhCasl2b v4, TnpB, dCas (D10A), dCas (H840A), dCasl3a, dCasl3b, or a functional portion thereof.In some embodiments, the RNA-guided nuclease is a Life Edit nuclease (LEG). In some embodiments, the LEG is a LEG14 nuclease.

[0186] In some embodiments, the method comprises introducing into the cell a ribonucleoprotein (RNP) complex comprising the sequence-specific nuclease or nucleic acid programmable DNA binding.

[0187] In some embodiments, introducing the nucleic acid construct comprises contacting the cell with the particle of the disclosure or the vector of the disclosure.

[0188] In some embodiments, the nucleic acid construct is introduced via electroporation. In some embodiments, the nucleic acid construct is introduced via AAV. In some embodiments, the nucleic acid construct is introduced via lentivirus. In some embodiments, the lentivirus is a VSV-G pseudotyped lentivirus. In some embodiments, the nucleic acid construct is introduced via LNP.

[0189] In some embodiments, the nucleic acid construct is inserted into an endogenous locus of the cell. In some embodiments, the endogenous locus is or comprises a TRAC gene locus, a TRBC1 gene locus, a TRBC2 gene locus, a TRAJ locus, a CARD9 locus, a CARDIO locus, a CARD 11 locus, a CARD 14 locus, or a PIK3R3 locus. In some embodiments, the TRAJ locus is or comprises a TRAJ intron splice acceptor locus. In some embodiments, the endogenous locus is or comprises a TRAC gene locus. In some embodiments, the endogenous locus is or comprises a TRAJ locus. In some embodiments, the TRAJ locus is or comprises a TRAJ intron splice acceptor locus. In some embodiments, the endogenous locus is or comprises a safe harbor locus. In some embodiments, the safe harbor locus is an AAVS1, ABO, CCR5, CLYBL, CXCR4, F3, FUT1, HMGB1, KDM5D, LRP1, MICA, MICB, RHD, ROSA26, or SHS231 locus.

[0190] In some embodiments, the first insertion site is an exon. In some embodiments, the first insertion site is an intron. In some embodiments, the first insertion site is between an intron and an exon. In some embodiments, the first insertion site is in a regulatory region.

[0191] In some embodiments, the first insertion site is 25 nucleotides or less from a protospacer adjacent motif (PAM) sequence, wherein the PAM sequence is ngg, nag, ngrrt, ngrm, nnnngatt, nnnnryac, nnagaaw, naaaac, tttv, ttn, attn, tttn, gttn, or yttn and wherein:(i) r = a or g,(ii) y = c or t,(iii) w = a or t,(iv) v = a or c or g, and(v) n = a, c, t, or g.

[0192] In some embodiments, the cell comprises a modified TRAC gene locus.

[0193] In some embodiments, the cell has reduced or eliminated surface expression of an endogenous TCR.

[0194] In some embodiments, the method further comprises introducing a gene editing machinery into the cell. In some embodiments, the step of introducing comprises homology- directed repair (HDR)-mediated insertion using the gene editing machinery. In some embodiments, the gene editing machinery is CRISPR / Cas9. In some embodiments, the gene editing machinery is introduced via electroporation.

[0195] In some embodiments, the nucleic acid construct is introduced via AAV.

[0196] In some embodiments, the nucleic acid construct is introduced via LNP.

[0197] In some embodiments, the method produces an engineered cell comprising: (i) the recombinant polypeptide; (ii) the immune receptor (c.g, CAR); and (iii) the modified TRAC gene locus. In some embodiments, the method thereby produces an engineered cell comprising: (i) the recombinant polypeptide; (ii) the immune receptor (e.g., CAR); and (iii) the modified TRAJ locus.

[0198] In some embodiments, the cell is for use in a cell therapy.

[0199] In one aspect, the present disclosure provides methods of treating cancer or an autoimmune disease in a subject in need thereof, comprising administering an effective amount of the engineered cells of the disclosure to the subject. In some embodiments, the peptide epitope or mimotope is bound by an antibody for an antigen associated with the cancer or the autoimmune disease.

[0200] In some embodiments, the method comprises, before the administration, generating engineered cells according to the method of the disclosure.

[0201] In some embodiments, the engineered cells are derived from immune cells: (i) obtained and / or isolated from the subject, (ii) obtained and / or isolated from one or more donors, and / or (iii) derived from one or more stem cells.

[0202] In one aspect, the present disclosure provides methods of treating an autoimmune disease in a subject in need thereof, comprising administering an effective amount of the recombinant nucleic acid of the disclosure, the vector of the disclosure, or the particle of the disclosure, to the subject. In some embodiments, the peptide epitope or mimotope is bound by an antibody for an antigen associated with the autoimmune disease. In someembodiments, the autoimmune disease is lupus, systemic lupus erythematosus, rheumatoid arthritis, psoriasis, psoriatic arthritis, multiple sclerosis, Crohn’s disease, ulcerative colitis, Addison’s disease, Graves’ disease, Sjogren’s syndrome, Hashimoto’s thyroiditis, or celiac disease.

[0203] In one aspect, the present disclosure provides methods of treating cancer in a subject in need thereof, comprising administering an effective amount of the recombinant nucleic acid of the disclosure, the vector of the disclosure, or the particle of the disclosure, to the subject. In some embodiments, the peptide epitope or mimotope is capable of being bound by an antibody for an antigen associated with the cancer.

[0204] In some embodiments, the cancer is a hematological cancer. In some embodiments, the cancer is myeloid neoplasm, myelodysplastic syndromes (MDS), myeloproliferative / myelodysplastic syndromes, acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), chronic myelogenous leukemia (CML), blast crisis chronic myelogenous leukemia (bcCML), B-cell acute lymphoid leukemia (B-ALL), T-cell acute lymphoid leukemia (T-ALL), T-cell lymphoma, or B-cell lymphoma.

[0205] In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is small cell lung cancer, colorectal cancer, testicular cancer, ovarian cancer, melanoma, lymphoma, leukemia, multiple myeloma, prostate cancer, breast cancer, nonsmall cell lung cancer, gastric cancer, esophageal cancer, liver cancer, kidney cancer, head & neck cancer, glioblastoma, neuroblastoma, soft tissue sarcoma, uterine cancer, brain cancer, skin cancer, renal cancer, bladder cancer, pancreatic cancer, thyroid cancer, eye cancer, gastrointestinal cancer, carcinoma, or sarcoma.

[0206] In some embodiments, the cancer expresses CD1, CD la, CD lb, CDlc, CD Id, CDle, CD2, CD3d, CD3e, CD3g, CD3s, CD4, CD5, CD7, CD8a, CD8b, CD19, CD20, CD21, CD22, CD23, CD24, CD25, CD27, CD28, CD30, CD33, CD34, CD38, CD40, CD44v6, CD45, CD46, CD47 CD48, CD52, CD59, CD66, CD70, CD71, CD72, CD73, CD79A, CD79B, CD80 (B7.1), CD86 (B7.2), CD94, CD95, CD97, CD123, CD134, CD140 (PDGFR4), CD152, CD154, CD158, CD171, CD178, CD179, CD179a, CD181 (CXCR1), CD182 (CXCR2), CD183 (CXCR3), CD210, CD213A2, CD246, CD252, CD253, CD261, CD262, CD272, CD273 (PD-L2), CD274 (PD-L1), CD276 (B7H3), CD279, CD295, CD339 (JAG1), CD340 (HER2), CDH17, CEA, CLECL1, CLL-1, CLDN6, CLDN18.2, CS1, DLL3, LY6G6D, GCC, p53R175H, PRAME, EGFR, EGFRvIII, FGFR2, AFP, CA125,MUC-1, MAGE, ALPI, alkaline phosphatase placental-like 2 (ALPPL2), B-cell maturation antigen (BCM A), green fluorescent protein (GFP), enhanced green fluorescent protein (eGFP), KLK2, KLK3, Mesothelin, IL13Ra2, ggcc, signal regulatory protein a (SIRPa), TCRalpha, TCRbeta, TSHR, GD2, GD3, Tn Ag, cMET, Axl, R0R1, R0R2, GPC1, GPC2, GPC3, FLT3, TAG72, CEA, EPC AM, KIT (CD117), IL-13Ra2, IL-l lRa, PSCA, PRSS21, VEGFR2, LewisY, PDGFRp, SSEA-4, folate receptor alpha, ERBB2 (Her2 / neu), MUC1, MUC16, NCAM, prostase, PAP, ELF2M, Ephrin B2, IGF-I receptor, CAIX, LMP2, gplOO, bcr-abl, tyrosinase, EphA2, STEAP1, STEAP2, fucosyl GM1, sLe, GM3, TGS5, HMWMAA, o-acetyl-GD2, folate receptor beta, TEM1 / CD248, TEM7R, R0PN1, GPRC5D, GPA33, CX0RF61, ALK, Polysialic acid, PLAC1, GloboH, NY-BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP, WT1, NY-ESO-1, LAGE-la, MAGE-A1, legumain, HPV E6,E7, MAGE Al, ETV6-AML, sperm protein 17, XAGE1, Tie 2, MAD-CT-1, MAD-CT-2, Fos-related antigen 1, p53, p53 mutant, p53R175H, KRAS, mutant KRAS, KRAS G12D, prostein, surviving, telomerase, PCTA-l / Galectin 8, MelanA / MARTl, Ras mutant, hTERT, sarcoma translocation breakpoints, ML-IAP, ERG (TMPRSS2 ETS fusion gene), NA17, PAX3, androgen receptor, cyclin Bl, MYCN, RhoC, TRP-2, CYP1B1, BORIS, SART3, PAX5, OY-TES1, LCK, AKAP-4, SSX2, RAGE-1, human telomerase reverse transcriptase, RU1, RU2, intestinal carboxyl esterase, mut hsp70- 2,LAIR1, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, FCRL5, IGLL1, PSMA, TROP2, citrullinated vimentin, the extracellular portion of the APRIL protein, or any combination thereof.

[0207] In some embodiments, the cancer expresses B7H3, BCMA, CD 19, CD20, CD22, CD70, CD79a, CD79b, CLDN6, CLDN18.2, DLL3, GCC, GD2, GD3, GPC3, GPRC5D, KLK3, LY6G6D, p53R175H, PRAME, ROPN1, STEAP1, or STEAP2. In some embodiments, the cancer expresses KLK2, Ly6G6D, CLDN6, DLL3 or CD 19.

[0208] In some embodiments, the cancer is selected from Table 2. In some embodiments, the cancer and the corresponding cancer-associated antigen are selected from Table 2.

[0209] In some embodiments, the peptide epitope or mimotope is bound by an antibody for a cancer-associated antigen of T cell lymphoma. In some embodiments, the cancer- associated antigen of T cell lymphoma is CCR4, CCR7, or CD3.

[0210] In some embodiments, the method further comprises administering an antibody to the subject, wherein the antibody binds the tag, the peptide epitope, or the mimotope.

[0211] In some embodiments, the method further comprises monitoring the subject for an adverse event.

[0212] In some embodiments, the method further comprises monitoring the subject for treatment outcomes.

[0213] In one aspect, the present disclosure provides methods of depleting engineered cells in a subject in need thereof, comprising administering to the subject an antibody that binds to a tag expressed on the surface of the engineered cells, wherein the tag comprises a peptide epitope or a mimotope that mimics an epitope.

[0214] In one aspect, the present disclosure provides methods of reducing or eliminating at least one symptom of an adverse event caused by engineered cells in a subject in need thereof, comprising administering to the subject an antibody that binds to a tag expressed on the surface of the engineered cells, wherein the tag comprises a peptide epitope or a mimotope that mimics an epitope.

[0215] In some embodiments, the subject has been administered the engineered cells within 1, 2, 3, 4, 5, or 6 days, or within 1, 2, 3, or 4 weeks, or within 1, 2, 3, 6, or 12 months, or within 1, 2, 3, 4, or 5 years, prior to administration of the antibody.

[0216] In some embodiments, the subject has been administered the recombinant nucleic acid of the disclosure, the vector of the disclosure, or the particle of the disclosure, within 1, 2, 3, 4, 5, or 6 days, or within 1, 2, 3, or 4 weeks, or within 1, 2, 3, 6, or 12 months, or within 1, 2, 3, 4, or 5 years, prior to administration of the antibody.

[0217] In some embodiments, administration of the antibody depletes at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 99%, at least 99.5%, or at least 99.9% of the engineered cells expressing the tag, the peptide epitope, or the mimotope in the subj ect.

[0218] In some embodiments, the antibody is administered after the detection or occurrence of an adverse event, and wherein administration of the antibody reduces or eliminates at least one symptom of the adverse event.

[0219] In some embodiments, the method additionally comprises administration of corticosteroids, tocilizumab, and / or anakyrin.

[0220] In some embodiments, the adverse event is cytokine release syndrome (CRS), immune effector cell-associated neurotoxicity syndrome (ICANS), hemophagocytic lymphohistiocytosis (HLH), on-target off-tumor toxicity, tumor lysis syndrome, lymphoproliferative disorder, splenomegaly, lymphadenopathy, an abnormal ratio ofengineered cells to other blood cells, acute toxicity, potency enhancement mediated cytotoxicity, potency enhancement mediated GVHD, uncontrolled expansion of engineered cells, or engineered cell mediated lymphoma.

[0221] In some embodiments, the antibody is administered after the detection or occurrence of a treatment outcome. In some embodiments, the treatment outcome is reduction or elimination of the cancer or the autoimmune disease.

[0222] In some embodiments, the method additionally comprises monitoring the subject for reduction or elimination of at least one symptom of the adverse event. In some embodiments, the monitoring comprises one or more of:Clinical laboratory tests of ferritin, C-reactive protein (CRP), creatinine, alanine transaminase (ALT), aspartate transaminase (AST), fibrinogen, lactose dehydrogenase (LDH), absolute or relative white blood cell count, and / or absolute or relative T cell count;Imaging, wherein the imaging is PET, CT, MRI, and / or x-ray;Clinical assessment of body temperature, blood pressure, blood oxygenation, heart rate, cognitive status, and / or organ function; andStandardized assessment used for detecting or monitoring one or more adverse events. In some embodiments, the standardized assessment comprises the Immune Effector Cell-Associated Encephalopathy (ICE) assessment, the Cornell Assessment, and / or the Pediatric Delirium (CAPD) assessment.

[0223] In some embodiments, the method results in at least one of:Depletion of at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% of the engineered cells in the subject;Depletion of at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% of regulatory T cells in the subject;Clinical laboratory results within the normal range for ferritin, C-reactive protein (CRP), creatinine, alanine transaminase (ALT), aspartate transaminase (AST), fibrinogen, lactose dehydrogenase (LDH), absolute or relative white blood cell count, and / or absolute or relative T cell count;Imaging results indicating a reduction in cancer or reduction in at least one symptoms of an adverse event;Reduction or absence of fever, hypotension, hypoxia, tachycardia, aphasia, cognitive impairment, coagulopathy, respiratory failure, lymphadenopathy, and / or splenomegaly;Improved score on a standardized assessment used for detecting or monitoring one or more adverse events; orAbility to reduce or eliminate administration of corticosteroids, tocilizumab, and / or anakyrin.

[0224] In some embodiments, the method results in at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% higher depletion of the engineered cell than a control method to reduce or eliminate the at least one symptom of the adverse event. In some embodiments, wherein the control method comprises administering corticosteroids, tocilizumab, and / or anakyrin.

[0225] In some embodiments, the method results in higher therapeutic efficacy than a control method to reduce or eliminate the at least one symptom of the adverse event. In some embodiments, the control method comprises administering corticosteroids, tocilizumab, and / or anakyrin.

[0226] In some embodiments, the adverse event is an acute adverse event.

[0227] In some embodiments, the antibody is a bi-specific or multi-specific antibody that is capable of specifically binding to the peptide epitope or the mimotope. In some embodiments, the antibody is a bi-specific or multi-specific antibody that is capable of binding to (i) the peptide epitope or the mimotope; and (ii) CD3. In some embodiments, the antibody is a bi-specific or multi-specific antibody that is capable of specifically binding to (i) the peptide epitope or the mimotope; and (ii) CD 16. In some embodiments, the antibody is a tri-specific or multi-specific antibody that is capable of binding to (i) the peptide epitope or the mimotope; (ii) 4 IBB or albumin, and (ii) CD3.

[0228] In some embodiments, the antibody is tarlatamab.

[0229] In some embodiments, the antibody is mogamulizumab.

[0230] In some embodiments, the antibody is CAP- 100.

[0231] In some embodiments, a dose of between about 0.003 mg / kg and about 10 mg / kg, between 0.01 mg / kg and 10 mg / kg, between 0.003 mg / kg and 0.3 mg / kg, between 0.05mg / kg and 5 mg / kg, or between 0.1 mg / kg and 1 mg / kg of the antibody is administered to the subject. In some embodiments, a dose of between about 0.003 mg / kg and about 0.999 mg / kg, about 0.003 mg / kg and about 0.01 mg / kg, about 0.01 mg / kg and about 0.03 mg / kg, about 0.03 mg / kg and about 0.1 mg / kg, about 0.1 mg / kg and about 0.3 mg / kg, about 0.3 mg / kg and about 0.6 mg / kg, or about 0.6 mg / kg and about 0.999 mg / kg, of the antibody is administered to the subject.

[0232] In some embodiments, the antibody is administered to the subject once. In some embodiments, the antibody is administered more than once. In some embodiments, the antibody is administered more than once with an interval of about 1, 2, 3, or 4 weeks.

[0233] In some embodiments, the method comprises (a) administering the engineered cell, the recombinant nucleic acid, the vector, or the particle into a brain of the subject; and (b) administering the antibody outside of the brain of the subject. In some embodiments, the method comprises intraventricular administration of the engineered cells, the recombinant nucleic acid, the vector, or the particle.

[0234] In some embodiments, the method comprises systemic administration of the antibody. In some embodiments, the method comprises intravenous administration of the antibody. In some embodiments, the method results in at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% higher depletion of the engineered cell that have crossed the blood brain barrier than a control method that reduces or eliminates the at least one symptom of the adverse event. In some embodiments, the control method comprises administering corticosteroids, tocilizumab, and / or anakyrin.

[0235] In some embodiments, the antibody is administered more than once. In some embodiments, the antibody is administered more than once with an interval of about 1, 2, 3, or 4 weeks. In some embodiments, the antibody is administered more than once with an interval of at least 1, 2, 3, or 4 weeks. In some embodiments, the antibody is administered two, three, or four times.

[0236] In some embodiments, both the antibody and the binding region of the recombinant polypeptide can bind the same antigen.

[0237] In some embodiments, the antibody is mogamulizumab, and wherein the method further comprises administering one or more of: (a) at least one retinoid; (b) interferonalpha; and / or (c) extracorporeal photopheresis (ECP). In some embodiments, wherein the retinoid and / or interferon alpha is delivered systemically.

[0238] In one aspect, the present disclosure provides methods of isolating or enriching the engineered cells of the disclosure, comprising contacting the engineered cells with a binding agent that binds to the epitope or the mimotope.

[0239] In some embodiments, the binding agent comprises an antibody.

[0240] In some embodiments, the binding agent comprises a purification marker. In some embodiments, the purification marker comprises biotin, avidin, a myc tag, an influenza hemagglutinin (HA) tag, a FLAG tag, a glutathione-S transferase (GST) tag, a V5 tag, a poly-Histidine (His) tag, a Fc tag, calmodulin binding protein (CBP), maltose-binding protein (MBP), thioredoxin, or a chitin-binding protein.

[0241] In some embodiments, the binding agent is bound to a solid substrate or a purification bead.

[0242] In some embodiments, the method results in a population of purified engineered cells in which at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% of the engineered cell expresses the recombinant polypeptide.

[0243] In one aspect, the present disclosure provides methods of treating cancer or an autoimmune disease in a subject in need thereof, comprising administering a therapeutically effective amount of engineered cells isolated or enriched by the method of the disclosure.

[0244] In one aspect, the present disclosure provides methods of ex vivo or in vivo measuring the number of the engineered cells of the disclosure, comprising administering, or contacting the engineered cells with, a binding agent that binds to the epitope or the mimotope. In some embodiments, the binding agent comprises an antibody.

[0245] In one aspect, the present disclosure provides methods of treating cancer or an autoimmune disease in a subject in need thereof, comprising: (a) administering an effective amount of an engineered cell, a recombinant nucleic acid, a vector, or a particle disclosed herein, to the subject, wherein the engineered cell or the recombinant polypeptide comprises two or more of the tags, the peptide epitopes or the mimotopes that are capable of binding to mogamulizumab, and wherein the tags, the peptide epitopes or the mimotopes are the same or different; and (b) administering at least one dose of mogamulizumab to the subject.

[0246] In some embodiments, each of the dose(s) of mogamulizumab is between about 0.003 mg / kg and about 0.999 mg / kg, about 0.003 mg / kg and about 0.01 mg / kg, about 0.01mg / kg and about 0.03 mg / kg, about 0.03 mg / kg and about 0.1 mg / kg, about 0.1 mg / kg and about 0.3 mg / kg, about 0.3 mg / kg and about 0.6 mg / kg, or about 0.6 mg / kg and about 0.999 mg / kg.

[0247] In some embodiments, the method results in at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% higher depletion of the engineered cell than a control method to reduce or eliminate the at least one symptom of the adverse event; optionally wherein the control method comprises administering corticosteroids, tocilizumab, and / or anakyrin.

[0248] In some embodiments, the engineered cell, the recombinant nucleic acid, the vector, or the particle is administered at least 1, 2, 3, 4, 5, 6, 7, 8 weeks, or at least 2, 3, 4, 5, 6, 8 or 10 months, or at least 1, 2, 3, 4, or 5 years, before the first dose of mogamulizumab is administered to the subject.

[0249] In one aspect, the present disclosure provides methods of treating cancer in a subject in need thereof, comprising: (a) intraventricularly administering an effective amount of an engineered cell, a recombinant nucleic acid, a vector, or a particle disclosed herein, to the subject; and (b) systemically administering at least one dose of mogamulizumab to the subject.

[0250] In some embodiments, each of the dose(s) of mogamulizumab is between about 0.003 mg / kg and about 0.999 mg / kg, about 0.003 mg / kg and about 0.01 mg / kg, about 0.01 mg / kg and about 0.03 mg / kg, about 0.03 mg / kg and about 0.1 mg / kg, about 0.1 mg / kg and about 0.3 mg / kg, about 0.3 mg / kg and about 0.6 mg / kg, or about 0.6 mg / kg and about 0.999 mg / kg.

[0251] In some embodiments, the method results in at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% higher depletion of the engineered cells outside the brain than a control method that reduces or eliminates the at least one symptom of the adverse event; optionally wherein the control method comprises administering corticosteroids, tocilizumab, and / or anakyrin. In some embodiments, the method comprises administering at least two, at least three, or at least four doses of mogamulizumab to the subject.BRIEF DESCRIPTION OF THE DRAWINGS

[0252] FIGS. 1A-1N Show schematics of non-limiting examples of CAR and TCR construct designs. FIG. 1A: a CAR construct without an epitope tag; FIG. IB: a CAR construct with an N-terminal epitope tag; FIG. 1C: a CAR construct with an epitope tag in the linker region; FIG. ID: a CAR construct with an epitope tag in the hinge region; FIG. IE: a TCRbeta-TCRalpha construct without an epitope tag; FIG. IF: a TCRbeta-TCRalpha construct with an N-terminal epitope tag; FIG. 1G: a TCRbeta-TCRalpha construct with an epitope tag between the TCRbeta variable and the TCRbeta constant regions; FIG. 1H: a TCRbeta-TCRalpha construct with an epitope tag in the linker region between TCRbeta and TCRalpha; FIG. II: a TCRbeta-TCRalpha construct with an epitope tag between the TCRalpha variable and the TCRalpha constant regions; FIG. 1J: a TCRalpha-TCRbeta construct without an epitope tag; FIG. IK: a TCRalpha-TCRbeta construct with an N- terminal epitope tag; FIG. IL: a TCRalpha-TCRbeta construct with an epitope tag between the TCRalpha variable and the TCRalpha constant regions; FIG. IM: a TCRalpha-TCRbeta construct with an epitope tag in the linker region between TCRalpha and TCRbeta; FIG. IN: a TCRalpha-TCRbeta construct with an epitope tag between the TCRbeta variable and the TCRbeta constant regions.

[0253] FIGS. 2A-2D show the expression of CAR constructs tagged with a bi-specific Tarlatamab antibody epitope and functionality of T cells expressing the CAR constructs against antigen (DLL3) positive cell lines. FIG. 2A is a diagram showing the percentage of CAR positive cells. FIG. 2B is a diagram showing the mean fluorescence intensity (MFI) of the cells. FIG. 2C is a diagram showing the killing of a cancer cell line that expresses DLL3 at a low level by the indicated CAR-T cells. FIG. 2D is a diagram showing the secretion of IFNgamma by the indicated CAR-T cells when co-cultured with a cancer cell line that expresses DLL3 at a low level.

[0254] FIGS. 3A-3B show the depletion and activation of a CAR-T cells expressing CAR construct that is tagged with a bi-specific tarlatamab antibody epitope. FIG. 3 A is a diagram showing the percentage of CAR positive cells. FIG. 3B is a diagram showing the percentage of CD69 positive cells in the indicated cell lines upon tarlatamab treatment.

[0255] FIGS. 4A-4F show the expression of CAR constructs with mogamulizumab epitope tags. FIGS. 4A, 4B and 4C are diagrams showing the percentage of CAR positive cells. FIGS. 4D, 4E and 4F are diagrams showing the mean fluorescence intensity (MFI) of cells expressing the CAR constructs.

[0256] FIGS. 5A-5F show the ability of different CAR-T cells expressing CAR constructs comprising a mogamulizumab epitope to be bound by mogamulizumab. FIGS. 5A, 5B and 5C are diagrams showing the percentage of T-cells that can be positively stained by mogamulizumab. FIGS. 5D, 5E and 5F are diagrams showing the mean fluorescence intensity (MFI) of the mogamulizumab staining in different CAR-T cells.

[0257] FIGS. 6A-6F show the release of cytokines of different CAR-T cells in response to antigen stimulation. FIGS. 6A, 6B and 6C are diagrams showing the production of IL-2 in various CAR-T cells. FIGS. 6D, 6E and 6F are diagrams showing the production of IFNgamma in various CAR-T cells.

[0258] FIGS. 7A-7D show the in vivo functionality of different CAR-T constructs. FIGS. 7A and 7B are diagrams showing the killing of a cancer cell line with high antigen expression by the indicated CAR-T cells. FIGS. 7C and 7D are diagrams showing the killing of a cancer cell line with low antigen expression by the indicated CAR-T cells.

[0259] FIGS. 8A-8B show the inability of a TROP2 tag to be bound by the corresponding antibody (Sacituzumab). FIG. 8A is a diagram showing the percentage of CAR positive cells. FIG. 8B is a diagram showing the percentage of cells positively stained by Sacituzumab.

[0260] FIGS. 9A-9D show that T cells expressing a CAR comprising the modified mogamulizumab epitope tag are depleted via ADCC, both in vitro and in vivo, after treatment with mogamulizumab. FIG. 9 A compares the number of GFP-positive (GFP+) T cells for untransduced T cells (UTD) compared to T cells expressing a construct that encodes both the CAR comprising the modified mogamulizumab epitope tag and GFP. FIG. 9B shows the percentage of T cells that are depleted (relative to treatment with an isotype control antibody) after 24 hour in vitro co-culture with NK cells at different T cell to NK cell ratios (2: 1, 5: 1, or 10: 1) treated with either 1) 10 ug / mL of an isotype control antibody, 2) 0.1 ug / mL of a mogamulizumab biosimilar antibody, 3) 1.0 ug / mL of a mogamulizumab biosimilar antibody, or 4) 10 ug / mL of a mogamulizumab biosimilar antibody. FIG. 9C shows cell counts in peripheral blood collected from mice treated with either: 1) untransduced T cells (UTD), 2) T cells expressing a CAR comprising the modified mogamulizumab epitope tag but not treated with a mogamulizumab biosimilar antibody (Untreated), 3) T cells expressing a CAR comprising the modified mogamulizumab epitope tag and 1 mg / kg of a mogamulizumab biosimilar antibody on day 7 after T cell infusion (Day 7 Moga Treatment), or 4) T cells expressing a CAR comprising the modifiedmogamulizumab epitope tag and 1 mg / kg of a mogamulizumab biosimilar antibody on day 14 after T cell infusion (Day 14 Moga Treatment). FIG. 9D shows cell counts in blood (Blood), bone marrow (BM), or spleen (Spleen) collected from mice 24 hours after treatment with either: 1) untransduced T cells (UTD), 2) T cells expressing a CAR comprising the modified mogamulizumab epitope tag but not treated with a mogamulizumab biosimilar antibody (Untreated), 3) T cells expressing a CAR comprising the modified mogamulizumab epitope tag and a single 1 mg / kg dose of a mogamulizumab biosimilar antibody on day 14 after T cell infusion (Single Moga Dose), or 4) T cells expressing a CAR comprising the modified mogamulizumab epitope tag and four doses of 1 mg / kg mogamulizumab biosimilar antibody, where the mogamulizumab biosimilar antibody was administered on days 5, 8, 11, and 14 after T cell infusion (Repeat Moga Dose).

[0261] FIGS. 10A-10C demonstrate that, relative to T cells expressing a CAR comprising an unmodified mogamulizumab epitope tag, T cells expressing a CAR comprising a modified mogamulizumab epitope tag, which modified epitope tag does not comprise a cystine present in the unmodified mogamulizumab epitope tag, exhibit reduced levels of antigen-independent signaling. FIGS. 10A-10B show the level of antigen-independent IFNgamma secretion (FIG. 10 A) and T cell proliferation (FIG. 10B) after T cell culture with different concentrations of a mogamulizumab biosimilar (No Moga, 1 ug / mL Moga, 10 ug / mL Moga, or 20 ug / mL Moga). The cells in this assay included T cells expressing a CAR without an epitope tag (CAR), T cells expressing a CAR comprising an unmodified mogamulizumab epitope tag (CAR.Moga), and T cells expressing a CAR comprising a modified mogamulizumab epitope tag that does not comprise a cystine present in the unmodified mogamulizumab epitope tag (CAR. Moga. del Cys). FIG. 10C shows the impact on T cell phenotype of T cells expressing a CAR comprising the unmodified mogamulizumab epitope tag or T cells expressing a CAR comprising the modified mogamulizumab epitope tag.

[0262] FIGS. 11A-11N demonstrate that a variety of CAR constructs comprising a modified mogamulizumab epitope tag are expressed by T cells at similar levels to matched untagged CAR constructs and, broadly, retain similar levels of functionality. FIGS. 11 A- 1 ID show the percentage of CAR positive T cells (FIG. 11 A) and CAR MFI (FIG. 1 IB) based on detection of the CAR and the percentage of CAR positive cells (FIG. 11C) and CAR MFI (FIG. 1 ID) based on detection of the mogamulizumab epitope tag for T cells expressing a tagged and untagged KLK2 CAR (KLK2.CAR and KLK2.moga-CAR), atagged and untagged Claudin-6 CAR (CLDN6.CAR and CLDN6.moga-CAR), a tagged and untagged DLL3 CAR (DLL3.CAR and DLL3.moga-CAR), a tagged and untagged LY6G6D CAR (LY6G6D.CAR and LY6G6D.moga-CAR), and a tagged and untagged CD 19 CAR (CD19.CAR and CD19.moga-CAR), FIGS. 1 IE-1 II show IFNgamma secretion after in vitro co-culture with target cells expressing the antigen target for each CAR for T cells expressing both tagged and untagged CARs. FIGS. 11 J-N show IL-2 secretion after in vitro co-culture with target cells expressing the antigen target for each CAR for T cells expressing both tagged and untagged CARs.

[0263] FIGS. 12A-12I show that different TCR based receptor constructs (transgenic TCRs (tTCRs) and T-bodies) comprising a modified mogamulizumab epitope tag are expressed by T cells at similar levels to matched untagged tTCRs and T-bodies and that T cells expressing tagged T-bodies retain similar levels of functionality to untagged T-bodies. FIGS. 12A-12C show the percentage of T-body positive T cells (FIG. 12A) and T-body MFI (FIG. 12B) based on detection of the T-body and the percentage of T-body positive T cells (FIG. 12C) based on detection of the mogamulizumab epitope tag. FIGS. 12D-12F show the percentage of tTCR positive T cells (FIG. 12D) and tTCR MFI (FIG. 12E) based on detection of the tTCR and the percentage of tTCR positive T cells (FIG. 12F) based on detection of the mogamulizumab epitope tag. FIGS. 12G and 12H show IFNgamma secretion (FIG. 12G) and IL-2 secretion (FIG. 12H) after in vitro co-culture with target cells expressing CD 19 for T cells expressing both tagged and untagged anti-CD19 T-body. FIG. 121 shows in vitro NK cell-mediated ADCC depletion of T cells expressing an anti-CD19 T- body comprising the modified mogamulizumab epitope tag.

[0264] In the following detailed description, reference is made to the Figures, which form a part hereof. In the Figures, similar symbols generally identify similar components, unless context dictates otherwise.

[0265] The illustrative alternatives described in the detailed description, drawings, and claims are not meant to be limiting. Other alternatives may be used and other changes may be made without departing from the spirit or scope of the subject matter presented here. It will be readily understood that the aspects and embodiments, as generally described herein, and illustrated in the Figures, can be arranged, substituted, combined, and designed in a wide variety of different configurations (for example, combined with other aspects and / or embodiments described herein), all of which are explicitly contemplated and make part of this application.DETAILED DESCRIPTION OF THE DISCLOSURE

[0266] In general, the present disclosure relates, inter alia, to compositions and methods for improving adoptive immune cell therapy. The disclosers have discovered ways to improve the control and safety of engineered immune cells by introducing to the surface of the engineered immune cells at least one depletion tag (e.g., an epitope or a mimotope) that is capable of being bound by another molecule (e.g., an antibody) to facilitate the depletion of the engineered immune cells when needed. The depletion tag may be incorporated to the extracellular region of an engineered receptor (e.g., a chimeric antigen receptor or a T cell receptor) or be presented separately on the cell surface. Upon the occurrence of an adverse event in the subject, the engineered cells can be depleted as needed by administering the corresponding tag-binding molecule (e.g., the corresponding antibody). The presence of such tag can also facilitate the sorting and / or enrichment of the engineered cells (e.g., during cell manufacturing).

[0267] At least two classes of depletion tags are disclosed herein. The first class include depletion tags that contain an epitope or mimotope derived from a biomarker of a hematologic cancer (e.g., CCR4). This strategy allows the use of therapeutic antibodies developed for hematologic cancer to effectively deplete the engineered immune cell.

[0268] The second class include depletion tags that contain an epitope or mimotope derived from the same antigen that the engineered receptor binds to (e.g., a DLL3 derived depletion tag for engineered immune cells that bind to DLL3). This strategy allows the use of the corresponding antigen-binding therapeutic antibody to simultaneously deplete the engineered immune cell and at the same time exert its anti-cancer effect.

[0269] Also disclosed are ways to improve the efficacy of the depletion tag in the engineered immune cells context, such as modifications of the epitopes or mimotopes, their placements in the engineered receptor or immune cell, and the delivery of such depletion tags or the antibodies. In some embodiments, the present disclosure relates to exploiting such depletion tags to improve the effectiveness of immune cell therapeutics. In some embodiments, the presence of the depletion tag does not negatively affect, or even improves, the efficacy of the receptor or the engineered cells that it is incorporated into. In particular, in the context of engineered receptors incorporated with the depletion tag, mutating cysteine(s) present in the epitope or mimotope may reduce the level of undesirable antigen-independent signaling and / or activation (i.e., tonic signaling) and / or improve receptor signaling capability.I. DEFINITIONS

[0270] Unless otherwise defined, all terms of art, notations, and other scientific terms or terminology used herein are intended to have the meanings commonly understood by those of skill in the art to which this application pertains. In some cases, terms with commonly understood meanings are defined herein for clarity and / or for ready reference, and the inclusion of such definitions herein should not necessarily be construed to represent a substantial difference over what is generally understood in the art. Many of the techniques and procedures described or referenced herein are well understood and commonly employed using conventional methodology by those skilled in the art. All publications, patent applications, patents, GenBank or other accession numbers and other references mentioned herein are incorporated by reference in their entirety for all purposes.

[0271] The singular form “a”, “an”, and “the” include plural references unless the context clearly dictates otherwise. For example, the term “a cell” includes one or more cells, including mixtures thereof. “A and / or B” is used herein to include all of the following alternatives: “A”, “B”, “A or B”, and “A and B .”

[0272] Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range, is encompassed within the disclosure. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges, and are also encompassed within the disclosure, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the disclosure.

[0273] Certain ranges are presented herein with numerical values being preceded by the term “approximately” or “about.” The terms “approximately” and “about” are used interchangeably and mean a quantity, level, value, number, frequency, percentage, dimension, size, amount, weight or length that varies by up to 20%, preferably up to 10%, more preferably up to 5%, and more preferably still up to 1%, of a reference quantity, level, value, number, frequency, percentage, dimension, size, amount, weight or length, inclusive of the endpoints. When the term "about" is used in conjunction with a numerical range, itmodifies that range by extending the boundaries above and below the numerical values set forth, unless otherwise apparent from context that it is impossible to extend the boundary beyond certain points (e.g., below 0% or above 100% in some cases).

[0274] As used herein, the term “comparable level” refers to a level that is in between 50% and 200%, inclusive of end points, of the reference level in the control set of conditions or circumstances, unless otherwise apparent from context that it is impossible to extend the boundary beyond certain points. In some embodiments, a comparable level may be preferably between 60% and 170%, more preferably between 70% and 140%, even more preferably between 80% and 125%, or more preferably still between 90% and 110%, of the reference level. Those of ordinary skill in the art will appreciate the appropriate control set in obtaining the reference level. In general, the appropriate control set shares identifical features with the test set except the feature(s) of interest, so that the observed results warrant a reasonable conclusion that any changes in the level observed are caused by or indicative of the variation in the feature(s) that are varied.

[0275] As will be understood by one having ordinary skill in the art, for any and all purposes, such as in terms of providing a written description, all ranges disclosed herein also encompass any and all possible sub-ranges and combinations of sub-ranges thereof. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, etc. As a nonlimiting example, each range discussed herein can be readily broken down into a lower third, middle third and upper third, etc. As will also be understood by one skilled in the art all language such as “up to,” “at least,” “greater than,” “less than,” and the like include the number recited and refer to ranges which can be subsequently broken down into sub-ranges as discussed above. Finally, as will be understood by one skilled in the art, a range includes each individual member. Thus, for example, a group having 1-3 articles refers to groups having 1, 2, or 3 articles. Similarly, a group having 1-5 articles refers to groups having 1, 2, 3, 4, or 5 articles, and so forth.

[0276] It is understood that aspects and embodiments of the disclosure described herein include “comprising,” “consisting,” and “consisting essentially of’ aspects and embodiments. As used herein, “comprising” is synonymous with “including,” “containing,” or “characterized by,” and is inclusive or open-ended and does not exclude additional, unrecited elements or method steps. As used herein, “consisting of’ excludes any elements, steps, or ingredients not specified in the claimed composition or method. As used herein,“consisting essentially of’ does not exclude materials or steps that do not materially affect the basic and novel characteristics of the claimed composition or method. Any recitation herein of the term “comprising”, particularly in a description of components of a composition or in a description of steps of a method, is understood to encompass those compositions and methods consisting essentially of and consisting of the recited components or step.

[0277] The term “percent identity,” as used herein in the context of two or more nucleic acids or proteins, refers to two or more sequences or subsequences that are the same or have a specified percentage of nucleotides or amino acids that are the same (e.g., about 60% sequence identity, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher identity over a specified region, when compared and aligned for maximum correspondence over a comparison window or designated region) as measured using a BLAST or BLAST 2.0 sequence comparison algorithms with default parameters described below, or by manual alignment and visual inspection. See e.g., the NCBI web site at ncbi.nlm.nih.gov / BLAST. Such sequences are then the to be “substantially identical.” This definition also refers to, or may be applied to, the complement of a test sequence. This definition also includes sequences that have deletions and / or additions, as well as those that have substitutions. Generally, sequence identity can exist over a region that is at least about 20 amino acids or nucleotides in length, or over a region that is 10-100 amino acids or nucleotides in length, or over the entire length of a given sequence

[0278] If necessary, sequence identity can be calculated using published techniques and widely available computer programs, such as the GCS program package (Devereux et al, Nucleic Acids Res. 12:387, 1984), BLASTP, BLASTN, FASTA (Atschul et al., J. Molecular Biol. 215:403, 1990). Sequence identity can be measured using sequence analysis software such as the Sequence Analysis Software Package of the Genetics Computer Group at the University of Wisconsin Biotechnology Center (1710 University Avenue, Madison, Wis. 53705), with the default parameters thereof.

[0279] As used herein, the term “mutation” refers to a point mutation, a gene fusion, a substitution, a gain-of-function mutation, a stop-gain mutation, an insertion mutation, a deletion mutation, a duplication mutation and / or a translocation. The mutation may be in one or more genes. The mutation may be naturally occurring. Alternatively, the mutation may be induced or engineered.

[0280] As used herein, the term “vector” refers to a recombinant nucleic acid designed for transfer between host cells, and that may be used for the purpose of transformation, e.g., the introduction of heterologous DNA into a host cell. As such, in some embodiments, the vector can be a replicon, such as a plasmid, phage, or cosmid, into which another DNA segment may be inserted so as to bring about the replication of the inserted segment. In some embodiments, the expression vector can be an integrating vector.

[0281] As used herein, the term “viral vector” refers either to a nucleic acid molecule (e.g., a transfer plasmid) that includes virus-derived nucleic acid elements that generally facilitate transfer of the nucleic acid molecule or integration into the genome of a cell or to a viral particle that mediates nucleic acid transfer. Viral particles will generally include various viral components and sometimes also host cell components in addition to nucleic acid(s). The term viral vector may refer either to a virus or viral particle capable of transferring a nucleic acid into a cell or to the transferred nucleic acid itself. Viral vectors and transfer plasmids contain structural and / or functional genetic elements that are primarily derived from a virus. Viral vectors that can be used in the disclosure include, for example, retrovirus vectors, adenovirus vectors, and adeno-associated virus vectors, lentivirus vectors, herpes virus, simian virus 40 (SV40), and bovine papilloma virus vectors (see, for example, Gluzman (Ed.), Eukaryotic Viral Vectors, CSH Laboratory Press, Cold Spring Harbor, N.Y.). For example, a recombinant polypeptide as disclosed herein can be produced in a eukaryotic host, such as a mammalian cells (e.g., COS cells, NIH 3T3 cells, or HeLa cells). These cells are available from many sources, including the American Type Culture Collection (Manassas, VA). In selecting an expression system, care should be taken to ensure that the components are compatible with one another. Artisans or ordinary skill are able to make such a determination. Furthermore, if guidance is required in selecting an expression system, skilled artisans may consult P. Jones, “Vectors: Cloning Applications”, John Wiley and Sons, New York, N.Y., 2009).

[0282] As used herein, the term “retroviral vector” refers to a viral vector or plasmid containing structural and functional genetic elements, or portions thereof, that are primarily derived from a retrovirus. The retroviral vector can be a lentiviral vector.

[0283] As used herein, the term “lentiviral vector” refers to a viral vector or plasmid containing structural and functional genetic elements, or portions thereof, including LTRs that are primarily derived from a lentivirus, which is a genus of retrovirus. Lentiviral vectors offer several attractive properties as gene-delivery vehicles, including: (i) sustained genedelivery through stable vector integration into host genome; (ii) the capability of infecting both dividing and non-dividing cells; (iii) broad tissue tropisms, including important gene- and cell-therapy -target cell types; (iv) no expression of viral proteins after vector transduction; (v) the ability to deliver complex genetic elements, such as polycistronic or intron-containing sequences; (vi) a potentially safer integration site profile; and (vii) a relatively easy system for vector manipulation and production.

[0284] As used herein, the term “pharmaceutically acceptable carrier” as used herein means any suitable carriers, diluents or excipients. These include all aqueous and nonaqueous isotonic sterile injection solutions, which may contain anti-oxidants, buffers and solutes, which render the composition isotonic with the blood of the intended recipient; aqueous and non-aqueous sterile suspensions, which may include suspending agents and thickening agents, dispersion media, antifungal and antibacterial agents, isotonic and absorption agents and the like. It will be understood that compositions of the present disclosure may also include other supplementary physiologically active agents. The carrier must be pharmaceutically “acceptable” in the sense of being compatible with the other ingredients of the composition and not injurious to the subject.

[0285] As used herein, the term “PEGylation” refers to modifying a protein by covalently attaching polyethylene glycol (PEG) to the protein, with “PEGylated” referring to a protein having a PEG attached. A range of PEG, or PEG derivative sizes with optional ranges of from about 10,000 Daltons to about 40,000 Daltons may be attached to the recombinant polypeptides of the disclosure using a variety of chemistries. In some embodiments, the average molecular weight of the PEG, or PEG derivative, is about 1 kD to about 200 kD such as, e.g., about 10 kD to about 150 kD, about 50 kD to about 100 kD, about 5 kD to about 100 kD, about 20 kD to about 80 kD, about 30 kD to about 70 kD, about 40 kD to about 60 kD, about 50 kD to about 100 kD, about 100 kD to about 200 kD, or about 150 kD to about 200 kD. In some embodiments, the average molecular weight of the PEG, or PEG derivative, is about 5 kD, about 10 kD, about 20 kD, about 30 kD, about 40 kD, about 50 kD, about 60 kD, about 70 kD, or about 80 kD. In some embodiments, the average molecular weight of the PEG, or PEG derivative, is about 40 kD.

[0286] As used herein, the terms “administration” and “administering” refer to the delivery of a bioactive composition or formulation by an administration route including, but not limited to, oral, intravenous, intra-arterial, intramuscular, intraperitoneal, subcutaneous,intramuscular, and topical administration, or combinations thereof. The term includes, but is not limited to, administering by a medical professional and self-administering.

[0287] As used herein, the term “injection” includes intravenous, intramuscular, intraarterial, intrathecal, intraventricular, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, intracerebrospinal, and intrastemal injection and infusion.

[0288] The term “cancer” generally refers to the presence of cells possessing characteristics typical of cancer-causing cells, such as uncontrolled proliferation, immortality, metastatic potential, rapid growth and proliferation rate, and certain characteristic morphological features. Cancer cells can be in the form of a tumor, but such cells can exist alone within an animal subject, or can be a non-tumorigenic cancer cell, such as a leukemia cell. These terms include a solid tumor, a soft tissue tumor, or a metastatic lesion. As used herein, the term “cancer” includes premalignant, as well as malignant cancers. In some embodiments, the cancer is a solid tumor, a soft tissue tumor, or a metastatic lesion.

[0289] As used herein, and unless otherwise specified, a “therapeutically effective” or “pharmaceutically effective” amount or number of a subject construct, nucleic acid, cell, or composition of the disclosure generally refer to an amount or number sufficient for a construct, nucleic acid, cell, or composition to accomplish a stated purpose relative to the absence of the composition, e.g., to provide a therapeutic benefit in the treatment or management of the cancer, or to delay or minimize one or more symptoms associated with the cancer. A therapeutically effective amount of a compound means an amount of therapeutic agent, alone or in combination with other therapeutic agents, which provides a therapeutic benefit in the treatment or management of the cancer. The term “therapeutically effective amount” can encompass an amount that improves overall therapy, reduces or avoids symptoms or causes of the cancer, or enhances the therapeutic efficacy of another therapeutic agent. An example of an “effective amount” is an amount sufficient to contribute to the treatment, prevention, or reduction of a symptom or symptoms of a disease, which could also be referred to as a “therapeutically effective amount.” A “reduction” of a symptom means decreasing of the severity or frequency of the symptom(s), or elimination of the symptom(s). The exact amount of a composition including a “therapeutically effective amount” will depend on the purpose of the treatment, and will be ascertainable by one skilled in the art using known techniques (see, e.g., Lieberman, Pharmaceutical DosageForms (vols. 1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999); and Remington: The Science and Practice of Pharmacy, 20th Edition, 2003, Gennaro, Ed., Lippincott, Williams & Wilkins)

[0290] As used herein, a “subject” or an “individual” includes animals, such as human (e.g., human subjects) and non-human animals. In some embodiments, a “subject” or “individual” is a patient under the care of a physician. Thus, the subject can be a human patient or an individual who has, is at risk of having, or is suspected of having a disease of interest (e.g., cancer) and / or one or more symptoms of the disease. The subject can also be an individual who is diagnosed with a risk of the condition of interest at the time of diagnosis or later. The term “non-human animals” includes all vertebrates, e.g., mammals, e.g., rodents, e.g., mice, e.g. non-human primates, and non- mammals, e.g., sheep, dogs, cows, chickens, amphibians, reptiles, etc.

[0291] The term “antibody” herein is used in the broadest sense and includes polyclonal and monoclonal antibodies, including intact antibodies and functional (antigen-binding) antibody fragments, including fragment antigen binding (Fab) fragments, F(ab’)2 fragments, Fab’ fragments, Fv fragments, recombinant IgG (rlgG) fragments, heavy chain variable (VH) regions capable of binding the antigen, single chain antibody fragments, including single chain variable fragments (scFv), and single domain antibodies ( 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, and heteroconjugate antibodies, multispecific, e.g., bispecific or trispecific, antibodies, diabodies, triabodies, and tetrabodies, tandem di-scFv, tandem tri-scFv. Unless otherwise stated, the term “antibody” should be understood to encompass functional antibody fragments thereof also referred to herein as “antigen-binding fragments.” The term also encompasses intact or full-length antibodies, including antibodies of any class or sub-class, including IgG and sub-classes thereof, IgM, IgE, IgA, and IgD.

[0292] The terms “complementarity determining region,” and “CDR,” synonymous with “hypervariable region” or “HVR,” are known in the art to refer to non-contiguous sequences of amino acids within antibody variable regions, which confer antigen e.g., specificity and / or binding affinity. In general, there are three CDRs in each heavy chain variable region (CDR- Hl, CDR- H2, CDR-H3) and three CDRs in each light chain variable region (CDR-L1, CDR-L2, CDR- L3). “Framework regions” and “FR” are known in the art to refer to thenon-CDR portions of the variable regions of the heavy and light chains. In general, there are four FRs in each full- length heavy chain variable region (FR-H1, FR-H2, FR-H3, and FR- H4), and four FRs in each full-length light chain variable region (FR-L1, FR-L2, FR-L3, and FR-L4).

[0293] The precise amino acid sequence boundaries of a given CDR or FR can be readily determined using any of a number of well-known schemes, including those described by 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 el ah, “IMGT unique numbering for immunoglobulin and T cell receptor variable domains and Ig superfamily V-like domains,” Dev Comp Immunol, 2003 Jan;27(l):55-77 (“IMGT” numbering scheme); Honegger A and Pliickthun 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 el ah, “Modeling antibody hypervariable loops: a combined algorithm,” PNAS, 1989, 86(23):9268-9272, (“AbM” numbering scheme).

[0294] 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 alignments, while the Chothia scheme is based on structural information. Numbering for both the Kabat and Chothia schemes is based upon the most common antibody region sequence lengths, with insertions accommodated by insertion letters, for example, “30a,” and deletions appearing in some antibodies. The two schemes place certain insertions and deletions (“indels”) at different positions, resulting in differential numbering. The Contact scheme is based on analysis of complex crystal structures and is similar in many respects to the Chothia numbering scheme.

[0295] The AbM scheme is a compromise between Kabat and Chothia definitions based on that used by Oxford Molecular’s AbM antibody modeling software.

[0296] Table 1, below, lists exemplary position boundaries of CDR-L1, CDR-L2, CDR- L3 and CDR-H1, CDR-H2, CDR-H3 as identified by 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 CDRs, for example, with FR-L1located before CDR-L1, FR-L2 located between CDR-L1 and CDR-L2, FR-L3 located between CDR-L2 and CDR-L3 and so forth. It is noted that because the shown Kabat numbering scheme places insertions at H35A and H35B, the end of the Chothia CDR-H1 loop when numbered using the shown Kabat numbering convention varies between H32 and H34, depending on the length of the loop.Table 1 Boundaries of CDRs according to various numbering schemes1 - 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

[0297] Thus, unless otherwise specified, a “CDR” or “complementary determining region,” or individual specified CDRs ( e.g ., CDR-H1, CDR-H2, CDR-H3), of a given antibody or region thereof, such as a variable region thereof, should be understood to encompass a (or the specific) complementary determining region as defined by any of the aforementioned schemes, or other known schemes. For example, where it is stated that a particular CDR (e.g., a CDR-H3) contains the amino acid sequence of a corresponding CDR in a given VH or VL region amino acid sequence, it is understood that such a CDR has a sequence of the corresponding CDR (e.g., CDR-H3) within the variable region, as defined by any of the aforementioned schemes, or other known schemes. In some embodiments, specific CDR sequences are specified. Exemplary CDR sequences of provided antibodies are described using various numbering schemes, although it is understood that a provided antibody can include CDRs as described according to any of the other aforementioned numbering schemes or other numbering schemes known to a skilled artisan.

[0298] Likewise, unless otherwise specified, a FR or individual specified FR(s) (e.g., FR- Hl, FR-H2, FR-H3, FR-H4), of a given antibody or region thereof, such as a variable region thereof, should be understood to encompass a (or the specific) framework region as defined by any of the known schemes. In some instances, the scheme for identification of aparticular CDR, FR, or FRs or CDRs is specified, such as the CDR as defined by the Kabat, Chothia, AbM, IMGT or Contact method, or other known schemes. In other cases, the particular amino acid sequence of a CDR or FR is given.

[0299] 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 antigen. The variable regions of the heavy chain and light chain (VH and VL, respectively) of a native antibody generally have similar structures, with each domain comprising 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 VH or VL domain may be sufficient to confer a capability for binding an antigen. Furthermore, antibodies that bind a particular antigen may be isolated using a VH or VL domain from an antibody that binds the antigen to screen a library of complementary VL or VH domains, respectively. See, e.g., Portolano et ah, J. Immunol. 150:880-887 (1993); Clarkson et al, Nature 352:624-628 (1991).

[0300] An “antibody fragment” or “antigen-binding fragment” refers to a molecule other than an intact antibody that comprises a portion of an intact antibody that is capable of binding 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 (VH) regions, single-chain antibody molecules such as scFvs and single-domain antibodies comprising only the VH region; and multispecific antibodies formed from antibody fragments. In particular embodiments, the antibodies from which the CARs are derived are single-chain antibody fragments comprising a heavy chain variable (VH) region and / or a light chain variable (VL) region, such as scFvs.

[0301] Single-domain antibodies (sdAbs) are antibody fragments comprising 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, a single-domain antibody is a human singledomain antibody.

[0302] 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 optionally may include 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 the non-human antibody that has undergone humanization, typically to reduce immunogenicity to humans, while retaining the specificity and affinity of the parental non-human antibody. In some embodiments, some FRresidues in a humanized antibody are substituted with corresponding residues from a nonhuman antibody (e.g the antibody from which the CDR residues are derived), e.g., to restore or improve antibody specificity or affinity.

[0303] A “human antibody” is an antibody with an amino acid sequence corresponding to that of an antibody produced by a human or a human cell, or non-human source that utilizes human antibody repertoires or other human antibody-encoding sequences, including human antibody libraries. The term excludes humanized forms of non-human antibodies comprising non-human antigen-binding regions, such as those in which all or substantially all CDRs are non-human. The term includes antigen-binding fragments of human antibodies. Human antibodies may be prepared by administering an immunogen to a transgenic animal that has 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 a portion of the human immunoglobulin loci, which replace the endogenous immunoglobulin loci, or which are present extrachromosomally or integrated randomly into the animal’s chromosomes. In such transgenic animals, the endogenous immunoglobulin loci have generally been inactivated. Human antibodies also may be derived from human antibody libraries, including phage display and cell-free libraries, containing antibody-encoding sequences derived from a human repertoire.

[0304] The term “monoclonal antibody” as used herein refers to an antibody obtained from or within a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical, except for possible variants containing naturally occurring mutations or arising during production of a monoclonal antibody preparation, such variants generally being present in minor amounts. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different epitopes, each monoclonal antibody of a monoclonal antibody preparation is directed against a single epitope on an antigen. The term is not to be construed as requiring production of the antibody by any particular method. A monoclonal antibody may be made by a variety of techniques, including but not limited to generation from a hybridoma, recombinant DNA methods, phage-display and other antibody display methods.

[0305] In some embodiments, the binding (e.g., antibody-antigen binding or receptorantigen binding) is “specific.” For example, in some embodiments, the monoclonal antibody of the disclosure is capable of specifically binding to the peptide epitope or mimotope. In some embodiments, the receptor or the binding region of the disclosure is capable ofspecifically binding to the antigen. As used herein, the terms “specific” or “specifically,” when used with reference to binding, refer to measurable and reproducible interactions, such as binding between a target and an antibody or scFv or binding region or receptor, which is determinative of the presence of the target in the presence of a heterogeneous population of molecules including biological molecules. For example, an antibody, scFv, binding region, or receptor that specifically binds to a target (which can be an epitope or an antigen) is an antibody, scFv, binding region, or receptor that binds this target with greater affinity, avidity, more readily, and / or with greater duration than it binds to other molecules (e.g., the extent of binding of the antibody, scFv, binding region, or receptor to an unrelated molecule is less than about 10% of the binding of the antibody, scFv, binding region, or receptor to the target as measured, e.g., by surface plasmon resonance (SPR)). In some embodiments, a specific binding has a dissociation constant (Kd) of < 1 pM, < 100 nM, < 10 nM, < 1 nM, or < 0.1 nM. In some embodiments, specific binding can include, but does not require, exclusive binding. In some embodiments, the specific binding is exclusive binding.

[0306] The term “engineered cell” as used herein refers to a cell that has been altered in at least some way by human intervention, including, for example, by genetic alterations or modifications such that the engineered cell differs from a wild-type cell.

[0307] The terms “decrease,” “reduced,” “reduction,” and “decreased” are all used herein generally to mean a lowering by a statistically significant amount. However, for avoidance of doubt, “decrease,” “reduced,” “reduction,” “decreased” means a lowering by at least 10% as compared to a reference level, for example a lowering by at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90%, or up to and including a 100% lowering (i.e. absent level as compared to a reference sample), or any lowering between 10- 100% as compared to a reference level. In some embodiments, the cells are engineered to have reduced expression of one or more targets relative to an unaltered or unmodified wildtype cell.

[0308] The terms “increase” and “increased” are all used herein generally to mean an increase by a statistically significant amount. However, for avoidance of doubt, “increase” and “increased” mean an increase by at least 10% as compared to a reference level, for example an increase by at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least 1-fold, at least 2-fold, at least 3-fold, at least 5-fold, at least 10-fold, at least 20-fold, atleast 30-fold, at least 50-fold, at least 100-fold, at least 200-fold, at least 500-fold, or at least 1000-fold, or any increase between 10% and 1000-fold as compared to a reference level. In some embodiments, the cells are engineered to have increased expression of one or more targets relative to an unaltered or unmodified wild-type cell.

[0309] As used herein, the term “mimotope” refers to a molecule, or a part of a biomolecule, that mimics the structure of an epitope and is capable of being bound by the same antigen-binding region of an antibody that recognize the epitope. The mimotope may act as a competitor for the corresponding epitope in in vitro assays (e.g., ELISA) and / or elicit an immunological response in a host that is reactive to the epitope of which it is a mimic. In some embodiments, the mimotope of the disclosure is a peptide.

[0310] As used herein, the terms “conservative amino acid substitution” and “conservative substitution” refer to amino acid substitutions in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine).

[0311] Headings, e.g., (a), (b), (i) etc., are presented merely for ease of reading the specification and claims. The use of headings in the specification or claims does not require the steps or elements be performed in alphabetical or numerical order or the order in which they are presented.lt is appreciated that certain features of the disclosure, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the disclosure, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable sub-combination. All combinations of the embodiments pertaining to the disclosure are specifically embraced by the present disclosure and are disclosed herein just as if each and every combination was individually and explicitly disclosed. In addition, all sub-combinations of the various embodiments and elementsthereof are also specifically embraced by the present disclosure and are disclosed herein just as if each and every such sub- combination was individually and explicitly disclosed herein.IL Epitope or Mimitope Tag

[0312] In one aspect, the disclosure provides an epitope or mimotope. In some embodiments, the epitope or mimotope of the disclosure can be used as depletion tag on the surface of engineered immune cells (e.g., CAR-T cells) to improve the clinical safety of those cells.

[0313] In some embodiments, the epitope or mimotope is comprised within a recombinant polypeptide of the disclosure. In some embodiments, the epitope or mimotope is located in the extracellular region of the recombinant polypeptide. In some embodiments, the recombinant polypeptide can be expressed on the surface of an engineered cell of the disclosure. In some embodiments, the recombinant polypeptide comprises a transmembrane region. In some embodiments, the recombinant polypeptide comprises a glycosylphosphatidylinositol (GPI) anchor. In some embodiments, the recombinant polypeptide comprises a signal peptide during protein translation that enables its expression on the cell surface. In some embodiments, such cell surface expression presents the epitope or mimotope tag so that it is capable of being bound by a corresponding antibody for the epitope or mimotope.

[0314] In some embodiments, the epitope or a mimotope tag is comprised within a recombinant receptor of the disclosure.

[0315] In some embodiments, the peptide epitope is derived from an autoimmune- associated antigen. In some embodiments, the peptide epitope is derived from a cancer- associated antigen.

[0316] In some embodiments, the mimotope mimics an epitope of an autoimmune- associated antigen that is bound by a monoclonal antibody. In some embodiments, the mimotope mimics an epitope of a cancer-associated antigen that is bound by a monoclonal antibody.

[0317] In some embodiments, the autoimmune-associated antigen or cancer-associated antigen is selected from the group consisting of CD1, CD la, CD lb, CDlc, CD Id, CDle, CD2, CD3d, CD3e, CD3g, CD3s, CD4, CD5, CD7, CD8a, CD8b, CD19, CD20, CD21, CD22, CD23, CD24, CD25, CD27, CD28, CD30, CD33, CD34, CD38, CD40, CD44v6, CD45, CD46, CD47 CD48, CD52, CD59, CD66, CD70, CD71, CD72, CD73, CD79A,CD79B, CD80 (B7.1), CD86 (B7.2), CD94, CD95, CD97, CD123, CD134, CD140 (PDGFR4), CD152, CD154, CD158, CD171, CD178, CD179, CD179a, CD181 (CXCR1), CD182 (CXCR2), CD183 (CXCR3), CD210, CD213A2, CD246, CD252, CD253, CD261, CD262, CD272, CD273 (PD-L2), CD274 (PD-L1), CD276 (B7H3), CD279, CD295, CD339 (JAG1), CD340 (HER2), CDH17, CEA, CLECL1, CLL-1, CLDN6, CLDN18.2, CS1, DLL3, LY6G6D, GCC, p53R175H, PRAME, EGFR, EGFRvIII, FGFR2, AFP, CA125, MUC-1, MAGE, ALPI, alkaline phosphatase placental-like 2 (ALPPL2), B-cell maturation antigen (BCM A), green fluorescent protein (GFP), enhanced green fluorescent protein (eGFP), KLK2, KLK3, Mesothelin, IL13Ra2, signal regulatory protein a (SIRPa), TCRalpha, TCRbeta, TSHR, GD2, GD3, Tn Ag, cMET, Axl, R0R1, R0R2, GPC1, GPC2, GPC3, FLT3, TAG72, CEA, EPC AM, KIT (CD117), IL-13Ra2, IL-l lRa, PSCA, PRSS21, VEGFR2, LewisY, PDGFRp, SSEA-4, folate receptor alpha, ERBB2 (Her2 / neu), MUC1, MUC16, NCAM, prostase, PAP, ELF2M, Ephrin B2, IGF-I receptor, CAIX, LMP2, gplOO, bcr-abl, tyrosinase, EphA2, STEAP1, STEAP2, fucosyl GM1, sLe, GM3, TGS5, HMWMAA, o-acetyl-GD2, folate receptor beta, TEM1 / CD248, TEM7R, R0PN1, GPRC5D, GPA33, CX0RF61, ALK, Polysialic acid, PLAC1, GloboH, NY-BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP, WT1, NY-ESO-1, LAGE-la, MAGE-A1, legumain, HPV E6,E7, MAGE Al, ETV6-AML, sperm protein 17, XAGE1, Tie 2, MAD-CT-1, MAD-CT-2, Fos-related antigen 1, p53, p53 mutant, p53R175H, KRAS, mutant KRAS, KRAS G12D, prostein, surviving, telomerase, PCTA-l / Galectin 8, MelanA / MARTl, Ras mutant, hTERT, sarcoma translocation breakpoints, ML-IAP, ERG (TMPRSS2 ETS fusion gene), NA17, PAX3, androgen receptor, cyclin Bl, MYCN, RhoC, TRP-2, CYP1B1, BORIS, SART3, PAX5, OY-TES1, LCK, AKAP-4, SSX2, RAGE-1, human telomerase reverse transcriptase, RU1, RU2, intestinal carboxyl esterase, mut hsp70- 2,LAIR1, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, FCRL5, IGLL1, PSMA, TROP2, citrullinated vimentin, the extracellular portion of the APRIL protein, and any combinations thereof.

[0318] In some embodiments, the cancer-associated antigen is a cancer-ssociated antigen for a hematologic cancer. In some embodiments, the cancer is selected from B cell lymphoma, T cell lymphoma, chronic lymphocytic leukemia, and acute myeloid leukemia.

[0319] In some embodiments, the cancer-associated antigen is a cancer-associated antigen for a solid tumor. In some embodiments, the cancer is a solid tumor selected from the group consisting of small cell lung cancer, prostate cancer, colorectal cancer, head and necksquamous cell carcinoma, breast cancer, melanoma, ovarian cancer, lung cancer, pancreatic cancer, stomach cancer, cervical cancer, hepatocellular carcinoma, glioma cancer, neuroblastoma cancer, and renal cell carcinoma. In some embodiments, the cancer- associated antigen for the solid tumor comprises or consists of KIR2D, NRP1, CD221, VEGFR, HGF, integrin a5 1, CD51, CD27, B7H3, NYESO1, EGFR, cMET, BRD4, EpCAM, SSX, Fibronectin EDB, integrin al ipi, DLL3, CD2, SSTR2, TROP2, KLK2, KLK3, STEAP2, STEAP1, PSMA, PSCA, LY6G6D, IL-la, CEA, C242 antigen, CanAg (MUC1 glycoform), CTAA16.88, UCY2C, IGF2BP3, p53R175H, NKG2D ligands, PRAME, MAGEA4, vimentin, HER2 / neu, CCR4, CCR5, R0PN1, R0R1, KK-LC-1, CA- 125, folate receptor 1, folate receptor alpha, MUC1, MUC16, MSLN, CLDN6, WT1, ALPPL2, KRASG12D, KRASG12V, CD70, MSLN, CLDN18.2, HPV E6 / E7, GPC3, IL- 13Ra, EGFRv3, EGFR806, GD2, or CA9. In some embodiments, the cancer-associated antigen for the solid tumor comprises or consists of KIR2D, NRP1, CD221, VEGFR, HGF, integrin a5bl, CD51, CD27, B7H3, NYESO1, EGFR, cMET, BRD4, EpCAM, SSX, Fibronectin EDB, or Integrin al ipi. In some embodiments, the cancer-associated antigen for the solid tumor comprises or consists of KLK2, Ly6G6D, CLDN6, DLL3 or CD 19.

[0320] In some embodiments, the cancer-associated antigen is selected from one listed in Table 2 below.Table 2. Cancers and Associated Antigens

[0321] In some embodiments, the antigen is not CD 19. In some embodiments, the antigen is not CCR4. In some embodiments, the antigen is not CCR7.

[0322] In some embodiments, the antigen is Delta-like protein 3 (DLL3). Human DLL3 (Uniprot Accession No. Q9NYJ7) is a biomarker for small cell lung cancer. In some embodiments, the DLL3 protein comprises an amino acid sequence at least at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 486. In some embodiments, the epitope comprises at least 10, at least 15, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, or at least 50, consecutive amino acids in SEQ ID NO: 486, or an amino acid sequence having at most 1, at most 2, at most 3, at most 4, or at most 5 amino acid mutations thereto. In some embodiments, the epitope comprises about 10 to about 15, about 15 to about 20, about 20 to about 25, about 25 to about 30, about 30 to about 35, about 35 to about 40, about 40 to about 45, or about 45 to about 50, consecutive amino acids in SEQ ID NO: 486, or an amino acid sequence having at most 1, at most 2, at most 3, at most 4, or at most 5 amino acid mutations thereto. In some embodiments, the peptide epitope comprises or consists of any one of SEQ ID NO: 439-468, or an amino acid sequence having at most 1, at most 2, at most 3, at most 4, or at most 5 amino acid mutations thereto. In some embodiments, the peptide epitope comprises or consists of SEQ ID NO: 439, or an amino acid sequence having at most 1, atmost 2, at most 3, at most 4, or at most 5 amino acid mutations thereto. In some embodiments, the peptide epitope comprises or consists of SEQ ID NO: 439.

[0323] In some embodiments, the epitope is derived from a cancer-associated antigen of adult T-cell lymphoma (ATL) and / or cutaneous T-cell lymphoma (CTCL). In some embodiments, the epitope is derived from a cancer-associated antigen of a Th2 cell, a cutaneous lymphocyte antigen-positive skin-homing T cell, and / or a Treg cell.

[0324] In some embodiments, the epitope is derived from C-C chemokine receptor type 4 (CCR4) (i.e., the cancer associated antigen is CCR4). C-C chemokine receptor type 4 (CCR4), also known as CD 194, is a biomarker for T cell lymphoma. Further information of human CCR4 protein is described in Uniprot Accession No. P51679. In some embodiments, the CCR4 protein comprises an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 485. In some embodiments, the epitope comprises at least 10, at least 15, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, or at least 50, consecutive amino acids in SEQ ID NO: 485, or an amino acid sequence having at most 1, at most 2, at most 3, at most 4, or at most 5 amino acid mutations thereto. In some embodiments, the epitope comprises about 10 to about 15, about 15 to about 20, about 20 to about 25, about 25 to about 30, about 30 to about 35, about 35 to about 40, about 40 to about 45, or about 45 to about 50, consecutive amino acids in SEQ ID NO: 485, or an amino acid sequence having at most 1, at most 2, at most 3, at most 4, or at most 5 amino acid mutations thereto. In some embodiments, the peptide epitope comprises or consists of an amino acid sequence of any one of SEQ ID NO: 430-438, or an amino acid sequence having at most 1, at most 2, at most 3, at most 4, or at most 5 amino acid mutations thereto. In some embodiments, the peptide epitope comprises or consists of an amino acid sequence of SEQ ID NO: 430. In some embodiments, the peptide epitope comprises or consists of an amino acid sequence of SEQ ID NO: 431.

[0325] In some embodiments, the epitope is derived from C-C chemokine receptor type 7 (CCR7) (i.e., the cancer associated antigen is CCR7). C-C chemokine receptor type 7 (CCR7), also known as CD 197, is a biomarker for types of cancer including breast, lung, pancreatic, squamous cell carcinoma, and gastric cancer. CCR7 is also associated with cancer progression and poor outcomes, including metastasis. Further information of human CCR7 protein is described in Uniprot Accession No. P32248. In some embodiments, the CCR7 protein comprises an amino acid sequence at least 80%, at least 85%, at least 90%, atleast 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 557. In some embodiments, the epitope comprises at least 10, at least 15, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, or at least 50, consecutive amino acids in SEQ ID NO: 557, or an amino acid sequence having at most 1, at most 2, at most 3, at most 4, or at most 5 amino acid mutations thereto. In some embodiments, the epitope comprises about 10 to about 15, about 15 to about 20, about 20 to about 25, about 25 to about 30, about 30 to about 35, about 35 to about 40, about 40 to about 45, or about 45 to about 50, consecutive amino acids in SEQ ID NO: 557, or an amino acid sequence having at most 1, at most 2, at most 3, at most 4, or at most 5 amino acid mutations thereto. In some embodiments, the peptide epitope comprises or consists of an amino acid sequence of any one of SEQ ID NO: 519-524, or an amino acid sequence having at most 1, at most 2, at most 3, at most 4, or at most 5 amino acid mutations thereto. In some embodiments, the peptide epitope comprises or consists of an amino acid sequence of SEQ ID NO: 519. In some embodiments, the peptide epitope comprises or consists of an amino acid sequence of SEQ ID NO: 520. In some embodiments, the peptide epitope comprises or consists of an amino acid sequence of SEQ ID NO: 521. In some embodiments, the peptide epitope comprises or consists of an amino acid sequence of SEQ ID NO: 522. In some embodiments, the peptide epitope comprises or consists of an amino acid sequence of SEQ ID NO: 523. In some embodiments, the peptide epitope comprises or consists of an amino acid sequence of SEQ ID NO: 524.

[0326] In some embodiments, the peptide epitope or the mimotope is about 5 to about 10, about 10 to about 20, about 15 to about 25, about 20 to about 30, about 25 to about 35, about 30 to about 40, about 35 to about 45, or about 40 to about 50 amino acids in length. In some embodiments, the peptide epitope or the mimotope is no more than 10, no more than 15, no more than 20, no more than 25, no more than 30, no more than 35, no more than 40, no more than 45, or no more than 50 amino acids in length. In some embodiments, the peptide epitope does not comprise any mutation compared to the native epitope derived from the antigen. In some embodiments, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% of the peptide epitope or the mimotope is capable of forming an alpha-helical conformation.

[0327] In some embodiments, the epitope or mimotope is capable of being bound by the antigen-binding molecule (e.g., antibody) with a dissociation constant (Kd) of less than 100 pM, less than 10 pM, less than 1 pM, less than 100 nM, less than 10 nM, less than 1 nM,less than 100 pM, less than 10 pM, or less than 1 pM, optionally wherein the Kd is measured by surface plasmon resonance (SPR) using a sensor chip that contains the immobilized the antigen-binding molecule (e.g., antibody). In some embodiments, the modified peptide epitope or mimotope is capable of being bound by the antigen-binding molecule (e.g., antibody) with a dissociation constant (Kd) of about 100 pM to about 10 pM, about 10 pM to about 1 pM, about 1 pM to about 100 nM, about 100 nM to about 10 nM, about 10 nM to about 1 nM, about 1 nM to about 100 pM, about 100 pM to about 10 pM, or about 10 pM to about 1 pM, optionally wherein the Kd is measured by surface plasmon resonance (SPR) using a sensor chip that contains the immobilized antigen-binding molecule (e.g., antibody).

[0328] In some embodiments, the epitope or mimotope is that of an antibody. In some embodiments, the antibody is [fam-]trastuzumab deruxtecan, (fam-trastuzumab deruxtecan- nxki), Adebrelimab, Ado-trastuzumab emtansine, Alemtuzumab, Amivantamab, Atezolizumab, Avelumab, Belantamab mafodotin (belantamab mafodotin-blmf), Bevacizumab, Blinatumomab, Brentuximab vedotin, Cadonilimab, Camrelizumab, CAP- 100, Catumaxomab, Cemiplimab (cemiplimab-rwlc), Cetuximab, Cetuximab saratolacan, Cosibelimab, Daratumumab, Dinutuximab, Disitamab vedotin, Dostarlimab, Durvalumab, Edrecolomab, Elotuzumab, Elranatamab, Emapalumab (emapalumab-lzsg), Enfortumab vedotin (enfortumab vedotin-ejfv), Enlonstobart, Envafolimab, Epcoritamab, Gemtuzumab ozogamicin, Glofitamab, Ibritumomab tiuxetan, Inotuzumab ozogamicin, Ipilimumab, Isatuximab (isatuximab-irfc), Loncastuximab tesirine, Margetuximab-cmkb, Mirvetuximab soravtansine, Mogamulizumab (mogamulizumab-kpkc), Mosunetuzumab, Moxetumomab pasudotox (moxetumomab pasudotox-tdfk), Naxitamab-gqgk, Necitumumab, Nimotuzumab, Nivolumab, Cipterbin, Obinutuzumab, Ofatumumab, Olaratumab, Panitumumab, Pembrolizumab, Penpulimab, Pertuzumab, Polatuzumab vedotin (polatuzumab vedotin- piiq), Prolgolimab, Pucotenlimab, Racotumomab, Ramucirumab, Relatlimab, Retifanlimab, Ripertamab, Rituximab, Sacituzumab govitecan (sacituzumab govitecan-hziy), Serplulimab, Sintilimab, Socazolimab, Sugemalimab, Tafasitamab (tafasitamab-cxix), Tagitanlimab, Talquetamab (talquetamab-tgvs), Tebentafusp, Teclistamab, Tislelizumab, Tisotumab vedotin, tisotumab vedotin-tftv, Toripalimab, Tositumomab-1131, Trastuzumab, Trastuzumab duocarmazine, Tremelimumab, Zimberelimab, Zolbetuximab, Zuberitamab, Odronextamab, Ivonescimab, Benmelstobart, Trastuzumab botidotin, Iparomlimab, tuvonralimab, Tarlatamab, Patritumab deruxtecan, Sacituzumab tirumotecan, Zanidatamab, Linvoseltamab, Datopotamab deruxtecan, Anbenitamab, Anvatabart opadotin,Apamistamab-Iodine (1311), Bemarituzumab, Cetrelimab, Cobolimab, Domvanalimab, Emactuzumab, Erfonrilimab, Favezelimab, Felzartamab, Fianlimab, Finotonlimab, ABBV- 383, Geptanolimab, Gotistobart, Ivuxolimab, Izalontamab, Lemzoparlimab, Luveltamab tazevibulin, Magrolimab, Mecbotamab vedotin, Monalizumab, Nofazinlimab, Nurulimab, Ociperlimab, Oleclumab, Onfekafusp alfa, Oregovomab, Ozekibart, Pivekimab sunirine, Quavonlimab, Retlirafusp alfa, Rosopatamab (177Lu-DOTA), Rulonilimab, Sasanlimab, Telisotuzumab vedotin, Tiragolumab, IKS014, LCB14-0110, FS-1502, Trastuzumab mafodotin, Trastuzumab rezetecan, Vibostolimab, Vobramitamab duocarmazine, Zilovertamab vedotin, Suvemcitug, APX003, BD0801, TK001, sevacizumab, CTX-009, ES104, TR009, NOV1501, ABL001, DP303c, IAH0968, Kintuximab, gentuximab, cintuximab, MIL62, MRG002, MRG003, PM8002, MEDI5752, MCLA-128, AGEN1181, L19-IL2, L19-TNF, IMC-F106C, JMT101, BL-B01D1, BAT1308, AZD2936, JS004, TAB004, FG-M108, M108, 9MW2821, BMS-986349, CC-93269, EM801, ABBV-181, ABBV-151, ARGX-115, DS-7300a, IBI343, SHR-A1921, or ASKB589. In some embodiments, the antibody is tarlatamab. In some embodiments, the antibody is mogamulizumab. In some embodiments, the antibody is CAP-100.Modified Epitope or Mimotope

[0329] In some embodiments, the peptide epitope or mimotope of the disclosure comprises or consists of an amino acid sequence having at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mutations compared to an unmodified peptide epitope or mimotope and is capable of being bound by the corresponding monoclonal antibody that the unmodified epitope or mimotope binds to. In some embodiments, the peptide epitope or mimotope comprises or consists of an amino acid sequence having 1, 2, 3, 4, or 5 mutations compared to the unmodified peptide epitope or mimotope. In some embodiments, the peptide epitope or mimotope comprises or consists of an amino acid sequence having 5 mutations compared to the unmodified peptide epitope or mimotope. In some embodiments, the peptide epitope or mimotope comprises or consists of an amino acid sequence having 4 mutations compared to the unmodified peptide epitope or mimotope. In some embodiments, the peptide epitope or mimotope comprises or consists of an amino acid sequence having 3 mutations compared to the unmodified peptide epitope or mimotope. In some embodiments, the peptide epitope or mimotope comprises or consists of an amino acid sequence having 2 mutations compared to the unmodified peptide epitope or mimotope. In some embodiments, the peptide epitope or mimotope comprises or consistsof an amino acid sequence having 1 mutation compared to the unmodified peptide epitope or mimotope.

[0330] In some embodiments, the antibody binding affinity of the modified epitope or mimotope is at a comparable level as the antibody binding affinity of the unmodified epitope or mimotope. In some embodiments, the modified peptide epitope or mimotope is capable of being bound by the corresponding antibody with a dissociation constant (Kd) of less than 100 pM, less than 10 pM, less than 1 pM, less than 100 nM, less than 10 nM, less than 1 nM, less than 100 pM, less than 10 pM, or less than 1 pM, optionally wherein the Kd is measured by surface plasmon resonance (SPR) using a sensor chip that contains the immobilized antibody. In some embodiments, the modified peptide epitope or mimotope is capable of being bound by the corresponding antibody with a dissociation constant (Kd) of about 100 pM to about 10 pM, about 10 pM to about 1 pM, about 1 pM to about 100 nM, about 100 nM to about 10 nM, about 10 nM to about 1 nM, about 1 nM to about 100 pM, about 100 pM to about 10 pM, or about 10 pM to about 1 pM, optionally wherein the Kd is measured by surface plasmon resonance (SPR) using a sensor chip that contains the immobilized antibody.

[0331] In some embodiments, the peptide epitope or mimotope of the disclosure comprises or consists of an amino acid sequence having no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mutations compared to an unmodified peptide epitope or mimotope and is capable of being bound by the corresponding monoclonal antibody that the unmodified epitope or mimotope binds to.

[0332] In some embodiments, the peptide epitope or mimotope of the disclosure comprises or consists of an amino acid sequence having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mutations compared to an unmodified peptide epitope or mimotope and is capable of being bound by the corresponding monoclonal antibody that the unmodified epitope or mimotope binds to. In some embodiments, the peptide epitope or mimotope of the disclosure comprises or consists of an amino acid sequence having 1-2, 2-3, 3-4, 4-5, 5-6, 6-7, 7-8, 8-9, or 9-10 mutations compared to an unmodified peptide epitope or mimotope and is capable of being bound by the corresponding monoclonal antibody that the unmodified epitope or mimotope binds to.

[0333] In some embodiments, the one or more mutations in the modified epitope or mimotope comprise or consist of substitutions, insertions, deletions, or any combinationthereof. In some embodiments, the one or more mutations in the modified epitope or mimotope comprise or consist of substitutions and / or deletions.

[0334] In some embodiments, the one or more mutations in the modified epitope or mimotope comprise or consist of substitutions. In some embodiments, the substitution(s) are conservative amino acid substitutions.

[0335] In some embodiments, the mutation comprises addition and / or deletion of one or more amino acids at the N-term or C-term of the unmodified epitope or mimotope. In some embodiments, the mutation comprises deleting 1, 2, 3, 4, or 5, amino acids at the N-terminus of the unmodified epitope or mimotope. In some embodiments, the mutation comprises deleting 1, 2, 3, 4, or 5, amino acids at the C-terminus of the unmodified epitope or mimotope. In some embodiments, the mutation comprises inserting 1, 2, 3, 4, or 5, amino acids at the N-terminus of the unmodified epitope or mimotope. In some embodiments, the mutation comprises inserting 1, 2, 3, 4, or 5, amino acids at the C-terminus of the unmodified epitope or mimotope. In some embodiments, the mutation comprises deleting 1, 2, 3, 4, or 5, amino acids at the N-terminus of the unmodified epitope or mimotope and inserting 1, 2, 3, 4, or 5, amino acids at the C-terminus of the unmodified epitope or mimotope. In some embodiments, the mutation comprises inserting 1, 2, 3, 4, or 5, amino acids at the N-terminus of the unmodified epitope or mimotope and deleting 1, 2, 3, 4, or 5, amino acids at the C-terminus of the unmodified epitope or mimotope.

[0336] In some embodiments, the cysteine(s) in the unmodified peptide epitope or mimotope are substituted. In some embodiments, the N-terminus of the unmodified peptide epitope or mimotope contains cysteine. In some embodiments, the C-terminus of the unmodified peptide epitope or mimotope contains cysteine. In some embodiments, the unmodified peptide epitope or mimotope comprises at least one cysteine between the N- terminus and the C-terminus. In some embodiments, the cysteine(s) are substituted with serine, glycine, threonine, alanine, valine, or any combination thereof. In some embodiments, the cysteine(s) are substituted with serine and / or valine. In some embodiments, the cysteine(s) are substituted with serine(s). In some embodiments, the cysteine(s) are substituted with valine(s). In some embodiments, the modified epitope or mimotope does not comprise any cysteine(s).

[0337] In some embodiments, the mutation(s) comprise deleting or substituting one or more amino acids that facilitate polypeptide crosslinking in the unmodified peptide epitope. In some embodiments, the mutation(s) comprise deleting and / or substituting one or morecysteine(s), lysine(s), aspartic acid(s), glutamic acid(s), or any combination thereof, in the unmodified peptide epitope or mimotope.

[0338] In some embodiments, the unmodified peptide epitope or mimotope comprises or consists of no more than 10, no more than 15, no more than 20, no more than 25, no more than 30, no more than 35, no more than 40, no more than 45, or no more than 50, consecutive amino acids in an autoimmune-associated antigen or a cancer-associated antigen. In some embodiments, the unmodified peptide epitope or mimotope comprises or consists of about 10 to about 15, about 15 to about 20, about 20 to about 25, about 25 to about 30, about 30 to about 35, about 35 to about 40, about 40 to about 45, or about 45 to about 50, consecutive amino acids in an autoimmune-associated antigen or a cancer- associated antigen.

[0339] In some embodiments, the epitope or mimotope is capable of being bound by a monoclonal antibody that targets a biomarker for a hematologic cancer. In some embodiments, the epitope or mimotope is capable of being bound by an anti-CCR4 monoclonal antibody. In some embodiments, the CCR4 protein comprises an amino acid sequence at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 485. In some embodiments, the unmodified epitope or mimotope comprises at least 10, at least 15, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, or at least 50, consecutive amino acids in SEQ ID NO: 485. In some embodiments, the unmodified epitope or mimotope comprises about 10 to about 15, about 15 to about 20, about 20 to about 25, about 25 to about 30, about 30 to about 35, about 35 to about 40, about 40 to about 45, or about 45 to about 50, consecutive amino acids in SEQ ID NO: 485.

[0340] In some embodiments, the epitope or mimotope is capable of being bound by mogamulizumab, a humanized, afucosylated monoclonal antibody targeting CCR4. The unmodified epitope of mogamulizumab has the amino acid sequence of DESIYSNYYLYESIPKPC (SEQ ID NO: 430). In some embodiments, the modified epitope comprising a substitution or deletion of the cysteine at the C-term of SEQ ID NO: 430. In some embodiments, the modified epitope comprises a serine, glycine, threonine, alanine, or valine substitution of the cysteine at the C-term of SEQ ID NO: 430. In some embodiments, the modified epitope comprises or consists of any one of SEQ ID NO: 432-436. In some embodiments, the modified epitope comprises the deletion of 1, 2, 3, 4, or 5 amino acids at the C-term of SEQ ID NO: 430. In some embodiments, the modified epitope comprises orconsists of SEQ ID NO: 431. . In some embodiments, the modified epitope comprises or consists of SEQ ID NO: 438. In some embodiments, the modified epitope comprises the insertion of 1, 2, 3, 4, or 5, amino acid from the corresponding position of CCR4 (SEQ ID NO: 485) before the N-term of SEQ ID NO: 430. In some embodiments, the modified epitope comprises or consists of SEQ ID NO: 437.

[0341] In some embodiments, the modified peptide epitope or mimotope comprises or consists of an amino acid sequence having 1, 2, 3, 4 or 5 mutations to any one of the unmodified epitopes and mimotopes listed in Table 3 below and is capable of being bound by the corresponding monoclonal antibody that binds to that unmodified epitope or mimotope.Table 3. Epitopes and Mimotopes

[0342] In some embodiments, the modified peptide epitope or mimotope is capable of being bound by [fam-]trastuzumab deruxtecan, (fam-trastuzumab deruxtecan-nxki), Adebrelimab, Ado-trastuzumab emtansine, Alemtuzumab, Amivantamab, Atezolizumab, Avelumab, Belantamab mafodotin (belantamab mafodotin-blmf), Bevacizumab, Blinatumomab, Brentuximab vedotin, Cadonilimab, Camrelizumab, CAP- 100, Catumaxomab, Cemiplimab (cemiplimab-rwlc), Cetuximab, Cetuximab saratolacan, Cosibelimab, Daratumumab, Dinutuximab, Disitamab vedotin, Dostarlimab, Durvalumab, Edrecolomab, Elotuzumab, Elranatamab, Emapalumab (emapalumab-lzsg), Enfortumab vedotin (enfortumab vedotin-ejfv), Enlonstobart, Envafolimab, Epcoritamab, Gemtuzumab ozogamicin, Glofitamab, Ibritumomab tiuxetan, Inotuzumab ozogamicin, Ipilimumab,Isatuximab (isatuximab-irfc), Loncastuximab tesirine, Margetuximab-cmkb, Mirvetuximab soravtansine, Mogamulizumab (mogamulizumab-kpkc), Mosunetuzumab, Moxetumomab pasudotox (moxetumomab pasudotox-tdfk), Naxitamab-gqgk, Necitumumab, Nimotuzumab, Nivolumab, Cipterbin, Obinutuzumab, Ofatumumab, Olaratumab, Panitumumab, Pembrolizumab, Penpulimab, Pertuzumab, Polatuzumab vedotin (polatuzumab vedotin- piiq), Prolgolimab, Pucotenlimab, Racotumomab, Ramucirumab, Relatlimab, Retifanlimab, Ripertamab, Rituximab, Sacituzumab govitecan (sacituzumab govitecan-hziy), Serplulimab, Sintilimab, Socazolimab, Sugemalimab, Tafasitamab (tafasitamab-cxix), Tagitanlimab, Talquetamab (talquetamab-tgvs), Tebentafusp, Teclistamab, Tislelizumab, Tisotumab vedotin, tisotumab vedotin-tftv, Toripalimab, Tositumomab-1131, Trastuzumab, Trastuzumab duocarmazine, Tremelimumab, Zimberelimab, Zolbetuximab, Zuberitamab, Odronextamab, Ivonescimab, Benmelstobart, Trastuzumab botidotin, Iparomlimab, tuvonralimab, Tarlatamab, Patritumab deruxtecan, Sacituzumab tirumotecan, Zanidatamab, Linvoseltamab, Datopotamab deruxtecan, Anbenitamab, Anvatabart opadotin, Apamistamab-Iodine (1311), Bemarituzumab, Cetrelimab, Cobolimab, Domvanalimab, Emactuzumab, Erfonrilimab, Favezelimab, Felzartamab, Fianlimab, Finotonlimab, ABBV- 383, Geptanolimab, Gotistobart, Ivuxolimab, Izalontamab, Lemzoparlimab, Luveltamab tazevibulin, Magrolimab, Mecbotamab vedotin, Monalizumab, Nofazinlimab, Nurulimab, Ociperlimab, Oleclumab, Onfekafusp alfa, Oregovomab, Ozekibart, Pivekimab sunirine, Quavonlimab, Retlirafusp alfa, Rosopatamab (177Lu-DOTA), Rulonilimab, Sasanlimab, Telisotuzumab vedotin, Tiragolumab, IKS014, LCB14-0110, FS-1502, Trastuzumab mafodotin, Trastuzumab rezetecan, Vibostolimab, Vobramitamab duocarmazine, Zilovertamab vedotin, Suvemcitug, APX003, BD0801, TK001, sevacizumab, CTX-009, ES104, TR009, NOV1501, ABL001, DP303c, IAH0968, Kintuximab, gentuximab, cintuximab, MIL62, MRG002, MRG003, PM8002, MEDI5752, MCLA-128, AGEN1181, L19-IL2, L19-TNF, IMC-F106C, JMT101, BL-B01D1, BAT1308, AZD2936, JS004, TAB004, FG-M108, M108, 9MW2821, BMS-986349, CC-93269, EM801, ABBV-181, ABBV-151, ARGX-115, DS-7300a, IBI343, SHR-A1921, or ASKB589. In some embodiments, the modified peptide epitope or mimotope comprises or consists of an amino acid sequence having at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mutations (substitutions, insertions, and / or deletions) compared to an unmodified peptide epitope or mimotope of one of these antibodies but is capable of being bound by that antibody.

[0343] In some embodiments, the modified epitope or mimotope is capable of being bound by an anti-DLL3 monoclonal antibody. In some embodiments, the modified epitope or mimotope comprises 1, 2, 3, 4, or 5, mutations compared to the epitope or mimotope of the anti-DLL3 monoclonal antibody. In some embodiments, the anti-DLL3 monoclonal antibody is tarlatamab. In some embodiments, the unmodified epitope or mimotope comprises or consists of SEQ ID NO: 439.III. Recombinant Polypeptide

[0344] In one aspect, the disclosure provides recombinant polypeptide comprising or consisting of the epitope or mimotope of the disclosure. In some embodiments, the epitope or mimotope is comprised within a tag. That is, in some embodimetns, the disclosure provides recombinant polypeptides comprising a tag, wherein the tag comprises or consists of the peptide epitope or the mimotope of the disclosure.

[0345] In some embodimetns, the epitope or mimotope is a modified epitope or mimotope of the disclosure.

[0346] In some embodiments, the peptide epitope or mimotope is expressed on the surface of a cell. In some embodiments, the recombinant polypeptide comprises an extracellular region that comprises the peptide epitope or mimotope. In some embodiments, the peptide epitope or mimotope within the recombinant polypeptide is capable of being bound by the corresponding binder (e.g., an antibody) when expressed on the surface of an engineered cell.

[0347] In some embodiments, the epitope or mimotope is derived from the group consisting of CCR4, CCR7, DLL3, TROP2, GUCY2C, CD20, RSV protein F, EGFR, PD-1, CD34, and CD52, or is capable of being bound by an antigen-binding molecule (e.g., antibody) that binds to one of these proteins. In some embodiments, the recombinant polypeptide comprises or consists of CCR4, CCR7, DLL3, TROP2, GUCY2C, CD20, RSV protein F, EGFR, PD-1, CD34, or CD52, or a functional fragment thereof that comprises the epitope or mimotope. In some embodiments, the epitope or mimotope is derived from CCR4, CCR7, DLL3, TROP2, GUCY2C, CD20, RSV protein F, EGFR, PD-1, CD34, or CD52 but incorporated into a different polypeptide to form the recombinant polypeptide; that is, other than the epitope or mimotope, the recombinant polypeptide is not derived from that protein. In some embodiments, the recombinant polypeptide comprises an epitope ormimotope that is derived from capable of being bound by an anti-CCR4 antibody but is otherwise not derived from CCR4.

[0348] In some embodiments, the epitope or mimotope within the recombinant polypeptide does not comprise a cysteine at its N-terminus. In some embodiments, the recombinant polypeptide does not comprise a cysteine immediately before the N-terminus of the modified peptide epitope or mimotope.

[0349] In some embodiments, the epitope or mimotope within the recombinant polypeptide does not comprise a cysteine at its C-terminus. In some embodiments, the recombinant polypeptide does not comprise a cysteine immediately after the C-terminus of the modified peptide epitope or mimotope.

[0350] In some embodiments, the recombinant polypeptide comprises one or more additional peptide epitope(s) or mimotope(s). In some embodimetns, the one or more additional peptide epitope(s) or mimotope(s) are the same as the modified peptide epitope. In some embodimetns, the one or more additional peptide epitope(s) or mimotope(s) are different from the modified peptide epitope. In some embodiments, the recombinant polypeptide comprises 1, 2, 3, or 4 additional peptide epitopes or mimotopes.

[0351] In some embodiments, the recombinant polypeptide comprises a binding region for an antigen. In some embodiments, the antigen is expressed on the surface of a target cell. In some embodiments, the antigen is an autoimmune-associated antigen. In some embodiments, the antigen is a cancer-associated antigen.

[0352] In some embodiments, the antigen for the binding region is selected from the group consisting of CD1, CDla, CDlb, CDlc, CDld, CDle, CD2, CD3d, CD3e, CD3g, CD3s, CD4, CD5, CD7, CD8a, CD8b, CD19, CD20, CD21, CD22, CD23, CD24, CD25, CD27, CD28, CD30, CD33, CD34, CD38, CD40, CD44v6, CD45, CD46, CD47 CD48, CD52, CD59, CD66, CD70, CD71, CD72, CD73, CD79A, CD79B, CD80 (B7.1), CD86 (B7.2), CD94, CD95, CD97, CD123, CD134, CD140 (PDGFR4), CD152, CD154, CD158, CD171, CD178, CD179, CD179a, CD181 (CXCR1), CD182 (CXCR2), CD183 (CXCR3), CD210, CD213A2, CD246, CD252, CD253, CD261, CD262, CD272, CD273 (PD-L2), CD274 (PD- Ll), CD276 (B7H3), CD279, CD295, CD339 (JAG1), CD340 (HER2), CDH17, CEA, CLECL1, CLL-1, CLDN6, CLDN18.2, CS1, DLL3, LY6G6D, GCC, p53R175H, PRAME, EGFR, EGFRvIII, FGFR2, AFP, CA125, MUC-1, MAGE, ALPI, alkaline phosphatase placental-like 2 (ALPPL2), B-cell maturation antigen (BCMA), green fluorescent protein (GFP), enhanced green fluorescent protein (eGFP), KLK2, KLK3, Mesothelin, IL13Ra2,signal regulatory protein a (SIRPa), TCRalpha, TCRbeta, TSHR, GD2, GD3, Tn Ag, cMET, Axl, R0R1, ROR2, GPC1, GPC2, GPC3, FLT3, TAG72, CEA, EPCAM, KIT (CD117), IL- 13Ra2, IL-l lRa, PSCA, PRSS21, VEGFR2, LewisY, PDGFRp, SSEA-4, folate receptor alpha, ERBB2 (Her2 / neu), MUC1, MUC16, NCAM, prostase, PAP, ELF2M, Ephrin B2, IGF -I receptor, CAIX, LMP2, gplOO, bcr-abl, tyrosinase, EphA2, STEAP1, STEAP2, fucosyl GM1, sLe, GM3, TGS5, HMWMAA, o-acetyl-GD2, folate receptor beta, TEM1 / CD248, TEM7R, ROPN1, GPRC5D, GPA33, CXORF61, ALK, Polysialic acid, PLAC1, GloboH, NY-BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP, WT1, NY-ESO-1, LAGE-la, MAGE-A1, legumain, HPV E6,E7, MAGE Al, ETV6- AML, sperm protein 17, XAGE1, Tie 2, MAD-CT-1, MAD-CT-2, Fos-related antigen 1, p53, p53 mutant, p53R175H, KRAS, mutant KRAS, KRAS G12D, prostein, surviving, telomerase, PCTA-l / Galectin 8, MelanA / MARTl, Ras mutant, hTERT, sarcoma translocation breakpoints, ML-IAP, ERG (TMPRSS2 ETS fusion gene), NA17, PAX3, androgen receptor, cyclin Bl, MYCN, RhoC, TRP-2, CYP1B1, BORIS, SART3, PAX5, OY-TES1, LCK, AKAP-4, SSX2, RAGE-1, human telomerase reverse transcriptase, RU1, RU2, intestinal carboxyl esterase, mut hsp70-2,LAIRl, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, FCRL5, IGLL1, PSMA, TROP2, citrullinated vimentin, the extracellular portion of the APRIL protein, and any combinations thereof.

[0353] In some embodiments, the cancer-associated antigen for the binding region is a cancer-ssociated antigen for a hematologic cancer. In some embodiments, the cancer is selected from B cell lymphoma, T cell lymphoma, chronic lymphocytic leukemia, and acute myeloid leukemia.

[0354] In some embodiments, the cancer-associated antigen for the binding region is a cancer-associated antigen for a solid tumor. In some embodiments, the cancer is a solid tumor selected from the group consisting of small cell lung cancer, prostate cancer, colorectal cancer, head and neck squamous cell carcinoma, breast cancer, melanoma, ovarian cancer, lung cancer, pancreatic cancer, stomach cancer, cervical cancer, hepatocellular carcinoma, glioma cancer, neuroblastoma cancer, and renal cell carcinoma. In some embodiments, the cancer-associated antigen for the solid tumor comprises or consists of KIR2D, NRP1, CD221, VEGFR, HGF, integrin a5 1, CD51, CD27, B7H3, NYESO1, EGFR, cMET, BRD4, EpCAM, SSX, Fibronectin EDB, integrin al ipi, DLL3, CD2, SSTR2, TROP2, KLK2, KLK3, STEAP2, STEAP1, PSMA, PSCA, LY6G6D, IL-la, CEA, C242 antigen, CanAg (MUC1 glycoform), CTAA16.88, UCY2C, IGF2BP3, p53R175H,NKG2D ligands, PRAME, MAGEA4, vimentin, HER2 / neu, CCR4, CCR5, R0PN1, ROR1, KK-LC-1, CA-125, folate receptor 1, folate receptor alpha, MUC1, MUC16, MSLN, CLDN6, WT1, ALPPL2, KRASG12D, KRASG12V, CD70, MSLN, CLDN18.2, HPV E6 / E7, GPC3, IL-13Ra, EGFRv3, EGFR806, GD2, or CA9. In some embodiments, the cancer-associated antigen for the solid tumor comprises or consists of KIR2D, NRP1, CD221, VEGFR, HGF, integrin a5bl, CD51, CD27, B7H3, NYES01, EGFR, cMET, BRD4, EpCAM, SSX, Fibronectin EDB, or Integrin al ipi. In some embodiments, the cancer-associated antigen for the solid tumor comprises or consists of KLK2, Ly6G6D, CLDN6, DLL3 or CD 19.

[0355] In some embodiments, the cancer-associated antigen for the binding region is selected from one listed in Table 2 supra.

[0356] In some embodiments, the antigen for the binding region is not CD 19. In some embodiments, the antigen for the binding region is not CCR4. In some embodiments, the antigen is not CCR7.

[0357] In some embodiments, the antigen for the binding region is Delta-like protein 3 (DLL3).

[0358] In some embodiments, the antigen for the binding region is a cancer-associated antigen of adult T-cell lymphoma (ATL) and / or cutaneous T-cell lymphoma (CTCL). In some embodiments, In some embodiments, the antigen for the binding region is a cancer- associated antigen of a Th2 cell, a cutaneous lymphocyte antigen-positive skin-homing T cell, and / or a Treg cell. In some embodiments, the antigen for the binding region is C-C chemokine receptor type 4 (CCR4).

[0359] In some embodiments, the binding region is derived from an antibody. For example, the binding region may comprise or consist of an scFv comprising the VH and / or VL derived from the antibody. In some embodiments, the antibody is [fam-]trastuzumab deruxtecan, (fam-trastuzumab deruxtecan-nxki), Adebrelimab, Ado-trastuzumab emtansine, Alemtuzumab, Amivantamab, Atezolizumab, Avelumab, Belantamab mafodotin (belantamab mafodotin-blmf), Bevacizumab, Blinatumomab, Brentuximab vedotin, Cadonilimab, Camrelizumab, CAP- 100, Catumaxomab, Cemiplimab (cemiplimab-rwlc), Cetuximab, Cetuximab saratolacan, Cosibelimab, Daratumumab, Dinutuximab, Disitamab vedotin, Dostarlimab, Durvalumab, Edrecolomab, Elotuzumab, Elranatamab, Emapalumab (emapalumab-lzsg), Enfortumab vedotin (enfortumab vedotin-ejfv), Enlonstobart, Envafolimab, Epcoritamab, Gemtuzumab ozogamicin, Glofitamab, Ibritumomab tiuxetan,Inotuzumab ozogamicin, Ipilimumab, Isatuximab (isatuximab-irfc), Loncastuximab tesirine, Margetuximab-cmkb, Mirvetuximab soravtansine, Mogamulizumab (mogamulizumab- kpkc), Mosunetuzumab, Moxetumomab pasudotox (moxetumomab pasudotox-tdfk), Naxitamab-gqgk, Necitumumab, Nimotuzumab, Nivolumab, Cipterbin, Obinutuzumab, Ofatumumab, Olaratumab, Panitumumab, Pembrolizumab, Penpulimab, Pertuzumab, Polatuzumab vedotin (polatuzumab vedotin-piiq), Prolgolimab, Pucotenlimab, Racotumomab, Ramucirumab, Relatlimab, Retifanlimab, Ripertamab, Rituximab, Sacituzumab govitecan (sacituzumab govitecan-hziy), Serplulimab, Sintilimab, Socazolimab, Sugemalimab, Tafasitamab (tafasitamab-cxix), Tagitanlimab, Talquetamab (talquetamab-tgvs), Tebentafusp, Teclistamab, Tislelizumab, Tisotumab vedotin, tisotumab vedotin-tftv, Toripalimab, Tositumomab-1131, Trastuzumab, Trastuzumab duocarmazine, Tremelimumab, Zimberelimab, Zolbetuximab, Zuberitamab, Odronextamab, Ivonescimab, Benmel Stobart, Trastuzumab botidotin, Iparomlimab, tuvonralimab, Tarlatamab, Patritumab deruxtecan, Sacituzumab tirumotecan, Zanidatamab, Linvoseltamab, Datopotamab deruxtecan, Anbenitamab, Anvatabart opadotin, Apamistamab-Iodine (1311), Bemarituzumab, Cetrelimab, Cobolimab, Domvanalimab, Emactuzumab, Erfonrilimab, Favezelimab, Felzartamab, Fianlimab, Finotonlimab, ABBV-383, Geptanolimab, Gotistobart, Ivuxolimab, Izalontamab, Lemzoparlimab, Luveltamab tazevibulin, Magrolimab, Mecbotamab vedotin, Monalizumab, Nofazinlimab, Nurulimab, Ociperlimab, Oleclumab, Onfekafusp alfa, Oregovomab, Ozekibart, Pivekimab sunirine, Quavonlimab, Retlirafusp alfa, Rosopatamab (177Lu-DOTA), Rulonilimab, Sasanlimab, Telisotuzumab vedotin, Tiragolumab, IKS014, LCB14-0110, FS-1502, Trastuzumab mafodotin, Trastuzumab rezetecan, Vibostolimab, Vobramitamab duocarmazine, Zilovertamab vedotin, Suvemcitug, APX003, BD0801, TK001, sevacizumab, CTX-009, ESI 04, TR009, NOV1501, ABL001, DP303c, IAH0968, Kintuximab, gentuximab, cintuximab, MIL62, MRG002, MRG003, PM8002, MEDI5752, MCLA-128, AGEN1181, L19-IL2, L19-TNF, IMC-F106C, JMT101, BL-B01D1, BAT1308, AZD2936, JS004, TAB004, FG-M108, M108, 9MW2821, BMS-986349, CC-93269, EM801, ABBV-181, ABBV-151, ARGX-115, DS-7300a, IBI343, SHR-A1921, or ASKB589.

[0360] In some embodiments, the recombinant polypeptide (e.g., CAR or TCR) comprises the peptide epitope or mimotope that is capable of being bound by a monoclonal antibody that binds to the same antigen (e.g., an autoimmune associated antigen or cancer associated antigen of the disclosure) as the binding region. For example, in some embodiments, thebinding region is for binding to DLL3 antigen, and the recombinant polypeptide comprises an epitope or mimotope that is capable of being bound by an anti-DLL3 monoclonal antibody.

[0361] In some embodiments, the peptide epitope or mimotope is separated from the binding region by no more than 10, no more than 15, no more than 20, no more than 25, no more than 30, no more than 35, no more than 40, no more than 45, or no more than 50 amino acids.

[0362] In some embodiments, the binding region is a single-domain antibody fragment, single-chain variable fragment, antibody variable fragment, complementary determining region 3 fragment, constrained FR3-CDR3-FR4 polypeptide, Fd fragment, antigen-binding fragment, fibronectin-derived 10th fibronectin type III domain, tenascin type III domain, ankyrin repeat motif domain, low-density -lipoprotein-receptor-derived A-domain, lipocalin, Kunitz domain, Protein- A-derived Z domain, gamma-B crystallin-derived domain, ubiquitin-derived domain, Sac7d-derived polypeptide, Fyn-derived SH2 domain, miniprotein, C-type lectin-like domain scaffold, a heavy-chain antibody domain derived from a camelid VHH fragment, heavy-chain antibody domain derived from cartilaginous fish, immunoglobulin new antigen receptor (IgNAR), VNAR fragment, diabody, triabody, tetrabody, bivalent minibody, bispecific tandem scFv, bispecific tandem VHH, or bispecific minibody.

[0363] In some embodiments, the peptide epitope or mimotope is located N-terminal to the binding region in the recombinant polypeptide. In some embodiments, the peptide epitope or mimotope is separated from the N-terminus of the binding region by no more than 40, no more than 35, no more than 30, no more than 25, no more than 20, no more than 15, no more than 10, no more than 5, no more than 4, no more than 3, no more than 2, no more than 1, or zero amino acid(s).

[0364] In some embodiments, the recombinant polypeptide comprises no more than 10, no more than 15, no more than 20, no more than 25, no more than 30, no more than 35, no more than 40, no more than 45, or no more than 50 amino acids N-terminal to the binding region(s), excluding the signal peptide. In some embodiments, the recombinant polypeptide comprises or consists of about 10 to about 15, about 15 to about 20, about 20 to about 25, about 25 to about 30, about 30 to about 35, about 35 to about 40, about 40 to about 45, or about 45 to about 50, amino acids N-terminal to the binding region(s), excluding the signal peptide.

[0365] In some embodiments, the peptide epitope or mimotope is located C-terminal to the binding region in the recombinant polypeptide. In some embodiments, the peptide epitope or mimotope is separated from the C-terminus of the binding region by zero, no more than 5, no more than 10, no more than 15, no more than 20, no more than 25, no more than 30, no more than 35, no more than 40, no more than 45, no more than 50, no more than 55, or no more than 60 amino acids.

[0366] In some embodiments, the recombinant polypeptide comprises no more than 10, no more than 15, no more than 20, the recombinant polypeptide comprises no more than 10, no more than 15, no more than 20, no more than 25, no more than 30, no more than 35, no more than 40, no more than 45, no more than 50, no more than 55, no more than 60, no more than 65, or no more than 70 amino acids after the C-terminus of the binding region.

[0367] In some embodiments, the peptide epitope or mimotope is located within the binding region.

[0368] In some embodiments, the binding region comprises a heavy chain variable domain (VH) and a light chain variable domain (VL). In some embodiments, the binding region comprises a single chain variable fragment (scFv) comprising a VH and a VL. In some embodiments, the VH and the VL is separated by a linker. In some embodiments, the peptide epitope or mimotope is located within the linker. In some embodiments, the linker comprises no more than 10, no more than 15, no more than 20, no more than 25, no more than 30, no more than 35, no more than 40, no more than 45, or no more than 50 amino acids. In some embodiments, the linker comprises or consists of about 10 to about 15, about 15 to about 20, about 20 to about 25, about 25 to about 30, about 30 to about 35, about 35 to about 40, about 40 to about 45, or about 45 to about 50, amino acids.

[0369] In some embodiments, the binding region comprises one or more variable heavy domains of heavy chain (VHHs). In some embodiments, the binding region comprises at least two VHHs separated by a linker. In some embodiments, the peptide epitope or mimotope is located within the linker. In some embodiments, the linker comprises no more than 10, no more than 15, no more than 20, no more than 25, no more than 30, no more than 35, no more than 40, no more than 45, or no more than 50 amino acids. In some embodiments, the linker comprises or consists of about 10 to about 15, about 15 to about 20, about 20 to about 25, about 25 to about 30, about 30 to about 35, about 35 to about 40, about 40 to about 45, or about 45 to about 50, amino acids.

[0370] In some embodiments, the binding region comprises a first binding domain and a second binding domain, wherein the first binding domain and the second binding domain are separated by a linker. In some embodiments, the peptide epitope or mimotope is located within the linker. In some embodiments, the linker comprises no more than 10, no more than 15, no more than 20, no more than 25, no more than 30, no more than 35, no more than 40, no more than 45, or no more than 50 amino acids. In some embodiments, the linker comprises or consists of about 10 to about 15, about 15 to about 20, about 20 to about 25, about 25 to about 30, about 30 to about 35, about 35 to about 40, about 40 to about 45, or about 45 to about 50, amino acids.

[0371] In some embodiments, the recombinant polypeptide comprises a second binding region located N-terminal or C-terminal to the binding region; optionally, wherein the binding region and the second binding region are separated by a linker. In some embodiments, the peptide epitope or mimotope is located within the linker. In some embodiments, the linker comprises no more than 10, no more than 15, no more than 20, no more than 25, no more than 30, no more than 35, no more than 40, no more than 45, or no more than 50 amino acids. In some embodiments, the linker comprises or consists of about 10 to about 15, about 15 to about 20, about 20 to about 25, about 25 to about 30, about 30 to about 35, about 35 to about 40, about 40 to about 45, or about 45 to about 50, amino acids.

[0372] In some embodiments, the peptide epitope or mimotope is located in between the binding region(s) and a transmembrane region. In some embodiments, the peptide epitope or mimotope is located within a hinge region that connects the binding region(s) to a transmembrane region. In some embodiments, the hinge region comprises no more than 10, no more than 15, no more than 20, no more than 25, no more than 30, no more than 35, no more than 40, no more than 45, no more than 50, no more than 55, no more than 60, no more than 65, or no more than 70 amino acids. In some embodiments, the hinge region comprises or consists of about 10 to about 15, about 15 to about 20, about 20 to about 25, about 25 to about 30, about 30 to about 35, about 35 to about 40, about 40 to about 45, about 45 to about 50, about 50 to about 55, about 55 to about 60, about 60 to about 65, or about 65 to about 70, amino acids.

[0373] In some embodiments, the extracellular region of the recombinant polypeptide (e.g., CAR or TCR) comprises no more than 200, no more than 250, no more than 300, no more than 350, no more than 400, no more than 450, or no more than 500, amino acids.

[0374] In some embodiments, the extracellular region of the recombinant polypeptide (e.g., CAR or TCR) comprises about 200 to about 250, about 250 to about 300, about 300 to about 350, about 350 to about 400, about 400 to about 450, or about 450 to about 500, amino acids. In some embodiments, the extracellular region of the recombinant polypeptide (e.g., CAR or TCR) comprises no more than 400 amino acids.

[0375] In some embodiments, the recombinant polypeptide (e.g., CAR or TCR) comprises no more than 200, no more than 250, no more than 300, no more than 350, no more than 400, no more than 450, no more than 500, no more than 550, no more than 600, no more than 650, no more than 700, no more than 750, or no more than 800, amino acids. In some embodiments, the recombinant polypeptide (e.g., CAR or TCR) comprises about 200 to about 250, about 250 to about 300, about 300 to about 350, about 350 to about 400, about 400 to about 450, about 450 to about 500, about 500 to about 550, about 550 to about 600, about 600 to about 650, about 650 to about 700, about 700 to about 750, or about 750 to about 800, amino acids. In some embodiments, the recombinant polypeptide (e.g., CAR or TCR) comprises no more than 600 amino acids. In some embodiments, the recombinant polypeptide (e.g., CAR or TCR) comprises no more than 550 amino acids.

[0376] In some embodiments, the recombinant polypeptide comprises or consists of an engineered receptor.

[0377] In some embodiments, the recombinant polypeptide does not comprise a chimeric antigen receptor (CAR) or a recombinant T cell receptor (TCR).

[0378] In some embodiments, the recombinant polypeptide comprises or consists of a T cell receptor fusion construct (TRuC). In some embodiments, the TRuC is derived from a CD3 protein and comprises a binding region. In some embodiments, the CD3 protein is CD3s.

[0379] In some embodiments, the recombinant polypeptide comprises or consists of a chimeric antigen receptor (CAR) or a recombinant T cell receptor (TCR).

[0380] In some embodiments, the recombinant polypeptide comprises or consists of a receptor comprising: (a) a heavy chain variable region (VH) located before the N-terminus of a constant region of TCRalpha, and a light chain variable region (VL) located before the N-terminus of a constant region of TCRbeta; or (b) a heavy chain variable region (VH) located before the N-terminus of a constant region of TCRbeta, and a light chain variable region (VL) located before the N-terminus of a constant region of TCRalpha. In some embodiments, , at least one of the constant region of TCRalpha or the constant region ofTCRbeta are an endogenous constant region of TCRalpha or an endogenous constant region of TCRbeta. In some embodiments, the receptor is a T-body.

[0381] In some embodiments, the peptide epitope or mimotope is located between the VH and the constant region of TCRalpha. In some embodiments, the peptide epitope or mimotope is located between the VL and the constant region of TCRalpha. In some embodiments, the peptide epitope or mimotope is located between the VH and the constant region of TCRbeta. In some embodiments, the peptide epitope or mimotope is located between the VL and the constant region of TCRbeta. In some embodiments, the peptide epitope or mimotope is located N-terminal to the VH. In some embodiments, the peptide epitope or mimotope is located N-terminal to the VL. In some embodiments, the peptide epitope or mimotope is located C-terminal to the constant region of TCRalpha. In some embodiments, the peptide epitope or mimotope is located C-terminal to the constant region of TCRbeta.

[0382] In some embodiments, the recombinant polypeptide comprises an extracellular portion comprising the binding region and the peptide epitope or mimotope.

[0383] In some embodiments, the recombinant polypeptide comprises a transmembrane region. In some embodiments, the recombinant polypeptide (e.g., CAR or TCR) comprises, from N-terminus to C-terminus, the binding region(s), a hinge region, a transmembrane region. In some embodiments, the transmembrane region comprises a polypeptide sequence at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 421. In some embodiments, the hinge region comprises a polypeptide sequence at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 420.

[0384] In some embodiments, the recombinant polypeptide (e.g., CAR or TCR) further comprises an intracellular region. In some embodiments, the intracellular region comprises one or more intracellular signaling domains. In some embodiments, the intracellular domain is located C-terminal to the transmembrane region.

[0385] In some embodiments, the one or more intracellular signaling domains are derived from a protein selected from the group consisting of CD3zeta, lxxCD3zeta, CD28, 4- IBB, 0X40, IL2Rb turbodomains, MYD88 / CD40, MYD88 / CD40 inducible costimulatory domain, CD2, B7-1 / CD80; B7-2 / CD86; B7-H1 / PD-L1; B7-H2; B7-H3; B7-H4; B7-H6; B7- H7; BTLA / CD272; CD28; CTLA-4; Gi24 / VISTA / B7-H5; ICOS / CD278; PD-1; PD-L2 / B7- DC; PDCD6); 4-1BB / TNFSF9 / CD137; 4-1BB Ligand / TNFSF9; BAFF / BLyS / TNFSF13B;BAFF R / TNFRSF13C; CD27 / TNFRSF7; CD27 Ligand / TNFSF7; CD30 / TNFRSF8; CD30 Ligand / TNFSF8; CD40 / TNFRSF5; CD40 / TNFSF5; CD40 Ligand / TNFSF5;DR3 / TNFRSF25; GITR / TNFRSF18; GITR Ligand / TNFSF18; HVEM / TNFRSF14; LIGHT / TNFSF14; Lymphotoxin-alpha / TNF-beta; OX40 / TNFRSF4; 0X40 Ligand / TNFSF4; RELT / TNFRSF19L; TACI / TNFRSF13B; TL1A / TNFSF15; TNF-alpha; TNF RII / TNFRSF1B); 2B4 / CD244 / SLAMF4; BLAME / SLAMF8; CD2; CD2F-10 / SLAMF9; CD48 / SLAMF2; CD58 / LFA-3; CD84 / SLAMF5; CD229 / SLAMF3; CRACC / SLAMF7; NTB- A / SLAMF6; SLAM / CD150); CD2; CD7; CD53; CD82 / Kai-1; CD90 / Thyl; CD96; CD160; CD200; CD300a / LMIRl; HLA Class I; HLA-DR; Ikaros; Integrin alpha 4 / CD49d; Integrin alpha 4 beta 1; Integrin alpha 4 beta 7 / LPAM-l; LAG-3; TCL1A; TCL1B; CRTAM; DAP12; Dectin- 1 / CLEC7A; DPPIV / CD26; EphB6; TIM- 1 / KIM- 1 / HA VCR; TIM-4; TSLP; TSLP R; lymphocyte function associated antigen-1 (LFA-1); NKG2C, an immunoreceptor tyrosine based activation motif (ITAM), CD27, CD134 / OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD7, LIGHT, NKG2C, and any combinations thereof.

[0386] In some embodiments, the one or more intracellular signaling domains comprises singaling domain(s) derived from 4- IBB and / or CD3zeta. In some embodiments, the intracellular region comprises the signaling domain derived from 4- IBB and the signaling domain derived from CD3zeta. In some embodiments, the intracellular region comprises, from N-term to C-term, (i) the signaling domain derived from 4- IBB and (ii) the signaling domain derived from CD3zeta. In some embodiments, the signaling domain derived from 4- 1BB comprises a polypeptide sequence at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 422. In some embodiments, the signaling domain derived from CD3zeta comprises a polypeptide sequence at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 423. In some embodiments, the signaling domain derived from CD3zeta comprises a polypeptide sequence at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 488, wherein the amino acid corresponding to position 14 of SEQ ID NO: 488 is lysine.

[0387] In some embodiments, the recombinant polypeptide comprises a glycosylphosphatidylinositol (GPI) anchor.

[0388] In some embodiments, the TCR is TCRalpha. In some embodiments, the TCR is TCRbeta. In some embodiments, the binding region of the TCR comprises a variable region and a constant region. In some embodiments, the peptide epitope or mimotope is located between the variable region and the constant region. In some embodiments, the variable region and the constant region in the TCR are separated by no more than 10, no more than 15, no more than 20, no more than 25, no more than 30, no more than 35, no more than 40, no more than 45, or no more than 50 amino acids. In some embodiments, the variable region and the constant region in the TCR are separated by about 10 to about 15, about 15 to about 20, about 20 to about 25, about 25 to about 30, about 30 to about 35, about 35 to about 40, about 40 to about 45, or about 45 to about 50, amino acids. In some embodiments, the peptide epitope or mimotope is located N-terminal to the variable region of the TCR. In some embodiments, the peptide epitope or mimotope is located C-terminal to the constant region of the TCR.

[0389] In some embodiments, an immune cell expressing the recombinant polypeptide (e.g., receptor) of the disclosure exhibits one or more of the following: (a) improved binding to the antigen expressed on the surface of the target cell; (b) reduced recombinant polypeptide mediated antigen-independent signaling and / or activation of the immune cell; and / or (c) reduced cross-linking between the recombinant polypeptide expressed on the cell surface of the immune cell, compared to a control immune cell expressing a control recombinant polypeptide that does not comprise the epitope or mimotope.

[0390] In some embodiments, an immune cell expressing the recombinant polypeptide (e.g., receptor) of the disclosure exhibits one or more of the following: (a) comparable level of binding to the antigen expressed on the surface of the target cell; (b) comparable level of recombinant polypeptide mediated antigen-independent signaling and / or activation of the immune cell; and / or (c) comparable level of cross-linking between the recombinant polypeptide expressed on the cell surface of the immune cell, compared to a control immune cell expressing a control recombinant polypeptide that does not comprise the epitope or mimotope.In some embodiments, the recombinant polypeptide comprises a modified epitope or mimotope of the disclosure. In some embodiments, an immune cell expressing the recombinant polypeptide (e.g., receptor) comprising the modified epitope or mimotope exhibits one or more of the following: (a) improved binding to the antigen expressed on the surface of the target cell; (b) reduced recombinant polypeptide mediated antigen-independent signaling and / or activation of the immune cell; and / or (c) reduced cross-linking between the recombinant polypeptide expressed on the cell surface of the immune cell, compared to a control immune cell expressing a control recombinant polypeptide that comprise the corresponding unmodified epitope or mimotope. In some embodiments, an immune cell expressing the recombinant polypeptide (e.g., receptor) comprising the modified epitope or mimotope exhibits one or more of the following: (a) comparable level of binding to the antigen expressed on the surface of the target cell; (b) comparable level of recombinant polypeptide mediated antigen-independent signaling and / or activation of the immune cell; and / or (c) comparable level of cross-linking between the recombinant polypeptide expressed on the cell surface of the immune cell, compared to a control immune cell expressing a control recombinant polypeptide that comprises the corresponding unmodified epitope or mimotope.IV. Engineered Receptors and Components Thereof

[0391] In one aspect, the disclosure provides engineered receptors, such as Chimeric Antigen Receptors (CARs), T Cell Receptors (TCRs), T cell receptor fusion constructs (TRuCs), and other synthetic Receptors. In some embodiments, the engineered receptor is expressed in a cell. In some embodiments, the engineered receptor comprises the epitope or mimotope of the disclosure. In some embodiments, the engineered receptor is co-expressed with a separate recombinant polypeptide comprising the epitope or mimotope.

[0392] In some embodiments, the engineered receptor is selected form a chimeric antigen receptor (CAR), a T cell receptor (TCR) such as a recombinant TCR, a T cell receptor fusion construct (TRuC), a chimeric auto antigen receptor (CAAR), a B-cell autoantibody receptor (BAR), a chimeric cytokine receptor (CCR), a chimeric chemokine receptor, a synthetic RIP family receptor, a T-body, a synthetic notch receptor (synNotch), a synthetic intramembrane proteolysis receptor (SNIPR), a synthetic Robo receptor (SynRobo receptor), a synthetic RoboNotch receptor (SynRobo receptor), a Modular Extracellular Sensor Architecture (MESA) receptor, a Tango receptor, a ChaCha receptor, a generalized extracellular molecule sensor (GEMS) receptor, a growth factor receptor, a cytokine receptor, a chemokine receptor, a switch receptor, an adhesion molecule, an integrin, an inhibitory receptor, a stimulatory receptor, an immunoreceptor tyrosine-based activation motif (ITAM)-containing receptor, an immunoreceptor tyrosine-based inhibition motif (ITIM)-containing receptor, a hormone receptor, and a receptor tyrosine kinase.

[0393] In some embodiments, the engineered receptor comprises one or more domains that combine an antigen -binding domain (e.g., derived from an antibody or antibody fragment) that provides a capability for binding a desired antigen (e.g., tumor antigen) with intracellular signaling domains. In some embodiments, the intracellular signaling domain is an activating intracellular domain portion, such as a T cell activating domain, providing a primary activation signal. In some embodiments, the intracellular signaling domain contains or additionally contains a costimulatory signaling domain to facilitate effector functions. Upon binding to the molecule, e.g., antigen, the receptor generally delivers an immunostimulatory signal, such as an ITAM-transduced signal, into the cell, thereby promoting an immune response targeted to the disease or condition. In some embodiments, such receptors upon introduction into immune cells can modulate the cell activity, and, in some cases, can modulate cell differentiation or homeostasis, thereby resulting in genetically engineered cells with improved longevity, survival and / or persistence in vivo, such as for use in adoptive cell therapy methods.

[0394] In some embodiments, the engineered receptor is constructed with a capability for binding a particular antigen (or marker or ligand), such as an antigen expressed in a particular cell type to be targeted by adoptive therapy, e.g., a cancer marker, and / or an antigen intended to induce a dampening response, such as an antigen expressed on a normal or non-diseased cell type. Thus, the engineered receptor typically includes in its extracellular portion one or more antigen binding molecules, such as one or more antigen-binding fragment, domain, or portion, or one or more antibody-derived variable domains, and / or antibody-derived molecules.

[0395] In some embodiments, an engineered cell provided herein (e.g., primary or iPSC- derived T cell or primary or iPSC-derived NK cell) includes a recombinant nucleic acid encoding the engineered receptor, wherein the recombinant nucleic acid is inserted in a genomic locus. In some embodiments, the recombinant nucleic acid is inserted into a safe harbor locus, such as but not limited to, an AAVS1 locus, CCR5 locus, CLYBL locus, ROSA26 locus, SHS231 locus, F3 (also known as CD142) locus, MICA locus, MICB locus, LRP1 (also known as CD91) locus, HMGB1 locus, ABO locus, RHD locus, FUT1 locus, or KDM5D locus. In some embodiments, the recombinant nucleic acid is inserted in a B2M locus, CIITA locus, CARD9 locus, CARDIO locus, CARD 14 locus, TRAC locus, TRBC1 locus, TRBC2 locus, CARD11 locus, PIK3R3 locus, PD-1 locus, or CTLA-4 locus. Anysuitable method can be used to insert the recombinant nucleic acidinto the genomic locus of the cell including the gene editing methods described herein (e.g., a CRISPR / Cas system).

[0396] In accordance with the present disclosure, on-off or other types of control switch techniques may be incorporated herein. These techniques may comprise use of dimerization domains and optional activators of such domain dimerization, e.g., as disclosed by Wu et al., Science 2015 Oct; 350(6258):aab4077 utilizing FKBP / Rapalog dimerization systems in certain cells, the contents of which are incorporated by reference herein in their entirety.Additional dimerization technology is described in, e.g., Fegan et al. Chem. Rev. 2010, 110, 3315-3336 as well as U.S. Pat. Nos. 5,830,462; 5,834,266; 5,869,337; and 6,165,787, the contents of each of which is also incorporated by reference herein with respect to dimerization technology. Additional dimerization pairs may comprise cyclosporine- A / cyclophilin, receptor, estrogen / estrogen receptor (optionally using tamoxifen, 4- hydroxytamoxifen, or endoxifen), glucocorticoids / glucocorticoid receptor, tetracycline / tetracycline receptor, and / or vitamin D / vitamin D receptor. Further examples of dimerization technology may be found in e.g., WO 2014 / 127261, WO 2015 / 090229, US 2014 / 0286987, US 2015 / 0266973, US 2016 / 0046700, U.S. Pat. No. 8,486,693, US 2014 / 0171649, and US 2012 / 0130076, the contents of which are further incorporated by reference herein in their entirety. a. Chimeric Antigen Receptors

[0397] In some embodiments, the CAR is selected from the group consisting of a first generation CAR, a second generation CAR, a third generation CAR, and a fourth generation CAR. In some embodiments, the CAR is or comprises a first generation CAR comprising an antigen binding region, a transmembrane domain, and at least one signaling domain (e.g., one, two or three signaling domains). In some embodiments, the CAR is or comprises a second generation CAR comprising an antigen binding region, a transmembrane domain, and at least two signaling domains. In some embodiments, the CAR is or comprises a third generation CAR comprising an antigen binding region, a transmembrane domain, and at least three signaling domains. In some embodiments, the CAR is or comprises a fourth generation CAR comprising an antigen binding region, a transmembrane domain, three or four signaling domains, and a domain which upon successful signaling of the CAR induces expression of a cytokine gene.

[0398] In some embodiments, the CAR is or comprises a first generation CAR. In some embodiments, a first generation CAR comprises an antigen binding region, a transmembrane domain, and signaling domain. In some embodiments, a signaling domain mediates downstream signaling during T cell activation.

[0399] In some embodiments, the CAR is or comprises a second generation CAR. In some embodiments, a second generation CAR comprises an antigen binding region, a transmembrane domain, and two signaling domains. In some embodiments, a signaling domain mediates downstream signaling during T cell activation. In some embodiments, a signaling domain is a costimulatory domain. In some embodiments, a costimulatory domain enhances cytokine production, CAR T cell proliferation, and / or CAR T cell persistence during T cell activation.

[0400] In some embodiments, the CAR is or comprises a third generation CAR. In some embodiments, a third generation CAR comprises an antigen binding region, a transmembrane domain, and at least three signaling domains. In some embodiments, a signaling domain mediates downstream signaling during T cell activation. In some embodiments, a signaling domain is a costimulatory domain. In some embodiments, a costimulatory domain enhances cytokine production, CAR T cell proliferation, and or CAR T cell persistence during T cell activation. In some embodiments, a third generation CAR comprises at least two costimulatory domains. In some embodiments, the at least two costimulatory domains are not the same.

[0401] In some embodiments, the CAR is or comprises a fourth generation CAR. In some embodiments, a fourth generation CAR comprises an antigen binding region, a transmembrane domain, and at least two, three, or four signaling domains. In some embodiments, a signaling domain mediates downstream signaling during T cell activation. In some embodiments, a signaling domain is a costimulatory domain. In some embodiments, a costimulatory domain enhances cytokine production, CAR T cell proliferation, and or CAR T cell persistence during T cell activation.

[0402] In some embodiments, the CAR comprises an antigen binding region and a transmembrane domain, but does not comprise any signaling domain. In some embodiments, the transmembrane domain of such CARs binds to another signaling protein. In some embodiments, such CARs comprise a domain that interacts with another signaling protein. In some embodiments, such CARs are capable of activating the signaling protein it interacts with in the presence of the antigen.

[0403] In some embodiments, the antigen binding region of the CAR is or comprises an antigen-binding fragment of an antibody. In some embodiments, the antibody is a human antibody or humanized antibody. In some embodiment, the antibody is a monoclonal antibody.

[0404] In some embodiments, the CAR includes an antigen-binding portion or portions of an antibody molecule, such as a single-chain antibody fragment (scFv) derived from the variable heavy (VH) and variable light (VL) chains of a monoclonal antibody (mAh), or a single domain antibody (sdAb), such as sdFv, nanobody, VHH and VNAR. In some embodiments, an antigen binding fragment comprises antibody-derived variable regions joined by a flexible linker.

[0405] In some embodiments, the antibody or antigen-binding fragment thereof is a single chain antibody fragment, such as a single chain variable fragment (scFv) or a diabody or a single domain antibody (sdAb). In some embodiments, the antibody or antigen-binding fragment is a single domain antibody comprising only the VH region. In some embodiments, the antibody or antigen binding fragment is an scFv comprising a heavy chain variable (VH) region and a light chain variable (VL) region.

[0406] In some embodiments, the CAR includes a binding portion or portions of an antibody molecule, such as a heavy chain variable (VH) region and / or light chain variable (VL) region of the antibody, e.g., an scFv antibody fragment. The chimeric receptors, such as CARs, generally include an extracellular antigen binding region, such as a portion of an antibody molecule, generally a variable heavy (VH) chain region and / or variable light (VL) chain region of the antibody, e.g., an scFv antibody fragment.

[0407] In some embodiments, the antibody or antigen binding fragment, contains a heavy and / or light chain variable (VH or VL) region sequence as described, or a sufficient antigenbinding portion thereof. In some embodiments, the antigen-binding fragment, contains a VH region sequence or sufficient antigen -binding portion thereof that contains a CDR-H1, CDR-H2 and / or CDR-H3 as described. In some embodiments, the antigen-binding fragment, contains a VL region sequence or sufficient antigen-binding portion that contains a CDR-L1, CDR-L2 and / or CDR-L3 as described. In some embodiments, the antigen-binding fragment, contains a VH region sequence that contains a CDR-H1, CDR-H2 and / or CDR-H3 as described and contains a VL region sequence that contains a CDR-L1, CDR-L2 and / or CDR-L3 as described. Also among the antibodies are those having sequences at least at orabout 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to such a sequence.

[0408] In some embodiments, the antibody is a single domain antibody (sdAb) comprising only a VH region sequence or a sufficient antigen-binding portion thereof, such as any of the above described VH sequences (e.g., a CDR-H1, a CDR-H2, a CDR-H3 and / or a CDR-H4).

[0409] In some embodiments, an antibody provided herein or antigen-binding fragment thereof comprising a VH 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 contains a VH region and a VL region, or a sufficient antigen binding portion of a VH and VL region. In such embodiments, a VH region sequence can be any of the above described VH sequence. In some such embodiments, the antibody is an antigen binding fragment, such as a Fab or an scFv. In some such embodiments, the antibody is a full-length antibody that also contains a constant region.

[0410] In some embodiments, the nucleotide sequence encoding a CAR may be derived from a mammalian sequence, for example, a mouse sequence, a primate sequence, a human sequence, or combinations thereof. In the cases where the nucleotide sequence encoding a CAR is non-human, the sequence of the CAR may be humanized. The nucleotide sequence encoding a CAR may also be codon-optimized for expression in a mammalian cell, for example, a human cell. In any of these embodiments, the nucleotide sequence encoding a CAR may be at least 80% identical e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to any of the nucleotide sequences disclosed herein. The sequence variations may be due to codon- optimalization, humanization, restriction enzymebased cloning scars, and / or additional amino acid residues linking the functional domains, etc.

[0411] In certain embodiments, the CAR can be re-engineered as a chimeric autoantibody receptor (CAAR) to selectively deplete autoreactive immune cells. In certain embodiments, CAARs are engineered to target autoantibodies present on immune cells. Exemplary target antigens for CAARs include, but are not limited to, DSG3 (associated with pemphigus volgaris); factor VIII (FVIII)(associated with haemophilia). In any of these embodiments, the extracellular binding domain of the CAR can be codon-optimized for expression in a host cell or have variant sequences to increase functions of the extracellular binding domain.

[0412] In some embodiments, the CAR is a bicistronic CAR. In some embodiments, the immune cell of the disclosure comprises a bicistronic CAR. Bicistronic CARs can comprisetwo CARs that bind different targets and are encoded by a single vector. A bicistronic CAR may comprise a first CAR sequence and a second CAR sequence expressed as a single polypeptide comprising a cleavable linker between the first and second CARs. Non-limiting examples of first and / or second CAR sequences include CD19,CD20, BCMA, CD22, CD70, DLL3, LY6G6D, Claudin 6, GCC, p53R175H, and PRAME. An exemplary cleavable linker is Furin-GSG-T2A (see, e.g., Chng et al. MAbs. 2015 Mar-Apr; 7(2): 403-412, which is herein incorporated by reference with respect to cleavable linkers; see also Guedan et al. Mol Ther Methods Clin Dev. 2019 Mar 15; 12: 145-156, which is incorporated herein by reference with respsect to bicistronic CAR design).

[0413] In some embodiments, the CAR is a bispecific CAR. In some embodiments, the immune cell of the disclosure comprises a bispecific CAR. In some embodiments, a first binding motif and a second binding motif (e.g., distinct anti-CD20 and anti-CD19 binding motifs) are both comprised in single bispecific CAR. In such bispecific CARs, a CAR molecule itself may be engineered to recognize more than one antigen. In tandem bispecific CARs, the first and second binding motifs are extracellular and may be characterized as a membrane-proximal binding motif and a membrane-distal binding motif.

[0414] In some embodiments, the CAR further comprises a domain which upon successful signaling of the CAR induces expression of a cytokine gene. In some embodiments, a cytokine gene is endogenous or exogenous to a target cell comprising a CAR which comprises a domain which upon successful signaling of the CAR induces expression of a cytokine gene. In some embodiments, a cytokine gene encodes a pro-inflammatory cytokine. In some embodiments, a cytokine gene encodes IL-1, IL-2, IL-9, IL- 12, IL- 18, TNF, or IFN- gamma, or functional fragment thereof. In some embodiments, a domain which upon successful signaling of the CAR induces expression of a cytokine gene is or comprises a transcription factor or functional domain or fragment thereof. In some embodiments, a domain which upon successful signaling of the CAR induces expression of a cytokine gene is or comprises a transcription factor or functional domain or fragment thereof. In some embodiments, a transcription factor or functional domain or fragment thereof is or comprises a nuclear factor of activated T cells (NF AT), an NF-kB, or functional domain or fragment thereof. See, e.g., Zhang. C. et al., Engineering CAR-T cells. Biomarker Research. 5:22 (2017); WO 2016126608; Sha, H. et al. Chimaeric antigen receptor T-cell therapy for tumour immunotherapy. Bioscience Reports Jan 27, 2017, 37 (1).

[0415] A skilled artisan is familiar with CARs and different components and configurations of CARs. Any known CAR can be employed in connection with the provided embodiments. In addition to the CARs described herein, various CARs and nucleotide sequences encoding the same are known in the art and would be suitable for engineering cells as described herein. See, e.g., W02013040557; W02012079000; W02016030414;Smith T, et al., Nature Nanotechnology. 2017. DOI: 10.1038 / NNAN0.2017.57, the disclosures of which are herein incorporated by reference. Exemplary features and components of a CAR are described in the following subsections.1. Antigen Binding Region

[0416] In some embodiments, a CAR antigen binding region is or comprises an antibody or antigen-binding portion thereof. In some embodiments, a CAR antigen binding region is or comprises an scFv or Fab.

[0417] In some embodiments, an antigen binding region binds to a cell surface antigen of a cell. In some embodiments, a cell surface antigen is characteristic of (e.g., expressed by) a particular or specific cell type. In some embodiments, a cell surface antigen is characteristic of more than one type of cell.

[0418] In some embodiments, the antigen may be an antigen that is expressed on tumor cells, or an antigen that is characteristic of an autoimmune or inflammatory disease. In some embodiments, the antigen may be an antigen that is exclusively or preferentially expressed on tumor cells, or an antigen that is characteristic of an autoimmune or inflammatory disease. In some embodiments, the antigen binding region targets an antigen characteristic of a neoplastic cell. For instance, the antigen binding region targets an antigen expressed by a neoplastic or cancer cell. In some embodiments, the antigen binding region binds a tumor associated antigen. In some embodiments, the antigen characteristic of a neoplastic cell (e.g., antigen associated with a neoplastic or cancer cell) or a tumor associated antigen is selected from a cell surface receptor, an ion channel-linked receptor, an enzyme-linked receptor, a G protein-coupled receptor, receptor tyrosine kinase, tyrosine kinase associated receptor, receptor-like tyrosine phosphatase, receptor serine / threonine kinase, receptor guanylyl cyclase, histidine kinase associated receptor.

[0419] In some embodiments, the target antigen is an antigen that includes, but is not limited to, ALPPL2, Epidermal Growth Factor Receptors (EGFR), EGFRvIII, ErbBl / EGFR, ErbB2 / HER2, ErbB3 / HER3, ErbB4 / HER4, Fibroblast Growth Factor Receptors (FGFR), FGFR1, FGFR2, FGFR3, FGFR4, FGFR5, FGFR6, FGFR7, FGFR18, FGFR21, VascularEndothelial Growth Factor Receptors (VEGFR), VEGFR1, VEGFR2, VEGR3, VEGF-A, VEGF-B, VEGF-C, VEGF-D, PIGF, RET Receptor, Eph Receptor Family, EphAl, EphA2, Eph A3, EphA4, EphA5, EphA6, EphA7, EphA8, EphA9, EphAlO, EphBl, EphB2. EphB3, EphB4, EphB6, ELF2M, CXCR1, CXCR2, CXCR3, CXCR4, CXCR6, CCR1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR8, CFTR, CIC-1, CIC-2, CIC-4, CIC-5, CIC-7, CIC-Ka, CIC-Kb, Bestrophins, TMEM16A, GAB A receptor, glycin receptor, ABC transporters, NAV1.1, NAVI.2, NAVI.3, NAVIA, NAVI.5, NAVIA, NAVI.7, NAVI.8, NAVI.9, sphingosine- 1 -phosphate receptor (S1P1R), NMDA channel, transmembrane protein, multispan transmembrane protein, T-cell receptor motifs; T-cell alpha chains; T-cell beta chains; T-cell y chains; T-cell 6 chains, GCC, CCR7, CD1, CDla, CDlb, CDlc, CDld, CDle, CD2, CD3d, CD3e, CD3g, CD3s, CD3, CD4, CD5, CD7, CD8a, CD8b, CD8, CDllb, CDl lc, CD16, CD19, CD20, CD21, CD22, CD23, CD24, CD25, CD27, CD28, CD30, CD33, CD34, CD35, CD38, CD40, CD45RA, CD45RO, CD46, CD47, CD48, CD52, CD56, CD59, CD62L, CD66, CD68, CD70, CD71, CD72, CD73, CD79A, CD79B, CD80 (B7.1), CD86 (B7.2), CD94, CD95, CD97, CD117, CD123, CD127, CD133, CD134, CD140 (PDGFR4), CD152, CD154, CD158, CD171, CD178, CD179, CD179a, CD181 (CXCR1), CD182 (CXCR2), CD183 (CXCR3), CD210, CD213A2, CD134, CD137 (4-1 BB), CD163, CD246, CD252, CD253, CD261, CD262, CD272, CD273 (PD-L2), CD274 (PD-L1), CD276 (B7H3), CD279, CD295, CD339 (JAG1), CD340 (HER2), CDH17, F4 / 80, IL-4Ra, Sca-1 , CTLA-4, GITR, GARP, LAP, granzyme B, LFA-1, transferrin receptor, NKp46, perforin, CD4+, Thl, Th2, Thl7, Th40, Th22, Th9, Tfh, Canonical Treg, FoxP3+, Tri, Th3, Tregl7, TREG, CDCP, NT5E, EpCAM, CEA, gpA33, Mucins, Carbonic anhydrase IX, Folate binding protein, Gangliosides (e.g., CD2, CD3, GM2), Lewis-y2, aVp3, a5pl, ErbBl / HER2, ErB3, c-MET, IGF1R, TRAIL-R1, TRAIL-R2, RANKL, FAP, Tenascin, PDL-1, BAFF, HD AC, ABL, FLT3, KIT, MET, RET, IL- Ip, ALK, RANKL, mTOR, CTLA-4, IL-6, IL-6R, JAK3, BRAF, PTCH, Smoothened, PIGF, ANPEP, TIMP1, PLAUR, PTPRJ, LTBR, or ANTXR1, Folate receptor alpha (Fra), ERBB2 (Her2 / neu), IL-13Ra2, Mesothelin, TSHR, CD171, CD 179a, CLECL1, CS-1, CLL-1, GD2, GD3, BCMA (CD269), MUC16 (CAI 25), LI CAM, LeY, MSLN, IL13Ral, Ll-CAM, Tn Ag, prostate specific membrane antigen (PSMA), R0R1, GM3, CD44v6, B7H3 (CD276), KIT, interleukin- 11 receptor a (IL-1 IRa), PSCA, PRSS21, LewisY, platelet-derived growth factor receptor-beta (PDGFR-beta), S SEA-4, DLL3, MUC1, NCAM, Prostase, PAP, Ephrin B2, IGF-1 receptor, CAIX, LMP2, gplOO, bcr-abl, tyrosinase, Fucosyl GM1, sLe, TGS5, HMWMAA, o-acetyl-GD2, Folate receptor beta, TEM1 / CD248, TEM7R, CLDN6, CLDN18.2, CX0RF61, CD97, CD 179a, ALK, Polysialic acid, PLAC1, GloboH, NY-BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP, WT1, NY-ESO- 1, LAGE-la, MAGE-A1, legumain, HPV E6, E7, MAGE-A4, ETV6-AML, sperm protein 17, XAGE1, Tie 2, MAD- CT-1, MAD-CT-2, Major histocompatibility complex class I-related gene protein (MR1), urokinase-type plasminogen activator receptor (uPAR), Fos-related antigen 1, p53, p53 mutant, prostein, survivin, telomerase, PCTA-l / Galectin 8, MelanA / MARTl, Ras mutant, hTERT, sarcoma translocation breakpoints, ML-IAP, ERG (TMPRSS2 ETS fusion gene), NA17, PAX3, Androgen receptor, Cyclin Bl, MYCN, RhoC, TRP-2, CYPIB I, BORIS, SART3, PAX5, OY- TES1, LCK, AKAP-4, SSX2, RAGE-1, human telomerase reverse transcriptase, RU1, RU2, intestinal carboxyl esterase, mut hsp70-2, CD79a, CD79b, CD72, LAIR1, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, GPC3, FCRL5, a neoantigen, CD 15, CD 184, CD56, CD26, CD29, CD44, HLA-A, HLA-B, HLA-C, (HLA- A,B,C) CD49f, CD151 CD340, CD200, tkrA, trkB, trkC, GPRC5D, IGLL1, IL-llRa, KLK2, KLK3, LY6G6D, p53, p53R175H, KRAS, KRAS G12D, PRAME, SIRPa, TAG72, ALPI, citrullinated vimentin, Axl, or an antigenic fragment or antigenic portion thereof.

[0420] In some embodiments, exemplary target antigens include, but are not limited to, CD1, CDla, CDlb, CDlc, CDld, CDle, CD2, CD3d, CD3e, CD3g, CD3s, CD4, CD5, CD7, CD8a, CD8b, CD19, CD20, CD21, CD22, CD23, CD24, CD25, CD27, CD28, CD30, CD33, CD34, CD35, CD38, CD40, CD44v6, CD45, CD45RA, CD45RO, CD46, CD47, CD48, CD52, CD56, CD59, CD62L, CD66, CD68, CD70, CD71, CD72, CD73, CD79A, CD79B, CD80 (B7.1), CD86 (B7.2), CD94, CD95, CD97, CD117, CD123, CD213A2, CD127, CD133, CD134, CD140 (PDGFR4), CD152, CD154, CD158, CD171, CD178, CD179, CD179a, CD181 (CXCR1), CD182 (CXCR2), CD183 (CXCR3), CD210, CD213A2, CD246, CD252, CD253, CD261, CD262, CD272, CD273 (PD-L2), CD274 (PD- Ll), CD276 (B7H3), CD279, CD295, CD339 (JAG1), CD340 (HER2), CDH17, CEA, CLL- 1, CS1, EGFR, FGFR2, AFP, CA125, MUC-1, MAGE, alkaline phosphatase, placental-like 2 (ALPPL2), B-cell maturation antigen (BCMA), green fluorescent protein (GFP), enhanced green fluorescent protein (eGFP), Claudinl8.2, PSMA, Mesothelin, IL13Ra2, FAP, signal regulatory protein a (SIRPa), TSHR, EGFRvIII, GD2, GD3, Tn Ag, ROR1, ROR2, GPC1, GPC2, FLT3, FAP, TAG72, CEA, EPCAM, B7H3, KIT, IL-13Ra2, IL-l lRa, PSCA, PRSS21, VEGFR2, LewisY, PDGFR-beta, SSEA-4, , folate receptor alpha, ERBB2 (Her2 / neu), MUC1, MUC16, EGFR, NCAM, prostase, PAP, ELF2M, Ephrin B2, IGF-1receptor, CAIX, LMP2, gplOO, bcr-abl, tyrosinase, EphA2, fucosyl GM1, sLe, GM3, TGS5, HMWMAA, o-acetyl-GD2, folate receptor beta, TEM1 / CD248, TEM7R, CLDN6, GPRC5D, CXORF61, ALK, Polysialic acid, PLAC1, GloboH, NY-BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP, WT1, NY-ESO-1, LAGE-la, MAGE- Al, legumain, HPV E6,E7, MAGE Al, ETV6-AML, sperm protein 17, XAGE1, Tie 2, MAD-CT-1, MAD-CT-2, Fos-related antigen 1, p53, p53 mutant, KRAS, mutant KRAS, KRAS G12D, prostein, surviving, telomerase, PC TA- 1 / Gal ectin 8, MelanA / MARTl, Ras mutant, hTERT, sarcoma translocation breakpoints, ML-IAP, ERG (TMPRSS2 ETS fusion gene), NA17, PAX3, androgen receptor, cyclin Bl, MYCN, RhoC, TRP-2, CYP1B1, BORIS, SART3, PAX5, OY-TES1, LCK, AKAP-4, SSX2, RAGE-1, human telomerase reverse transcriptase, RU1, RU2, intestinal carboxyl esterase, mut hsp70-2,LAIRl, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, GPC3, FCRL5, IGLL1, AFP, folate receptor alpha, ERBB2 (Her2 / neu), MUC1, MUC16, EGFR, NCAM, prostase, PAP, ELF2M, Ephrin B2, IGF -I receptor, CAIX, LMP2, gplOO, bcr-abl, tyrosinase, EphA2, fucosyl GM1, sLe, GM3, TGS5, HMWMAA, o-acetyl-GD2, folate receptor beta, TEM1 / CD248, TEM7R, CLDN6, GPRC5D, CXORF61, , ALK, Polysialic acid, PLAC1, GloboH, NY-BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP, WT1, NY-ESO-1, LAGE-la, MAGE- Al, legumain, HPV E6,E7, MAGE Al, ETV6-AML, sperm protein 17, XAGE1, Tie 2, MAD-CT-1, MAD-CT-2, Fos-related antigen 1, p53, p53 mutant, prostein, surviving, telomerase, PCTA-l / Galectin 8, MelanA / MARTl, Ras mutant, hTERT, sarcoma translocation breakpoints, ML-IAP, ERG (TMPRSS2 ETS fusion gene), NA17, PAX3, androgen receptor, cyclin Bl, MYCN, RhoC, TRP-2, CYP1B1, BORIS, SART3, PAX5, OY-TES1, LCK, AKAP-4, SSX2, RAGE-1, human telomerase reverse transcriptase, RU1, RU2, intestinal carboxyl esterase, mut hsp70-2, LAIR1, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, GPC3, FCRL5, IGLL1, the extracellular portion of the APRIL protein, or any combinations thereof.

[0421] In some embodiments, the target antigen is an antigen that includes, but is not limited to, DLL3, LY6G6D, CLDN6, GCC, p53R175H, KRAS G12D, PRAME, KLK2, KLK3, CD70, STEAP1, STEAP2, GPC3, TROP2, CLDN18.2, ROR1, ROPN1, IGF2BP3, CD19, BCMA, GPRC5D, CD20, CD22, CD79a, CD79b, GD2, GD3, and B7H3.

[0422] In some aspects, the target antigen is AFP, CD 19, TRAC, TCRP, BCMA, CLL-1, CS1, CD38, CD19, TSHR, CD123, CD22, CD30, CD171, CD33, EGFRvIII, GD2, GD3, Tn Ag, PSMA, ROR1, ROR2, GPC1, GPC2, FLT3, FAP, TAG72, CD44v6, CEA, EPCAM,B7H3, KIT, IL-13Ra2, IL-l lRa, PSCA, PRSS21, VEGFR2, LewisY, CD24, PDGFR-beta, S SEA-4, CD20, folate receptor alpha, ERBB2 (Her2 / neu), MUC1, MUC16, EGFR, NCAM, prostase, PAP, ELF2M, Ephrin B2, IGF-I receptor, CAIX, LMP2, gplOO, bcr-abl, tyrosinase, EphA2, fucosyl GM1, sLe, GM3, TGS5, HMWMAA, o-acetyl-GD2, folate receptor beta, TEM1 / CD248, TEM7R, CLDN6, GPRC5D, CXORF61, CD97, CD179a, ALK, Poly sialic acid, PLAC1, GloboH, NY-BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP, WT1, NY-ESO-1, LAGE-la, MAGE-A1, legumain, HPV E6,E7, MAGE Al, ETV6-AML, sperm protein 17, XAGE1, Tie 2, MAD-CT-1, MAD-CT-2, Fos-related antigen 1, p53, p53 mutant, KRAS, mutant KRAS, KRAS G12D, prostein, surviving, telomerase, PCTA-l / Galectin 8, MelanA / MARTl, Ras mutant, hTERT, sarcoma translocation breakpoints, ML-IAP, ERG (TMPRSS2 ETS fusion gene), NA17, PAX3, androgen receptor, cyclin Bl, MYCN, RhoC, TRP-2, CYP1B1, BORIS, SART3, PAX5, OY-TES1, LCK, AKAP-4, SSX2, RAGE-1, human telomerase reverse transcriptase, RU1, RU2, intestinal carboxyl esterase, mut hsp70-2, CD79a, CD79b, CD72, LAIR1, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, GPC3, FCRL5, IGLL1, the extracellular portion of the APRIL protein, or any combinations thereof.

[0423] In some aspects, the CAR or TCR targets AFP, CD 19, TRAC, TCRp, BCMA, CLL-1, CS1, CD38, CD19, TSHR, CD123, CD22, CD30, CD171, CD33, EGFRvIII, GD2, GD3, Tn Ag, PSMA, ROR1, ROR2, GPC1, GPC2, FLT3, FAP, TAG72, CD44v6, CEA, EPCAM, B7H3, KIT, IL-13Ra2, IL-l lRa, PSCA, PRSS21, VEGFR2, LewisY, CD24, PDGFR-beta, S SEA-4, CD20, folate receptor alpha, ERBB2 (Her2 / neu), MUC1, MUC16, EGFR, NCAM, prostase, PAP, ELF2M, Ephrin B2, IGF-I receptor, CAIX, LMP2, gplOO, bcr-abl, tyrosinase, EphA2, fucosyl GM1, sLe, GM3, TGS5, HMWMAA, o-acetyl-GD2, folate receptor beta, TEM1 / CD248, TEM7R, CLDN6, GPRC5D, CXORF61, CD97, CD 179a, ALK, Poly sialic acid, PLAC1, GloboH, NY-BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP, WT1, NY-ESO-1, LAGE-la, MAGE-A1, legumain, HPV E6,E7, MAGE Al, ETV6-AML, sperm protein 17, XAGE1, Tie 2, MAD- CT-1, MAD-CT-2, Fos-related antigen 1, p53, p53 mutant, KRAS, mutant KRAS, KRAS G12D, prostein, surviving, telomerase, PCTA-l / Galectin 8, MelanA / MARTl, Ras mutant, hTERT, sarcoma translocation breakpoints, ML-IAP, ERG (TMPRSS2 ETS fusion gene), NA17, PAX3, androgen receptor, cyclin Bl, MYCN, RhoC, TRP-2, CYP1B1, BORIS, SART3, PAX5, OY-TES1, LCK, AKAP-4, SSX2, RAGE-1, human telomerase reverse transcriptase, RU1, RU2, intestinal carboxyl esterase, mut hsp70-2, CD79a, CD79b, CD72,LAIR1, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, GPC3, FCRL5, IGLL1, the extracellular portion of the APRIL protein, or any combinations thereof.

[0424] In some embodiments, the CAR is a DLL3 CAR. In some embodiments, the extracellular binding domain of the DLL3 CAR comprises an antibody that binds to DLL3, for example, human DLL3. In some embodiments, the DLL3 CAR comprises any one of SEQ ID NOs: 401-413. In some embodiments, the extracellular binding domain of the DLL3 CAR comprises an antibody light chain sequence comprising SEQ ID NO: 415. In some embodiments, the extracellular binding domain of the DLL3 CAR comprises a antibody heavy chain sequence comprising SEQ ID NO: 417. In some embodiments, the extracellular binding domain of the DLL3 CAR comprises a light chain comprising a CDR1 comprising SEQ ID NO: 424, a CDR2 comprising SEQ ID NO: 425, and a CDR3 comprising SEQ ID NOs: 426. In some embodiments, the extracellular binding domain of the DLL3 CAR comprises a heavy chain comprising a CDR1 comprising SEQ ID NO: 427, a CDR2 comprising SEQ ID NO: 428, and a CDR3 comprising SEQ ID NOs: 429.

[0425] In some embodiments, the CAR is a LY6G6D CAR. In some embodiments, the extracellular binding domain of the LY6G6D CAR comprises an antibody that binds to LY6G6D, for example, human LY6G6D.

[0426] In some embodiments, the CAR is a CLDN6 CAR. In some embodiments, the extracellular binding domain of the CLDN6 CAR comprises an antibody that binds to CLDN6, for example, human CLDN6.

[0427] In some embodiments, the CAR is a CLDN18.2 CAR. In some embodiments, the extracellular binding domain of the CLDN18.2 CAR comprises an antibody that binds to CLDN18.2, for example, human CLDN18.2.

[0428] In some embodiments, the CAR is a GCC CAR. In some embodiments, the extracellular binding domain of the GCC CAR comprises an antibody that binds to GCC, for example, human GCC.

[0429] In some embodiments, the CAR is a PRAME CAR. In some embodiments, the extracellular binding domain of the PRAME CAR comprises an antibody that binds to PRAME, for example, human PRAME.

[0430] In some embodiments, the CAR is a KLK2 CAR. In some embodiments, the extracellular binding domain of the KLK2 CAR comprises an antibody that binds to KLK2, for example, human KLK2.

[0431] In some embodiments, the CAR is a KLK3 CAR. In some embodiments, the extracellular binding domain of the KLK3 CAR comprises an antibody that binds to KLK3, for example, human KLK3.

[0432] In some embodiments, the CAR is a CD70 CAR. In some embodiments, the extracellular binding domain of the CD70 CAR comprises an antibody that binds to CD70, for example, human CD70.

[0433] In some embodiments, the CAR is a STEAP1 CAR. In some embodiments, the extracellular binding domain of the STEAP1 CAR comprises an antibody that binds to STEAP1, for example, human STEAP1.

[0434] In some embodiments, the CAR is a STEAP2 CAR. In some embodiments, the extracellular binding domain of the STEAP2 CAR comprises an antibody that binds to STEAP2, for example, human STEAP2.

[0435] In some embodiments, the CAR is a GPC3 CAR. In some embodiments, the extracellular binding domain of the GPC3 CAR comprises an antibody that binds to GPC3, for example, human GPC3.

[0436] In some embodiments, the CAR is a TROP2 CAR. In some embodiments, the extracellular binding domain of the TROP2 CAR comprises an antibody that binds to TROP2, for example, human TROP2.

[0437] In some embodiments, the CAR is a ROPN1 CAR. In some embodiments, the extracellular binding domain of the ROPN1 CAR comprises an antibody that binds to ROPN1, for example, human ROPN1.

[0438] In some embodiments, the CAR is a ROR1 CAR. In some embodiments, the extracellular binding domain of the ROR1 CAR comprises an antibody that binds to ROR1, for example, human ROR1.

[0439] In some embodiments, the CAR is a IGF2BP3 CAR. In some embodiments, the extracellular binding domain of the IGF2BP3 CAR comprises an antibody that binds to IGF2BP3, for example, human IGF2BP3.

[0440] In some embodiments, the CAR is a CD79a CAR. In some embodiments, the extracellular binding domain of the CD79a CAR comprises an antibody that binds to CD79a, for example, human CD79a.

[0441] In some embodiments, the CAR is a CD79b CAR. In some embodiments, the extracellular binding domain of the CD79b CAR comprises an antibody that binds to CD79b, for example, human CD79b.

[0442] In some embodiments, the CAR is a GD2 CAR. In some embodiments, the extracellular binding domain of the GD2 CAR comprises an antibody that binds to GD2 for example, human GD2.

[0443] In some embodiments, the CAR is a GD3 CAR. In some embodiments, the extracellular binding domain of the GD3 CAR comprises an antibody that binds to GD3 for example, human GD3.

[0444] In some embodiments, the CAR is a B7H3 CAR. In some embodiments, the extracellular binding domain of the B7H3 CAR comprises an antibody that sbinds to B7H3 for example, human B7H3.

[0445] In some embodiments, the CAR is a CD 19 CAR. In some embodiments, the extracellular binding domain of the CD 19 CAR comprises an antibody that binds to CD 19, for example, human CD 19. In some embodiments, the extracellular binding domain of the CD 19 CAR comprises an scFv antibody fragment derived from the FMC63 monoclonal antibody (FMC63), which comprises the heavy chain variable region (VH) and the light chain variable region (VL) of FMC63 connected by a linker peptide. In some embodiments, the linker peptide is a "Whitlow" linker peptide. FMC63 and the derived scFv have been described in Nicholson et al., Mai. Immun. 34(16-17): 1157-1165 (1997) and PCT Application Publication No. WO2018 / 213337 A 1, the entire content of each of which is incorporated by reference herein. In some embodiments, the extracellular binding domain of the CD 19 CAR comprises an antibody derived from one of the CD19-specific antibodies including, for example, SJ25C1 (Bejcek et al., Cancer Res. 55:2346-2351 (1995)), HD37 (Pezutto et al., J. Immunol. 138(9):2793-2799 (1987)), 4G7 (Meeker et al., Hybridoma 3:305-320 (1984)), B43 (Bejcek (1995)), BLY3 (Bejcek (1995)), B4 (Freedman et al., 70:418-427 (1987)), B4 HB12b (Kansas & Tedder, J. Immunol. 147:4094-4102 (1991);Yazawa et al., Proc. Natl. Acad. Sci. USA 102: 15178- 15183 (2005); Herbst et al., J. Pharmacol. Exp. Ther. 335:213-222 (2010)), BU12 (Gallard et al., J. Immunology, 148(10): 2983-2987 (1992)), and CLB-CD19 (De Rie Cell. Immunol. 118:368- 381(1989)).

[0446] In some embodiments, the CAR is CD22 CAR. CD22, which is a transmembrane protein found mostly on the surface of mature B cells that functions as an inhibitory receptor for B cell receptor (BCR) signaling. CD22 is expressed in 60-70% of B cell lymphomas and leukemias (e.g., B-chronic lymphocytic leukemia, hairy cell leukemia, acute lymphocytic leukemia (ALL), and Burkitt's lymphoma) and is not present on the cell surface in early stages of B cell development or on stem cells. In some embodiments, the CD22 CARcomprises an extracellular binding domain that binds CD22, a transmembrane domain, an intracellular signaling domain, and / or an intracellular costimulatory domain. In some embodiments, the extracellular binding domain of the CD22 CAR comprises an scFv antibody fragment derived from the m971 monoclonal antibody (m971), which comprises the heavy chain variable region (VH) and the light chain variable region (VL) of m971 connected by a linker. In some embodiments, the extracellular binding domain of the CD22 CAR comprises an scFv antibody fragment derived from m971-L7, which an affinity matured variant of m971 with significantly improved CD22 binding affinity compared to the parental antibody m971 (improved from about 2 nM to less than 50 pM). In some embodiments, the scFv antibody fragment derived from m971-L7 comprises the VH and the VL of m971-L7 connected by a 3xG4S linker. In some embodiments, the extracellular binding domain of the CD22 CAR comprises immunotoxins HA22 or BL22. Immunotoxins BL22 and HA22 are therapeutic agents that comprise an scFv that binds CD22 fused to a bacterial toxin, and thus can bind to the surface of the cancer cells that express CD22 and kill the cancer cells. BL22 comprises a dsFv of an anti-CD22 antibody, RFB4, fused to a 38- kDa truncated form of Pseudomonas exotoxin A (Bang et al., Clin. Cancer Res., 11 : 1545- 50 (2005)). HA22 (CAT8015, moxetumomab pasudotox) is a mutated, higher affinity version of BL22 (Ho et al., J. Biol. Chem., 280(1): 607-17 (2005)). Suitable sequences of antigen binding regions of HA22 and BL22 that bind CD22 are disclosed in, for example, U.S. Patent Nos. 7,541,034; 7,355,012; and 7,982,011, which are hereby incorporated by reference in their entirety.

[0447] In some embodiments, the CAR is BCMA CAR. BCMA is a tumor necrosis family receptor (TNFR) member expressed on cells of the B cell lineage, with the highest expression on terminally differentiated B cells or mature B lymphocytes. BCMA is involved in mediating the survival of plasma cells for maintaining long-term humoral immunity. The expression of BCMA has been recently linked to a number of cancers, such as multiple myeloma, Hodgkin's and non-Hodgkin's lymphoma, various leukemias, and glioblastoma. In some embodiments, the BCMA CAR comprises an extracellular binding domain that binds BCMA, a transmembrane domain, an intracellular signaling domain, and / or an intracellular costimulatory domain. In some embodiments, the extracellular binding domain of the BCMA CAR comprises an antibody that binds to BCMA, for example, human BCMA. CARs directed to BCMA have been described in PCT Application Publication Nos. WO2016 / 014789, WO2016 / 014565, WO2013 / 154760, and WO 2015 / 128653. BCMA-binding antibodies are also disclosed in PCT Application Publication Nos. WO2015 / 166073 and W02014 / 068079. In some embodiments, the extracellular binding domain of the BCMA CAR comprises an scFv antibody fragment derived from a murine monoclonal antibody as described in Carpenter et al., Clin. Cancer Res. 19(8):2048-2060 (2013). In some embodiments, the scFv antibody fragment is a humanized version of the murine monoclonal antibody (Sommermeyer et al., Leukemia 31 :2191-2199 (2017)). In some embodiments, the extracellular binding domain of the BCMA CAR comprises single variable fragments of two heavy chains (VHH) that can bind to two epitopes of BCMA as described in Zhao et al., J. Hematol. Oneal. 11(1): 141 (2018). In some embodiments, the extracellular binding domain of the BCMA CAR comprises a fully human heavy-chain variable domain (FHVH) as described in Lam et al., Nat. Commun. 11 (1) :283 (2020).

[0448] In some embodiments, the CAR is a GPRC5D CAR. In some embodiments, the extracellular binding domain of the GPRC5D CAR comprises an antibody thatbinds to GPRC5D, for example, human GPRC5D.

[0449] In some embodiments, the CAR is a CD20 CAR. In some embodiments, the extracellular binding domain of the CD20 CAR comprises an antibody that binds to CD20, for example, human CD20.

[0450] In some embodiments, the antigen binding region targets an antigen characteristic of an autoimmune or inflammatory disorder. In some embodiments, the antigen binding region binds an antigen associated with an autoimmune or inflammatory disorder. In some instances, the antigen is expressed by a cell associated with an autoimmune or inflammatory disorder. In some embodiments, the autoimmune or inflammatory disorder is selected from chronic graft-vs-host disease (GVHD), lupus, arthritis, immune complex glomerulonephritis, goodpasture, uveitis, hepatitis, systemic sclerosis or scleroderma, type I diabetes, multiple sclerosis, cold agglutinin disease, Pemphigus vulgaris, Grave's disease, autoimmune hemolytic anemia, Hemophilia A, Primary Sjogren's Syndrome, thrombotic thrombocytopenia purrpura, neuromyelits optica, Evan's syndrome, IgM mediated neuropathy, cyroglobulinemia, dermatomyositis, idiopathic thrombocytopenia, ankylosing spondylitis, bullous pemphigoid, acquired angioedema, chronic urticarial, antiphospholipid demyelinating polyneuropathy, and autoimmune thrombocytopenia or neutropenia or pure red cell aplasias, while exemplary non-limiting examples of alloimmune diseases include allosensitization (see, for example, Blazar et al., 2015, Am. J. Transplant, 15(4):931-41) or xenosensitization from hematopoietic or solid organ transplantation, blood transfusions,pregnancy with fetal allosensitization, neonatal alloimmune thrombocytopenia, hemolytic disease of the newborn, sensitization to foreign antigens such as can occur with replacement of inherited or acquired deficiency disorders treated with enzyme or protein replacement therapy, blood products, and gene therapy. Allosensitization, in some instances, refers to the development of an immune response (such as circulating antibodies) against human leukocyte antigens that the immune system of the recipient subject or pregnant subject considers to be non-self antigens. In some embodiments, the antigen characteristic of an autoimmune or inflammatory disorder is selected from a cell surface receptor, an ion channel-linked receptor, an enzyme-linked receptor, a G protein-coupled receptor, receptor tyrosine kinase, tyrosine kinase associated receptor, receptor-like tyrosine phosphatase, receptor serine / threonine kinase, receptor guanylyl cyclase, or histidine kinase associated receptor.

[0451] In some embodiments, the antigen binding region targets an antigen characteristic of an infectious disease. In some embodiments, the antigen binding region binds an antigen associated with an infectious disease. In some instances, the antigen is expressed by a cell affected by an infectious disease. In some embodiments, wherein the infectious disease is selected from HIV, hepatitis B virus, hepatitis C virus, Human herpes virus, Human herpes virus 8 (HHV-8, Kaposi sarcoma- associated herpes virus (KSHV)), Human T- lymphotrophic virus-1 (HTLV-1), Merkel cell polyomavirus (MCV), Simian virus 40 (SV40), Epstein-Barr virus, CMV, human papillomavirus. In some embodiments, the antigen characteristic of an infectious disease is selected from a cell surface receptor, an ion channel-linked receptor, an enzyme-linked receptor, a G protein-coupled receptor, receptor tyrosine kinase, tyrosine kinase associated receptor, receptor-like tyrosine phosphatase, receptor serine / threonine kinase, receptor guanylyl cyclase, histidine kinase associated receptor, HIV Env, gpl20, or CD4-induced epitope on HIV-1 Env.

[0452] In any of these embodiments, the extracellular binding domain of the CAR can be codon-optimized for expression in a host cell or have variant sequences to increase functions of the extracellular binding domain.

[0453] In some embodiments, the CAR is bispecific to two target antigens (e.g., comprising binding region(s) for two target antigens). In some embodiments, the target antigens are different target antigens. In some of any such embodiments, the two different target antigens are any two different antigens described above. In some embodiments, the extracellular binding domains are different and bind two different antigens from (i) CD 19and CD20, (ii) CD20 and LI -CAM, (iii) LI -CAM and GD2, (iv) EGFR and LI -CAM, (v) CD 19 and CD22, (vi) EGFR and C-MET, (vii) EGFR and HER2, (viii) C-MET and HER2, or (ix) EGFR and ROR1. In some embodiments, each of the two different antigen binding regions is an scFv. In some embodiments, the C-terminus of one variable domain (VH or VL) of a first scFv is tethered to the N-terminus of the second scFv (VL or VH, respectively) via a polypeptide linker. In some embodiments, the linker connects the N-terminus of the VH with the C-terminus of VL or the C-terminus of VH with the N-terminus of VL. These scFvs lack the constant regions (Fc) present in the heavy and light chains of the native antibody. The scFvs, with capability for binding at least two different antigens, are arranged in tandem and linked to the co-stimulatory domain and the intracellular signaling domain via a transmembrane domain. In some embodiments, an extracelluar spacer domain may be linked between the antigen-binding region and the transmembrane domain.

[0454] In some embodiments, each antigen-binding region of the CAR comprises a divalent (or bivalent) single-chain variable fragment (di-scFvs, bi-scFvs). In CARs comprising di-scFVs, two scFvs with capability for binding each antigen are linked together by producing a single peptide chain with two VH and two VL regions, yielding tandem scFvs. (Xiong, Cheng- Yi; Natarajan, A; Shi, X B; Denardo, G L; Denardo, S J (2006). “Development of tumor targeting anti-MUC-1 multimer: effects of di-scFv unpaired cysteine location on PEGylation and tumor binding”. Protein Engineering Design and Selection 19 (8): 359-367; Kufer, Peter; Lutterbiise, Ralf; Baeuerle, Patrick A. (2004). “A revival of bispecific antibodies”. Trends in Biotechnology 22 (5): 238-244). CARs comprising at least two antigen-binding regions would express two scFvs with capability for binding each of the two antigens. The resulting antigen-binding region, with capability for binding at least two different antigens, is joined to the co-stimulatory domain and the intracellular signaling domain via a transmembrane domain. In some embodiments, an extracelluar spacer domain may be linked between the antigen binding domain and the transmembrane domain.

[0455] In some embodiments, each antigen-binding region of the CAR comprises a diabody. In a diabody, the scFvs are created with linker peptides that are too short for the two variable regions to fold together, driving the scFvs to dimerize. Still shorter linkers (one or two amino acids) lead to the formation of trimers, the so-called triabodies or tribodies. Tetrabodies may also be used.

[0456] In some embodiments, the cell is engineered to express more than one CAR, such as two different CARs, in which each CAR has an antigen binding region directed to a different target antigen. In some of any such embodiments, the two different target antigens are any two different antigens described above. In some embodiments, the extracellular binding domains are different and bind two different antigens from (i) CD 19 and CD20, (ii) CD20 and LI -CAM, (iii) LI -CAM and GD2, (iv) EGFR and LI -CAM, (v) CD 19 and CD22, (vi) EGFR and C-MET, (vii) EGFR and HER2, (viii) C-MET and HER2, or (ix) EGFR and ROR1.

[0457] In some embodiments, two different engineered cells are prepared that contain the provided modifications with each engineered with a different CAR. In some embodiments, each of the two different CARs has an antigen binding region directed to a different target antigen. In some of any such embodiments, the two different target antigens are any two different antigens described above. In some embodiments, the extracellular binding domains are different and bind two different antigens from (i) CD 19 and CD20, (ii) CD20 and LI - CAM, (iii) LI -CAM and GD2, (iv) EGFR and LI -CAM, (v) CD 19 and CD22, (vi) EGFR and C-MET, (vii) EGFR and HER2, (viii) C-MET and HER2, or (ix) EGFR and ROR1. In some embodiments, a population of engineered cells expressing a first CAR directed against a first target antigen and a population of engineered cells expressing a second CAR directed against a second target antigen are separately administered to the subject. In some embodiments, the first and second population of cells are administered sequentially in any order. For instance, the population of cells expressing the second CAR is administered a after administration of the population of cells expressing the first CAR.2. Linker Sequences

[0458] In some embodiments, the antigen binding region of the CAR may comprise one or more linker sequences. In some embodiments, the antigen binding region of the CAR may comprise one or more linker sequences connecting a heavy chain variable region (VH) and a light chain variable region (VL). The VH and the VL may be connected in either order, i.e., VH-linker-VL or VL-linker-VH. Nonlimiting examples of linkers include Whitlow linker, (G4S)n (n can be a positive integer, e.g., 1, 2, 3, 4, 5, 6, etc.) linker, and variants thereof.

[0459] Examples of exemplary linker sequences are described in Table 4. In provided aspects, the sequences of each linker in Table 4 can be combined with any of the other CAR component sequences described here.Table 4. Exemplary Linker Sequences3. Spacer (Hinge) Sequences

[0460] In some embodiments, the CAR further comprises one or more spacers, e.g., wherein the spacer is a first spacer between the antigen binding region and the transmembrane domain. In some embodiments, the first spacer includes at least a portion of an immunoglobulin constant region or variant or modified version thereof. In some embodiments, the spacer is a second spacer between the transmembrane domain and a signaling domain. In some embodiments, the second spacer is an oligopeptide, e.g., wherein the oligopeptide comprises glycine and serine residues such as but not limited to glycineserine doublets. In some embodiments, the CAR comprises two or more spacers, e.g., a spacer between the antigen binding region and the transmembrane domain and a spacer between the transmembrane domain and a signaling domain. The terms “hinge” and “spacer” may be used interchangeably in the present disclosure. Non-limiting examples of hinge domains include CD8a hinge domain, CD28 hinge domain, IgG4 hinge domain, IgG4 hinge-CH2-CH3 domain, and variants thereof.

[0461] Examples of exemplary spacer (hinge) sequences are described in Table 5. In provided aspects, the sequences of each spacer (hinge) in Table 5 can be combined with any of the other CAR component sequences described here.Table 5. Exemplary Spacer (Hinge) Sequences4. Transmembrane Domain

[0462] In some embodiments, the CAR transmembrane domain comprises at least a transmembrane region of the TCR alpha chain, TCR beta chain, TCR zeta chain, CD28, CD3 epsilon, CD3 zeta, l.xxCD3 zeta, CD45, CD4, CD5, CD8, CD9, CD 16, CD22, CD28, CD33, CD37, CD64, CD80, CD86, CD134, CD 137, CD 154, or functional variant thereof In some embodiments, the transmembrane domain comprises at least a transmembrane region(s) of CD8a, CD8P, 4-1BB / CD137, CD28, CD34, CD4, FcsRIy, CD16, OX40 / CD134, CD3< CD3s, CD3y, CD38, TCRa, TCRb, TCRA, CD32, CD64, CD64, CD45, CD5, CD9, CD22, CD37, CD80, CD86, CD40, CD40L / CD154, VEGFR2, FAS, and FGFR2B, or functional variant thereof.

[0463] Examples of exemplary transmembrane domain sequences are described in Table6. In provided aspects, the sequences of each transmembrane domain in Table 6 can be combined with any of the other CAR component sequences described here.Table 6. Exemplary Transmembrane Domain Sequences5. Signaling Domain(s)

[0464] In some embodiments, a CAR described herein comprises one or more signaling domains. A CAR may comprise a costimulatory signaling domain, e.g., to increase signaling potency. See U.S. Pat. Nos. 7,741,465, and 6,319,494, as well as Krause et al. and Finney et al. (supra), Song et al., Blood 119:696-706 (2012); Kalos et al, Sci Transl. Med. 3:95 (2011); Porter et al, N. Engl. J. Med. 365:725-33 (2011), and Gross et al, Annu. Rev. Pharmacol. Toxicol. 56:59-83 (2016). Signals generated through a TCR alone may be insufficient for full activation of a T cell and a secondary or co-stimulatory signal may increase activation. Thus, in some embodiments, a signaling domain further comprises one or more additional signaling domains (e.g., costimulatory signaling domains) that activate one or more immune cell effector functions (e.g., a native immune cell effector function described herein). In some embodiments, a portion of such costimulatory signaling domains may be used, as long as the portion transduces the effector function signal. In some embodiments, a cytoplasmic domain described herein comprises one or more cytoplasmic sequences of a T cell coreceptor (or fragment thereof). Non-limiting examples of such T cell co-receptors comprise CD27, CD28, 4-1BB (CD137), 0X40, CD30, CD40, PD-1, ICOS, lymphocyte function- associated antigen-1 (LFA- 1), MYD88, CD2, CD7, LIGHT, NKG2C, B7-H3, and a ligand that binds with CD83. An exemplary costimulatory protein has the amino acid sequence of acostimulatory protein found naturally on T cells, the complete native amino acid sequence of which costimulatory protein is described in NCBI Reference Sequence: NP 006130.1.

[0465] In some embodiments, a CAR described herein comprises one or more signaling domain selected from one or more of B7-1 / CD80; B7-2 / CD86; B7-H1 / PD-L1; B7-H2; B7- H3; B7-H4; B7-H6; B7-H7; BTLA / CD272; CD28; CTLA-4; Gi24 / VISTA / B7-H5;ICOS / CD278; PD-1; PD-L2 / B7-DC; PDCD6); 4-1BB / TNFSF9 / CD137; 4-1BB Ligand / TNFSF9; BAFF / BLyS / TNFSF13B; BAFF R / TNFRSF13C; CD27 / TNFRSF7; CD27 Ligand / TNFSF7; CD30 / TNFRSF8; CD30 Ligand / TNFSF8; CD40 / TNFRSF5;CD40 / TNFSF5; CD40 Ligand / TNFSF5; DR3 / TNFRSF25; GITR / TNFRSF18; GITR Ligand / TNFSF18; HVEM / TNFRSF14; LIGHT / TNFSF14; Lymphotoxin-alpha / TNF-beta; OX40 / TNFRSF4; 0X40 Ligand / TNFSF4; RELT / TNFRSF19L; TACI / TNFRSF13B;TL1A / TNFSF15; TNF-alpha; TNF RII / TNFRSF1B); 2B4 / CD244 / SLAMF4; BLAME / SLAMF8; CD2; CD2F-10 / SLAMF9; CD48 / SLAMF2; CD58 / LFA-3; CD84 / SLAMF5; CD229 / SLAMF3; CRACC / SLAMF7; NTB- A / SLAMF6; SLAM / CD150); CD2; CD7; CD53; CD82 / Kai-1; CD90 / Thyl; CD96; CD160; CD200; CD300a / LMIRl; HLA Class I; HLA-DR; Ikaros; Integrin alpha 4 / CD49d; Integrin alpha 4 beta 1; Integrin alpha 4 beta 7 / LPAM-l; LAG-3; TCL1A; TCL1B; CRTAM; DAP12; Dectin- 1 / CLEC7A;DPPIV / CD26; EphB6; TIM- 1 / KIM- 1 / HA VCR; TIM-4; TSLP; TSLP R; lymphocyte function associated antigen-1 (LFA-1); NKG2C, a CD3 zeta domain, a l.xxCD3 zeta domain, an immunoreceptor tyrosinebased activation motif (ITAM), CD27, CD28, 4-1BB, CD134 / OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA- 1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that binds CD83, or functional fragment thereof.

[0466] In some embodiments, a CAR comprises a signaling domain which is a costimulatory domain. In some embodiments, a CAR comprises a second costimulatory domain. In some embodiments, a CAR comprises at least two costimulatory domains. In some embodiments, a CAR comprises at least three costimulatory domains. In some embodiments, a CAR comprises a costimulatory domain selected from one or more of CD3zeta, LxxCD3zeta, CD27, CD28, 4-1BB, CD134 / OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that binds CD83. In some embodiments, if a CAR comprises two or more costimulatory domains, two costimulatory domains are different. In some embodiments, if aCAR comprises two or more costimulatory domains, two costimulatory domains are the same.

[0467] In other embodiments, the at least one signaling domain comprises (i) a 4- IBB domain, or functional variant thereof; and (ii) a CD3 zeta domain, or functional variant thereof. In other embodiments, the at least one signaling domain comprises (i) a CD28 domain, or functional variant thereof; and (ii) a CD3 zeta domain, or functional variant thereof. In other embodiments, the at least one signaling domain comprises (i) a CD28 domain, or functional variant thereof; and (ii) a 1 ,xxCD3 zeta domain, or functional variant thereof.

[0468] In some embodiments, the CAR comprises an extracellular antigen binding region (e.g., antibody or antibody fragment, such as an scFv) that binds to an antigen (e.g., tumor antigen), a spacer (e.g., containing a hinge domain, such as any as described herein), a transmembrane domain (e.g., any as described herein), and one or more intracellular signaling domains (e.g., any intracellular signaling domain, such as a primary signaling domain or costimulatory signaling domain as described herein). In some embodiments, the one or more intracellular signaling domains is or comprises a primary cytoplasmic signaling domain. In some embodiments, the one or more intracellular signaling domains additionally comprises an intracellular signaling domain of a costimulatory molecule (e.g., a costimulatory domain). Any of such components can be any as described above.

[0469] Examples of exemplary signaling domain sequences are described in Table 7. In provided aspects, the sequences of each signaling domain in Table 7 can be combined with any of the other CAR component sequences described here.Table 7. Exemplary Signaling Domain Sequences6. Signal Peptide

[0470] In certain embodiments, the CAR may further comprise one or more signal peptides. In certain embodiments, the CAR may comprise a signal peptide at the N-terminus. Non-limiting examples of signal peptides include CD8a signal peptide, IgK signal peptide, and granulocyte-macrophage colony-stimulating factor receptor subunit alpha (GMCSFR-a, also known as colony stimulating factor 2 receptor subunit alpha (CSF2RA)) signal peptide, and variants thereof, the amino acid sequences of which are provided in Table 8 below.

[0471] Examples of exemplary signal peptide sequences are described in Table 8. In provided aspects, the sequences of each signal peptide in Table 8 can be combined with any of the other CAR component sequences described here.Table 8. Exemplary Signal Peptide Sequencesb. T Cell Receptors (TCRs)

[0472] In some embodiments, the receptor of the disclosure is a T cell receptor (TCR). A TCR as known by the skilled in the art can be composed of two different, an alpha chain and a beta chain, each consisting of a constant region that anchors the chain inside the T cell surface membrane, and a variable region which can recognize and bind an antigen presented by MHCs. The TCR complex can be associated with six polypeptides forming two heterodimers, CD3ys and CD35s, and one homodimer, CD3(^, which together forms the CD3 complex. TCRs can be engineered to utilize the modification of T cells that retain these complexes to bind the antigens expressed by particular tumor cells. As used herein, a TCR can be a naturally-occurring or an engineered TCR.

[0473] TCRs are disulfide-linked membrane anchored heterodimeric proteins, typically comprising highly variable alpha (a) and beta (P) chains expressed as a complex with invariant CD3 chain molecules. T cells expressing these types of TCRs are referred to as a:P (or aP) T cells. A minority of T cells express an alternative TCR comprising variable gamma (y) and delta (5) chains and are referred to as y5 T cells. TCR is not able to mediate signal transduction itself due to its short cytoplasmic tail, so TCR still requires CD3 and zeta to carry out the signal transduction in its place. A TCR receptor complex is an octomeric complex of variable TCR receptor a and P chains with three dimeric signaling modules

[0474] According to embodiments of the application, suitable TCRs are capable of binding a major histocompatibility complex (MHC) on the surface of cancer cells that displays a peptide fragment of a tumor antigen. An MHC is a set of cell-surface proteins which allow the acquired immune system to recognize ‘foreign’ molecules. Proteins are intracellularly degraded and presented on the surface of cells by the MHC. MHCs displaying “foreign” peptides, such a viral or cancer associated peptides, are recognized by T cells with the appropriate TCRs, prompting cell destruction pathways. MHCs on the surface of cancer cells can display peptide fragments of tumor antigen i.e. an antigen which is present on a cancer cell but not the corresponding non-cancerous cell. T cells which recognize these peptide fragments can exert a cytotoxic effect on the cancer cell.

[0475] According to embodiments of the application, the T cells can be modified to express a heterologous TCR that is capable of binding to MHCs displaying peptide fragments of a tumor antigen expressed by the cancer cells in a specific cancer patient. Tumor antigens expressed by cancer cells in the cancer patient may be identified using standard techniques. For example, a polypeptide, such as an FSH fragment, which is capable of binding to an FSHR, is fused to CD3 epsilon, CD3 gamma or CD3 delta chain. The fusion protein forms a TCR complex on T cells through the interaction of CD3 with TCR a / p chains. The TCR complex is capable of binding FSHR on tumor cells via the FSH fragment, and the binding initiates TCR signaling against the tumor cells. According to yet another embodiment of the invention, the heterologous TCR is expressed together with a CAR that is capable of binding mesothelin. Heterologous TCRs can include aPTCR heterodimers.

[0476] The TCR can be engineered to increase its affinity or avidity for a tumor antigen (i.e., an affinity enhanced TCR). The affinity enhanced TCR can comprise one or moremutations relative to a naturally occurring TCR, for example, one or more mutations in the hypervariable complementarity determining regions (CDRs) of the variable regions of the TCR a and P chains. These mutations increase the affinity of the TCR for MHCs that display a peptide fragment of a tumor antigen expressed by cancer cells. Suitable methods of generated affinity enhanced TCRs include screening libraries of TCR mutants using phage or yeast display and are well known in the art (see for example Robbins et al J Immunol (2008) 180 (9) : 6116; San Miguel et al (2015) Cancer Cell 28 (3) 281-283; Schmitt et al (2013) Blood 122 348-256; Jiang et al (2015) Cancer Discovery 5 901).

[0477] In some of embodiments, the cell comprises a recombinant nucleic acid encoding a TCRa and / or TCRb chain or a portion thereof that has been codon-optimized. In some embodiments, the transgene encodes a portion of a TCRa and / or TCRb chain with less than 100% amino acid sequence identity to a corresponding portion of a native or endogenous TCRa and / or TCRb chain. In some embodiments, the encoded TCRa and / or TCRb chain contains an amino acid sequence with, with about, or with at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or greater than 99% identity but less than 100% identity to a corresponding native or endogenous TCRa and / or TCRb chain. In particular embodiments, the transgene encodes a TCRa and / or TCRb constant domain or portion thereof with less than 100% amino acid sequence identity to a corresponding native or endogenous TCRa and / or TCRb constant domain. In some embodiments, the TCRa and / or TCRb constant domain contains an amino acid sequence with, with about, or with at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or greater than 99% identity but less than 100% identity to a corresponding native or endogenous TCRa and / or TCRb chain.

[0478] In some embodiments, the recombinant nucleic acid contains one or more modifications(s) to introduce one or more cysteine residues that are capable of forming one or more non-native disulfide bridges between the TCRa chain and TCRB chain. In some embodiments, the one or more non-native cysteine residues are capable of forming nonnative disulfide bonds, e.g., with a TCRa chain encoded by the transgene.

[0479] In some embodiments, the transgene encodes all or a portion of a TCRa constant domain (Ca) and / or a TCRb constant domain (Cb) with one or more modifications to remove or prevent a native disulfide bond, e.g., between the TCRb chain encoded by the transgene and the endogenous TCRa chain. In some embodiments, one or more native cysteines that form and / or are capable of forming a native interchain disulfide bond are substituted to another residue, e.g., serine or alanine. In some embodiments, the portion of aTCRa constant domain (Ca) and / or a TCRb constant domain (Cb) is modified to replace one or more non-cysteine residues to a cysteine. Exemplary of modified encoded TCRa constant domain (Ca) and / or TCRb constant domain (Cb) include any of those described herein, for example, in Section III. A, and those described in W02006 / 000830, WO 2006 / 037960 and Kuball et al. (2007) Blood, 109:2331- 2338.

[0480] Among such recombinant TCRs or antigen-binding fragments thereof are TCRs or antigen-binding fragments thereof that contain any of the variable alpha (Va) regions and / or a variable beta (Vb) regions as described, individually, or a sufficient antigen-binding portion of such chain(s). In some embodiment, the provided recombinant TCRs comprise a TCR alpha (TCRa) chain comprising a variable alpha (Va) region, and a TCR beta (TCRb) chain comprising a variable beta (Vb) region.

[0481] In some embodiments, the TCR or antigen-binding fragment thereof is isolated or purified or is recombinant. In particular embodiments, any of the provided TCR or antigenbinding fragment thereof is recombinant. In some aspects, the TCR or antigenbinding fragment thereof is human. In some embodiments, the recombinant TCR is monoclonal. In some aspects, the recombinant TCR is a single chain. In other embodiments, the recombinant TCR contains two chains. In some embodiments, the TCR or antigenbinding fragment thereof is expressed on the surface of a cell.

[0482] Unless otherwise stated, the term “TCR” should be understood to encompass full TCRs as well as antigen-binding portions or antigen-binding fragments thereof. In some embodiments, the TCR is an intact or full-length TCR, such as a TCR containing the a chain and b chain. In some embodiments, the TCR is an antigen-binding portion that is less than a full- length TCR but that binds to a specific peptide bound in an MHC molecule, such as binds to an MHC -peptide complex. In some cases, an antigen-binding portion or fragment of a TCR can contain only a portion of the structural domains of a full-length or intact TCR, but yet is able to bind the peptide epitope, such as MHC-peptide complex, to which the full TCR binds. In some cases, an antigen-binding portion contains the variable domains of a TCR, such as variable a (Va) chain and variable b (Vb) chain of a TCR, or antigen-binding fragments thereof sufficient to form a binding site for binding to a specific MHC-peptide complex.

[0483] In some embodiments, the variable domains of the TCR contain complementarity determining regions (CDRs), which generally are the primary contributors to antigen recognition and binding capabilities and specificity of the peptide, MHC and / or MHC-peptide complex. In some embodiments, a CDR of a TCR or combination thereof forms all or substantially all of the antigen-binding site of a given TCR molecule. The various CDRs within a variable region of a TCR chain generally are separated by framework regions (FRs), which generally display less variability among TCR molecules as compared to the CDRs (see, e.g., lores et al., Proc. Nat'l Acad. Sci. U.S.A. 57:9138, 1990; Chothia et al., EMBO J. 737A5, 1988; see also Lefranc et al., Dev. Comp. Immunol. 27:55, 2003). In some embodiments, CDR3 is the main CDR responsible for antigen binding or specificity, or is the most important among the three CDRs on a given TCR variable region for antigen recognition, and / or for interaction with the processed peptide portion of the peptide-MHC complex. In some contexts, the CDR1 of the alpha chain can interact with the N-terminal part of certain antigenic peptides. In some contexts, CDR1 of the beta chain can interact with the C-terminal part of the peptide. In some contexts, CDR2 contributes most strongly to or is the primary CDR responsible for the interaction with or recognition of the MHC portion of the MHC -peptide complex. In some embodiments, the variable region of the b-chain can contain a further hypervariable region (CDR4 or HVR4), which generally is involved in superantigen binding and not antigen recognition (Kotb (1995) Clinical Microbiology Reviews, 8:411-426).

[0484] In some embodiments, the TCRa chain and / or TCRb chain also can contain a constant domain, a transmembrane domain and / or a short cytoplasmic tail (see, e.g., Janeway et al., Immunobiology: The Immune System in Health and Disease, 3rdEd., Current Biology Publications, p. 4:33, 1997). In some aspects, each chain (e.g. alpha or beta) of the TCR can possess one N-terminal immunoglobulin variable domain, one immunoglobulin constant domain, a transmembrane region, and a short cytoplasmic tail at the C-terminal end. In some embodiments, a TCR, for example via the cytoplasmic tail, is associated with invariant proteins of the CD3 complex involved in mediating signal transduction. In some cases, the structure allows the TCR to associate with other molecules like CD3 and subunits thereof. For example, a TCR containing constant domains with a transmembrane region may anchor the protein in the cell membrane and associate with invariant subunits of the CD3 signaling apparatus or complex. The intracellular tails of CD3 signaling subunits (e.g. CD3y, CD36, CD3s and CD3(Achains) contain one or more immunoreceptor tyrosine-based activation motif or IT AM and generally are involved in the signaling capacity of the TCR complex.

[0485] The various domains or regions of a TCR can be identified. In some cases, the exact locus of a domain or region can vary depending on the particular structural or homology modeling or other features used to describe a particular domain. It is understood that reference to amino acids, including to a specific sequence set forth as a SEQ ID NO used to describe domain organization of a TCR are for illustrative purposes and are not meant to limit the scope of the embodiments provided. In some cases, the specific domain (e.g. variable or constant) can be several amino acids (such as one, two, three or four) longer or shorter. In some aspects, residues of a TCR are known or can be identified according to the International Immunogenetics Information System (IMGT) numbering system (see e.g. www.imgt.org; see also, Lefranc et al. (2003) Developmental and Comparative Immunology, 27(l);55-77; and The T Cell Factsbook 2nd Edition, Lefranc and LeFranc Academic Press 2001).

[0486] In some embodiments, the TCRa chain and / or TCRb chain each further contain a constant domain. In some embodiments, the a chain constant domain (Ca) and b chain constant domain (Cb) individually are mammalian, such as is a human or murine constant domain. In some embodiments, the constant domain is adjacent to the cell membrane. For example, in some cases, the extracellular portion of the TCR formed by the two chains contains two membrane-proximal constant domains, and two membrane-distal variable domains, which variable domains each contain CDRs.

[0487] In some aspects, provided herein are TCRs that contains a human constant region, such as an alpha chain containing a human Ca region and a beta chain containing a human Cb. In some embodiments, the provided TCRs are fully human. Among the provided TCRs are TCRs containing a human constant region, such as fully human TCRs, whose expression and / or activity, such as when expressed in human cells, e.g. human T cells, such as primary human T cells, are not impacted by or are not substantially impacted by the presence of an endogenous human TCR.

[0488] In some embodiments, the TCR may be a heterodimer of two chains TCRa and TCRb that are linked, such as by a disulfide bond or disulfide bonds. In some embodiments, the constant domain of the TCR may contain short connecting sequences in which a cysteine residue forms a disulfide bond, thereby linking the two chains of the TCR. In some embodiments, a TCR may have an additional cysteine residue in each of the TCRa and TCRb chains, such that the TCR contains two disulfide bonds in the constant domains. Insome embodiments, each of the constant and variable domains contains disulfide bonds formed by cysteine residues.

[0489] In some embodiments, the TCR can contain an introduced disulfide bond or bonds. In some embodiments, the native disulfide bonds are not present. In some embodiments, the one or more of the native cysteines (e.g. in the constant domain of the a chain and b chain) that form a native interchain disulfide bond are substituted to another residue, such as to a serine or alanine. In some embodiments, an introduced disulfide bond can be formed by mutating noncysteine residues on the alpha and beta chains, such as in the constant domain of the a chain and b chain, to cysteine. Opposing cysteines in the TCRa and TCRb chains provide a disulfide bond that links the constant regions of TCRa and TCRb chains of the substituted TCR to one another and which is not present in a TCR comprising the unsubstituted constant region in which the native disulfide bonds are present, such as unsubstituted native human constant region or the unsubstituted native mouse constant region. In some embodiments, the presence of non-native cysteine residues (e.g. resulting in one or more non-native disulfide bonds) in a recombinant TCR can favor production of the desired recombinant TCR in a cell in which it is introduced over expression of a mismatched TCR pair containing a native TCR chain. In some embodiments, the TCRa and / or TCRb chain and / or a TCRa and / or TCRb chain constant domains are modified to replace one or more non-cysteine residues to a cysteine. c. T cell receptor fusion constructs (TRuCs)

[0490] In some embodiments, the receptor of the disclosure is a T cell receptor fusion construct (TRuC) such as those described in WO2016187349A1, which is herein incorporated by reference in its entirety. The TRuC may comprise a TCR subunit and an antigen binding region. In some embodiments, the TRuC is able to associate with one or more endogenous (or alternatively, one or more exogenous, or a combination of endogenous and exogenous) TCR subunits in order to form a functional TCR complex. In some embodiments, the TRuC-mediated T-cell response is directed to the antigen that may be bound by the antigen binding region.[049...

Claims

CLAIMS1. A recombinant polypeptide comprising (i) an extracellular region comprising a binding region for an antigen; and (ii) a transmembrane region, wherein the extracellular region comprises a peptide epitope capable of being bound by an anti-CCR4 antibody, wherein the recombinant polypeptide is not CCR4.

2. The recombinant polypeptide of claim 1, wherein the recombinant polypeptide is a chimeric antigen receptor (CAR) or a recombinant T cell receptor (TCR).

3. The recombinant polypeptide of claim 2, wherein the anti-CCR4 antibody is mogamulizumab, wherein the peptide epitope consists of SEQ ID NO: 431, and wherein the recombinant polypeptide does not comprise a cysteine immediately after the C-terminus of the peptide epitope.

4. The recombinant polypeptide of claim 3, wherein the binding region comprises a heavy chain variable region (VH) and a light chain variable region (VL), and wherein the peptide epitope or mimotope is located between the VH and the VL.

5. An engineered immune cell expressing the recombinant polypeptide of any one of claims 1-4.

6. A method of treating cancer or an autoimmune disease in a subject in need thereof, comprising administering an effective amount of the engineered cells of claim 5 to the subject.

7. The method of claim 6, wherein the method further comprises administering the anti- CCR4 antibody to the subject, wherein the anti-CCR4 antibody is mogamulizumab.

8. The method of claim 7, wherein the anti-CCR4 antibody is administered after detection or occurrence of an adverse event caused by the engineered cells in the subject,and wherein administration of the anti-CCR4 antibody reduces or eliminates at least one symptom of the adverse event.

9. The method of claim 8, wherein administration of the anti-CCR4 antibody depletes at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 99%, at least 99.5%, or at least 99.9% of the engineered cells in the subject.

10. A modified peptide epitope or mimotope, wherein the peptide epitope comprises or consists of an amino acid sequence having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to SEQ ID NO: 430, and wherein the peptide epitope is capable of being bound by mogamulizumab.

11. The modified peptide epitope or mimotope of claim 10, wherein the modified peptide epitope or mimotope does not comprise the cysteine that is present at the C-terminus of SEQ ID NO: 430.

12. The modified peptide epitope or mimotope of claim 10 or 11, wherein the modified peptide epitope comprises or consists of any one of SEQ ID NO: 431-438; preferably, the modified peptide epitope consists of SEQ ID NO: 431.

13. The modified peptide epitope or mimotope of claim 10, wherein the peptide epitope or mimotope comprises or consists of an amino acid sequence:(a) having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to SEQ ID NO: 475, and wherein the peptide epitope or mimotope is capable of being bound by rituximab;(b) having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to SEQ ID NO: 476, and wherein the peptide epitope or mimotope is capable of being bound by Palivizumab;(c) having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to SEQ ID NO: 477, and wherein the peptide epitope or mimotope is capable of being bound by Cetuximab;(d) having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to SEQ ID NO: 478, and wherein the peptide epitope or mimotope is capable of being bound by Cetuximab;(e) having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to SEQ ID NO: 479, and wherein the peptide epitope or mimotope is capable of being bound by Cetuximab;(f) having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to SEQ ID NO: 480, and wherein the peptide epitope or mimotope is capable of being bound by Cetuximab;(g) having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to SEQ ID NO: 481, and wherein the peptide epitope or mimotope is capable of being bound by Nivolumab;(h) having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to SEQ ID NO: 482, and wherein the peptide epitope or mimotope is capable of being bound by Nivolumab;(i) having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to SEQ ID NO: 483, and wherein the peptide epitope or mimotope is capable of being bound by QBend-10;(j) having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to SEQ ID NO: 484, and wherein the peptide epitope or mimotope is capable of being bound by Alemtuzumab;(k) having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to SEQ ID NO: 469, and wherein the peptide epitope or mimotope is capable of being bound by Sacituzumab;(l) having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to any one of SEQ ID NO:470-474, and wherein the peptide epitope or mimotope is capable of being bound by an anti-GUCY2C antibody;(m) having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to SEQ ID NO: 439, and wherein the peptide epitope or mimotope is capable of being bound by Tarlatamab;(n) having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to any one of SEQ ID NO: 441-468, and wherein the peptide epitope or mimotope is capable of being bound by an anti-DLL3 antibody; or(o) having 1, 2, 3, 4 or 5 mutations (substitutions, insertions, and / or deletions) compared to any one of SEQ ID NOs: 519-524, and wherein the peptide epitope or mimotope is capable of being bound by CAP- 100.

14. The modified peptide epitope or mimotope of any one of claims 10-13, wherein the mutation(s) are substitutions.

15. The modified peptide epitope or mimotope of any one of claims 10-14, wherein the mutation(s) are conservative substitutions.

16. The modified peptide epitope or mimotope of any one of claims 10-15, wherein the cysteine(s) in the unmodified peptide epitope or mimotope are substituted or deleted.

17. The modified peptide epitope or mimotope of claim 16, wherein the N-terminus of the unmodified peptide epitope or mimotope contains cysteine.

18. The modified peptide epitope or mimotope of claim 16 or 17, wherein the C- terminus of the unmodified peptide epitope or mimotope contains cysteine.

19. The modified peptide epitope or mimotope of any one of claims 16-18, wherein the unmodified peptide epitope or mimotope comprises at least one cysteine between the N- terminus and the C-terminus.

20. The modified peptide epitope or mimotope of any one of claims 16-19, wherein the cysteine(s) in the unmodified peptide epitope or mimotope are substituted with serine, glycine, threonine, alanine, or valine; preferably, serine or valine; more preferably, serine.

21. The modified peptide epitope or mimotope of claim 20, wherein the cysteine(s) in the unmodified peptide epitope or mimotope are substituted with serine.

22. The modified peptide epitope or mimotope of claim 20, wherein the cysteine(s) in the unmodified peptide epitope or mimotope are substituted with valine.

23. The modified peptide epitope or mimotope of any one of claims 10-22, wherein the modified peptide epitope or mimotope does not comprise any sulfur atoms.

24. The modified peptide epitope or mimotope of any one of claims 10-23, wherein the modified peptide epitope or mimotope does not comprise any cysteine(s).

25. The modified peptide epitope or mimotope of any one of claims 10-24, wherein the mutation(s) comprise deleting or substituting one or more amino acids that facilitate polypeptide crosslinking in the unmodified peptide epitope or mimotope.

26. The modified peptide epitope or mimotope of any one of claims 10-25, wherein the mutation(s) comprise deleting or substituting one or more cysteine(s), lysine(s), aspartic acid(s), glutamic acid(s), or any combination thereof.

27. The modified peptide epitope or mimotope of any one of claims 10-26, wherein the modified peptide epitope or mimotope is capable of being bound by the corresponding antibody with a dissociation constant (Kd) of less than 1 pM, less than 100 nM, less than 10 nM, or less than 1 nM; optionally wherein the Kd is measured by surface plasmon resonance (SPR) using a sensor chip that contains the immobilized antibody.

28. The modified peptide epitope or mimotope of any one of claims 10-27, wherein the modified peptide epitope or mimotope is capable of being bound by [fam-]trastuzumab deruxtecan, (fam-trastuzumab deruxtecan-nxki), Adebrelimab, Ado-trastuzumab emtansine, Alemtuzumab, Amivantamab, Atezolizumab, Avelumab, Belantamab mafodotin (belantamab mafodotin-blmf), Bevacizumab, Blinatumomab, Brentuximab vedotin, Cadonilimab, Camrelizumab, CAP- 100, Catumaxomab, Cemiplimab (cemiplimab-rwlc), Cetuximab, Cetuximab saratolacan, Cosibelimab, Daratumumab, Dinutuximab, Disitamab vedotin, Dostarlimab, Durvalumab, Edrecolomab, Elotuzumab, Elranatamab, Emapalumab (emapalumab-lzsg), Enfortumab vedotin (enfortumab vedotin-ejfv), Enlonstobart, Envafolimab, Epcoritamab, Gemtuzumab ozogamicin, Glofitamab, Ibritumomab tiuxetan, Inotuzumab ozogamicin, Ipilimumab, Isatuximab (isatuximab-irfc), Loncastuximab tesirine, Margetuximab-cmkb, Mirvetuximab soravtansine, Mogamulizumab (mogamulizumab- kpkc), Mosunetuzumab, Moxetumomab pasudotox (moxetumomab pasudotox-tdfk),Naxitamab-gqgk, Necitumumab, Nimotuzumab, Nivolumab, Cipterbin, Obinutuzumab, Ofatumumab, Olaratumab, Panitumumab, Pembrolizumab, Penpulimab, Pertuzumab, Polatuzumab vedotin (polatuzumab vedotin-piiq), Prolgolimab, Pucotenlimab, Racotumomab, Ramucirumab, Relatlimab, Retifanlimab, Ripertamab, Rituximab, Sacituzumab govitecan (sacituzumab govitecan-hziy), Serplulimab, Sintilimab, Socazolimab, Sugemalimab, Tafasitamab (tafasitamab-cxix), Tagitanlimab, Talquetamab (talquetamab-tgvs), Tebentafusp, Teclistamab, Tislelizumab, Tisotumab vedotin, tisotumab vedotin-tftv, Toripalimab, Tositumomab-1131, Trastuzumab, Trastuzumab duocarmazine, Tremelimumab, Zimberelimab, Zolbetuximab, Zuberitamab, Odronextamab, Ivonescimab, Benmel Stobart, Trastuzumab botidotin, Iparomlimab, tuvonralimab, Tarlatamab, Patritumab deruxtecan, Sacituzumab tirumotecan, Zanidatamab, Linvoseltamab, Datopotamab deruxtecan, Anbenitamab, Anvatabart opadotin, Apamistamab-Iodine (1311), Bemarituzumab, Cetrelimab, Cobolimab, Domvanalimab, Emactuzumab, Erfonrilimab, Favezelimab, Felzartamab, Fianlimab, Finotonlimab, ABBV-383, Geptanolimab, Gotistobart, Ivuxolimab, Izalontamab, Lemzoparlimab, Luveltamab tazevibulin, Magrolimab, Mecbotamab vedotin, Monalizumab, Nofazinlimab, Nurulimab, Ociperlimab, Oleclumab, Onfekafusp alfa, Oregovomab, Ozekibart, Pivekimab sunirine, Quavonlimab, Retlirafusp alfa, Rosopatamab (177Lu-DOTA), Rulonilimab, Sasanlimab, Telisotuzumab vedotin, Tiragolumab, IKS014, LCB14-0110, FS-1502, Trastuzumab mafodotin, Trastuzumab rezetecan, Vibostolimab, Vobramitamab duocarmazine, Zilovertamab vedotin, Suvemcitug, APX003, BD0801, TK001, sevacizumab, CTX-009, ESI 04, TR009, NOV1501, ABL001, DP303c, IAH0968, Kintuximab, gentuximab, cintuximab, MIL62, MRG002, MRG003, PM8002, MEDI5752, MCLA-128, AGEN1181, L19-IL2, L19-TNF, IMC-F106C, JMT101, BL-B01D1, BAT1308, AZD2936, JS004, TAB004, FG-M108, M108, 9MW2821, BMS-986349, CC-93269, EM801, ABBV-181, ABBV-151, ARGX-115, DS-7300a, IBI343, SHR-A1921, or ASKB589.

29. The modified peptide epitope or mimotope of any one of claims 10-28, wherein the peptide epitope or mimotope comprises or consists of an amino acid sequence having 1, 2, 3, 4, or 5 mutations compared to the unmodified peptide epitope or mimotope.

30. The modified peptide epitope of any one of claims 10-29, wherein the unmodified peptide epitope or mimotope comprises or consists of no more than 10, no more than 15, nomore than 20, no more than 25, no more than 30, no more than 35, no more than 40, no more than 45, or no more than 50 consecutive amino acids in an autoimmune-associated antigen or a cancer-associated antigen.

31. A peptide epitope comprising or consisting of SEQ ID NO: 430, wherein the peptide epitope is capable of being bound by an anti-CCR4 antibody.

32. The peptide epitope of claim 31, wherein the anti-CCR4 antibody is mogamulizumab.

33. A peptide epitope comprising or consisting of any one of SEQ ID NO: 439-468, wherein the peptide epitope is capable of being bound by an anti-DLL3 antibody.

34. The peptide epitope of claim 33, wherein the anti-DLL3 antibody is tarlatamab.

35. A peptide epitope comprising or consisting of any one of SEQ ID NOs: 519-524, wherein the peptide epitope is capable of being bound by an anti-CCR7 antibody.

36. The peptide epitope of claim 35, wherein the anti-CCR7 antibody is CAP-100.

37. A recombinant polypeptide comprising or consisting of the peptide epitope or mimotope of any one of claims 10-36.

38. The recombinant polypeptide of claim 37, wherein the recombinant polypeptide consists of the peptide epitope or mimotope.

39. The recombinant polypeptide of claim 37 or 38, wherein the peptide epitope or mimotope is capable of being expressed on the surface of a cell.

40. The recombinant polypeptide of any one of claims 37-39, wherein the peptide epitope or mimotope is located in an extracellular region of the recombinant polypeptide.

41. The recombinant polypeptide of any one of claims 37-40, wherein the recombinant polypeptide is not any of CCR4, CCR7, DLL3, TROP2, GUCY2C, CD20, RSV protein F, EGFR, PD-1, CD34, and CD52.

42. The recombinant polypeptide of any one of claims 37-40, wherein the recombinant polypeptide is not CCR4.

43. The recombinant polypeptide of any one of claims 37-40, wherein the recombinant polypeptide is not CCR7.

44. The recombinant polypeptide of any one of claims 37-43, wherein the recombinant polypeptide does not comprise a cysteine at the C-terminus of the modified peptide epitope or mimotope.

45. The recombinant polypeptide of any one of claims 37-44, wherein the recombinant polypeptide does not comprise a cysteine immediately after the C-terminus of the modified peptide epitope or mimotope.

46. The recombinant polypeptide of any one of claims 37-45, wherein the recombinant polypeptide does not comprise a cysteine at the N-terminus of the modified peptide epitope or mimotope.

47. The recombinant polypeptide of any one of claims 37-46, wherein the recombinant polypeptide does not comprise a cysteine immediately before the N-terminus of the modified peptide epitope or mimotope.

48. The recombinant polypeptide of any one of claims 37-47, wherein the recombinant polypeptide comprises one or more additional peptide epitope(s) or mimotope(s), wherein the one or more additional peptide epitope(s) or mimotope(s) are the same as the modified peptide epitope or different from the modified peptide epitope.

49. A recombinant polypeptide comprising a tag, wherein the tag comprises or consists of a peptide epitope or a mimotope that mimics an epitope.

50. The recombinant polypeptide of claim 49, wherein the epitope is derived from CCR4, DLL3, or CCR7.

51. The recombinant polypeptide of any of claims 37-50, wherein the recombinant polypeptide comprises a binding region for an antigen.

52. The recombinant polypeptide of claim 51, wherein the antigen is expressed on the surface of a target cell.

53. The recombinant polypeptide of claim 51 or 52 wherein the recombinant polypeptide comprises or consists of a chimeric antigen receptor (CAR) or a recombinant T cell receptor (TCR).

54. The recombinant polypeptide of claim 51 or 52, wherein the recombinant polypeptide comprises or consists of a receptor comprising:(a) a heavy chain variable region (VH) located before the N-terminus of a constant region of TCRalpha, and a light chain variable region (VL) located before the N-terminus of a constant region of TCRbeta; or(b) a heavy chain variable region (VH) located before the N-terminus of a constant region of TCRbeta, and a light chain variable region (VL) located before the N-terminus of a constant region of TCRalpha, optionally, wherein at least one of the constant region of TCRalpha or the constant region of TCRbeta are an endogenous constant region of TCRalpha or an endogenous constant region of TCRbeta, optionally wherein the receptor is a HLA-independent T cell (HIT) receptor.

55. The recombinant polypeptide of any one of claims 51-54, wherein an immune cell expressing the recombinant polypeptide exhibits one or more of the following:(a) improved binding to the antigen expressed on the surface of the target cell;(b) reduced recombinant polypeptide-induced antigen-independent signaling and / or activation of the immune cell; and / or(c) reduced cross-linking between the recombinant polypeptides expressed on the cell surface of the immune cell; compared to an immune cell expressing a recombinant polypeptide comprising the unmodified peptide epitope or mimotope.

56. The recombinant polypeptide of any one of claims 51-55, wherein an immune cell expressing the recombinant polypeptide exhibits one or more of the following:(a) comparable levels of binding to the antigen expressed on the surface of the target cell;(b) comparable levels of recombinant polypeptide-induced antigen-independent signaling and / or activation of the immune cell; and / or(c) comparable levels of cross-linking between the recombinant polypeptides expressed on the cell surface of the immune cell; compared to an immune cell expressing a recombinant polypeptide that does not comprise a peptide epitope or mimotope.

57. The recombinant polypeptide of any one of claims 51-56, comprising the peptide epitope or mimotope located N-terminal to the binding region.

58. The recombinant polypeptide of claim 57, wherein the peptide epitope or mimotope is separated from the N-terminus of the binding region by no more than 15, no more than 10, no more than 5, no more than 4, no more than 3, no more than 2, no more than 1, or zero amino acid(s).

59. The recombinant polypeptide of any one of claims 51-58, comprising the peptide epitope or mimotope located C-terminal to the binding region.

60. The recombinant polypeptide of claim 59, wherein the peptide epitope or mimotope is separated from the C-terminus of the binding region by zero, no more than 5, no more than 10, no more than 15, no more than 20, no more than 25, no more than 30, no more than 35, no more than 40, no more than 45, no more than 50, no more than 55, or no more than 60 amino acids.

61. The recombinant polypeptide of any one of claims 51-60, wherein the peptide epitope or mimotope is separated from the binding region by no more than 10, no more than 15, no more than 20, no more than 25, no more than 30, no more than 35, no more than 40, no more than 45, or no more than 50 amino acids.

62. The recombinant polypeptide of any one of claims 51-61, wherein the peptide epitope or mimotope is located within the binding region.

63. The recombinant polypeptide of any one of claims 53-62, wherein the recombinant polypeptide comprises or consists of the CAR.

64. The recombinant polypeptide of any one of claims 51-63, wherein the binding region comprises a heavy chain variable domain (VH) and a light chain variable domain (VL).

65. The recombinant polypeptide of any one of claims 51-63, wherein the binding region comprises a single chain variable fragment (scFv) comprising a VH and a VL.

66. The recombinant polypeptide of any one of claims 51-63 , wherein the binding region comprises one or more variable heavy domains of heavy chain (VHHs).

67. The recombinant polypeptide of any one of claims 64-66, wherein the VH and the VL is separated by a linker, or wherein the binding region comprises at least two VHHs separated by a linker.

68. The recombinant polypeptide of any one of claims 51-63, wherein the binding region comprises a first binding domain and a second binding domain, wherein the first binding domain and the second binding domain are separated by a linker.

69. The recombinant polypeptide of any one of claims 51-63, wherein the recombinant polypeptide comprises a second binding region located N-terminal or C-terminal to the binding region; optionally, wherein the binding region and the second binding region are separated by a linker.

70. The recombinant polypeptide of any one of claims 67-69, wherein the peptide epitope or mimotope is located within the linker.

71. The recombinant polypeptide of any one of claims 67-70, wherein the linker comprises no more than 10, no more than 15, no more than 20, no more than 25, no more than 30, no more than 35, no more than 40, no more than 45, or no more than 50 amino acids.

72. The recombinant polypeptide of any one of claims 51-69, wherein the peptide epitope or mimotope is located in between the binding region(s) and a transmembrane region.

73. The recombinant polypeptide of any one of claims 51-69, wherein the peptide epitope or mimotope is located within a hinge region that connects the binding region(s) to a transmembrane region.

74. The recombinant polypeptide of claim 73, wherein the hinge region comprises no more than 10, no more than 15, no more than 20, no more than 25, no more than 30, no more than 35, no more than 40, no more than 45, no more than 50, no more than 55, no more than 60, no more than 65, or no more than 70 amino acids.

75. The recombinant polypeptide of any one of claims 53-74, wherein the CAR or TCR comprises no more than 10, no more than 15, no more than 20, no more than 25, no more than 30, no more than 35, no more than 40, no more than 45, or no more than 50 amino acids N-terminal to the binding region(s), excluding the signal peptide.

76. The recombinant polypeptide of any one of claims 53-75, wherein the extracellular region of the CAR or TCR comprises no more than 200, no more than 250, no more than 300, no more than 350, no more than 400, no more than 450, or no more than 500 amino acids.

77. The recombinant polypeptide of any one of claims 53-76, wherein the CAR or TCR comprises, from N-terminus to C-terminus, the binding region(s), a hinge region, a transmembrane region.

78. The recombinant polypeptide of any one of claims 51-77, wherein the antigen is an autoimmune-associated antigen or a cancer-associated antigen.

79. The recombinant polypeptide of any one of claims 37-78, wherein the peptide epitope is derived from an autoimmune-associated antigen or a cancer-associated antigen or wherein the mimotope mimics an epitope of the autoimmune-associated antigen or the cancer-associated antigen.

80. The recombinant polypeptide of claim 78 or 79, wherein the autoimmune-associated antigen or the cancer-associated antigen is selected from the group consisting of CD1, CD la, CDlb, CDlc, CDld, CDle, CD2, CD3d, CD3e, CD3g, CD3s, CD4, CD5, CD7, CD8a, CD8b, CD19, CD20, CD21, CD22, CD23, CD24, CD25, CD27, CD28, CD30, CD33, CD34, CD38, CD40, CD44v6, CD45, CD46, CD47 CD48, CD52, CD59, CD66, CD70, CD71, CD72, CD73, CD79A, CD79B, CD80 (B7.1), CD86 (B7.2), CD94, CD95, CD97, CD123, CD134, CD140 (PDGFR4), CD152, CD154, CD158, CD171, CD178, CD179, CD179a, CD181 (CXCR1), CD182 (CXCR2), CD183 (CXCR3), CD210, CD213A2, CD246, CD252, CD253, CD261, CD262, CD272, CD273 (PD-L2), CD274 (PD-L1), CD276 (B7H3), CD279, CD295, CD339 (JAG1), CD340 (HER2), CDH17, CEA, CLECL1, CLL-1, CLDN6, CLDN18.2, CS1, DLL3, LY6G6D, GCC, p53R175H, PRAME, EGFR, EGFRvIII, FGFR2, AFP, CA125, MUC-1, MAGE, ALPI, alkaline phosphatase placental- like 2 (ALPPL2), B-cell maturation antigen (BCMA), green fluorescent protein (GFP), enhanced green fluorescent protein (eGFP), KLK2, KLK3, Mesothelin, IL13Ra2, signal regulatory protein a (SIRPa), TCRalpha, TCRbeta, TSHR, GD2, GD3, Tn Ag, cMET, Axl, ROR1, ROR2, GPC1, GPC2, GPC3, FLT3, TAG72, CEA, EPCAM, KIT (CD117), IL- 13Ra2, IL-l lRa, PSCA, PRSS21, VEGFR2, LewisY, PDGFRp, SSEA-4, folate receptor alpha, ERBB2 (Her2 / neu), MUC1, MUC16, NCAM, prostase, PAP, ELF2M, Ephrin B2, IGF -I receptor, CAIX, LMP2, gplOO, bcr-abl, tyrosinase, EphA2, STEAP1, STEAP2, fucosyl GM1, sLe, GM3, TGS5, HMWMAA, o-acetyl-GD2, folate receptor beta, TEM1 / CD248, TEM7R, ROPN1, GPRC5D, GPA33, CXORF61, ALK, Polysialic acid,PLAC1, GloboH, NY-BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP, WT1, NY-ESO-1, LAGE-la, MAGE-A1, legumain, HPV E6,E7, MAGE-A4, ETV6- AML, sperm protein 17, XAGE1, Tie 2, MAD-CT-1, MAD-CT-2, Fos-related antigen 1, p53, p53 mutant, p53R175H, KRAS, mutant KRAS, KRAS G12D, prostein, surviving, telomerase, PCTA-l / Galectin 8, MelanA / MARTl, Ras mutant, hTERT, sarcoma translocation breakpoints, ML-IAP, ERG (TMPRSS2 ETS fusion gene), NA17, PAX3, androgen receptor, cyclin Bl, MYCN, RhoC, TRP-2, CYP1B1, BORIS, SART3, PAX5, OY-TES1, LCK, AKAP-4, SSX2, RAGE-1, human telomerase reverse transcriptase, RU1, RU2, intestinal carboxyl esterase, mut hsp70-2,LAIRl, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, FCRL5, IGLL1, PSMA, TROP2, citrullinated vimentin, the extracellular portion of the APRIL protein, and any combinations thereof.

81. The recombinant polypeptide of any one of claims 78-80, wherein the cancer- associated antigen is a cancer-associated antigen for a solid tumor.

82. The recombinant polypeptide of claim 81, wherein the cancer-associated antigen for the solid tumor comprises or consists of KIR2D, NRP1, CD221, VEGFR, HGF, integrin a5pi, CD51, CD27, CDH17, B7H3, NYESO1, EGFR, cMET, BRD4, EpCAM, SSX, Fibronectin EDB, integrin al ipi, DLL3, CD2, SSTR2, TROP2, KLK2, KLK3, STEAP2, STEAP1, PSMA, PSCA, LY6G6D, IL- la, CEA, C242 antigen, CanAg (MUC1 glycoform), CTAA16.88, UCY2C, IGF2BP3, p53R175H, NKG2D ligands, PRAME, MAGE-A4, vimentin, HER2 / neu, CCR4, CCR5, ROPN1, GPA33, ROR1, KK-LC-1, CA-125, folate receptor 1, folate receptor alpha, MUC1, MUC16, MSLN, CLDN6, WT1, ALPPL2, KRASG12D, KRASG12V, CD70, MSLN, CLDN18.2, HPV E6 / E7, GPC3, IL-13Ra, EGFRv3, EGFR806, GD2, or CA9.

83. The recombinant polypeptide of claim 82, wherein the cancer-associated antigen for the solid tumor comprises or consists of KIR2D, NRP1, CD221, VEGFR, HGF, integrin a5bl, CD51, CD27, B7H3, NYESO1, EGFR, cMET, BRIM, EpCAM, SSX, Fibronectin EDB, or Integrin al ipi.

84. The recombinant polypeptide of claim 82, wherein the cancer-associated antigen for the solid tumor comprises or consists of KLK2, Ly6G6D, CLDN6, DLL3 or CD 19.

85. The recombinant polypeptide of any one of claims 78-84, wherein the cancer is a solid tumor selected from the group consisting of small cell lung cancer, prostate cancer, colorectal cancer, head and neck squamous cell carcinoma, breast cancer, melanoma, ovarian cancer, lung cancer, pancreatic cancer, stomach cancer, cervical cancer, hepatocellular carcinoma, glioma cancer, neuroblastoma cancer, and renal cell carcinoma.

86. The recombinant polypeptide of any one of claims 78-85, wherein the cancer is a hematologic cancer selected from B cell lymphoma, T cell lymphoma, chronic lymphocytic leukemia, and acute myeloid leukemia.

87. The recombinant polypeptide of any one of claims 78-86, wherein the cancer- associated antigen is or comprises an antigen selected from Table 2.

88. The recombinant polypeptide of any one of claims 51-87, wherein the antigen is DLL3.

89. The recombinant polypeptide of any one of claims 37-88, wherein the epitope is derived from a cancer-associated antigen selected from Table 2.

90. The recombinant polypeptide of any one of claims 37-89, wherein the epitope is derived from DLL3.

91. The recombinant polypeptide of claim 90, wherein the peptide epitope comprises or consists of at least 10, at least 15, at least 20, at least 25, at least 30, at least 35, or at least 40 consecutive amino acids in SEQ ID NO: 486, or an amino acid sequence having at most 1, at most 2, at most 3, at most 4, or at most 5 amino acid mutations thereto.

92. The recombinant polypeptide of claim 90, wherein the peptide epitope comprises or consists of any one of SEQ ID NO: 439-468, or an amino acid sequence having at most 1, at most 2, at most 3, at most 4, or at most 5 amino acid mutations thereto.

93. The recombinant polypeptide of claim 90, wherein the peptide epitope comprises or consists of SEQ ID NO: 439, or an amino acid sequence having at most 1, at most 2, at most 3, at most 4, or at most 5 amino acid mutations thereto.

94. The recombinant polypeptide of any one of claims 51-93, wherein the peptide epitope or mimotope is capable of being bound by a monoclonal antibody that binds to the antigen for the binding region.

95. The recombinant polypeptide of any one of claims 37-87, wherein the epitope is derived from a cancer-associated antigen of T cell lymphoma.

96. The recombinant polypeptide of any one of claims 37-87, wherein the epitope is derived from a cancer-associated antigen of adult T-cell lymphoma (ATL) and / or cutaneous T-cell lymphoma (CTCL).

97. The recombinant polypeptide of any one of claims 37-87, 95, and 96, wherein the epitope is derived from a cancer-associated antigen of a Th2 cell, a cutaneous lymphocyte antigen-positive skin-homing T cell, and / or a Treg cell.

98. The recombinant polypeptide of any one of claims 37-87 and 95-97, wherein the epitope is derived from CCR4.

99. The recombinant polypeptide of claim 98, wherein the peptide epitope comprises or consists of an amino acid sequence of any one of SEQ ID NO: 430-438, or an amino acid sequence having at most 1, at most 2, at most 3, at most 4, or at most 5 amino acid mutations thereto.

100. The recombinant polypeptide of claim 98, wherein the peptide epitope comprises or consists of an amino acid sequence of SEQ ID NO: 431.

101. The recombinant polypeptide of any one of claims 37-87 and 95-97, wherein the epitope is derived from CCR7.

102. The recombinant polypeptide of claim 101, wherein the peptide epitope comprises or consists of an amino acid sequence of any one of SEQ ID NOs: 519-524, or an amino acid sequence having at most 1, at most 2, at most 3, at most 4, or at most 5 amino acid mutations thereto.

103. The recombinant polypeptide of claim 101, wherein the peptide epitope comprises or consists of an amino acid sequence of SEQ ID NOs: 519-524.

104. The recombinant polypeptide of any one of claims 37-103, wherein the peptide epitope or the mimotope is about 5 to about 10, about 10 to about 20, about 15 to about 25, about 20 to about 30, about 25 to about 35, about 30 to about 40, about 35 to about 45, or about 40 to about 50 amino acids in length.

105. The recombinant polypeptide of any one of claims 37-104, wherein the peptide epitope or the mimotope is no more than 10, no more than 15, no more than 20, no more than 25, no more than 30, no more than 35, no more than 40, no more than 45, or no more than 50 amino acids in length.

106. The recombinant polypeptide of any one of claims 37-105, wherein the peptide epitope does not comprise any mutation compared to the native epitope derived from the antigen.

107. The recombinant polypeptide of any one of claims 37-106, wherein the peptide epitope or the mimotope is capable of forming an alpha-helical conformation.

108. The recombinant polypeptide of any one of claims 37-107, wherein the peptide epitope or the mimotope is capable of being bound by an antibody with a dissociation constant (Kd) of less than 1 pM, less than 100 nM, less than 10 nM, or less than 1 nM, optionally wherein the Kd is measured by surface plasmon resonance (SPR) method using a sensor chip that contains the immobilized antibody.

109. The recombinant polypeptide of claim 108, wherein the antibody is [fam- ]trastuzumab deruxtecan, (fam-trastuzumab deruxtecan-nxki), Adebrelimab, Ado-trastuzumab emtansine, Alemtuzumab, Amivantamab, Atezolizumab, Avelumab, Belantamab mafodotin (belantamab mafodotin-blmf), Bevacizumab, Blinatumomab, Brentuximab vedotin, Cadonilimab, Camrelizumab, CAP- 100, Catumaxomab, Cemiplimab (cemiplimab-rwlc), Cetuximab, Cetuximab saratolacan, Cosibelimab, Daratumumab, Dinutuximab, Disitamab vedotin, Dostarlimab, Durvalumab, Edrecolomab, Elotuzumab, Elranatamab, Emapalumab (emapalumab-lzsg), Enfortumab vedotin (enfortumab vedotin- ejfv), Enlonstobart, Envafolimab, Epcoritamab, Gemtuzumab ozogamicin, Glofitamab, Ibritumomab tiuxetan, Inotuzumab ozogamicin, Ipilimumab, Isatuximab (isatuximab-irfc), Loncastuximab tesirine, Margetuximab-cmkb, Mirvetuximab soravtansine, Mogamulizumab (mogamulizumab-kpkc), Mosunetuzumab, Moxetumomab pasudotox (moxetumomab pasudotox-tdfk), Naxitamab-gqgk, Necitumumab, Nimotuzumab, Nivolumab, Cipterbin, Obinutuzumab, Ofatumumab, Olaratumab, Panitumumab, Pembrolizumab, Penpulimab, Pertuzumab, Polatuzumab vedotin (polatuzumab vedotin-piiq), Prolgolimab, Pucotenlimab, Racotumomab, Ramucirumab, Relatlimab, Retifanlimab, Ripertamab, Rituximab, Sacituzumab govitecan (sacituzumab govitecan-hziy), Serplulimab, Sintilimab, Socazolimab, Sugemalimab, Tafasitamab (tafasitamab-cxix), Tagitanlimab, Talquetamab (talquetamab-tgvs), Tebentafusp, Teclistamab, Tislelizumab, Tisotumab vedotin, tisotumab vedotin-tftv, Toripalimab, Tositumomab-1131, Trastuzumab, Trastuzumab duocarmazine, Tremelimumab, Zimberelimab, Zolbetuximab, Zuberitamab, Odronextamab, Ivonescimab, Benmel Stobart, Trastuzumab botidotin, Iparomlimab, tuvonralimab, Tarlatamab, Patritumab deruxtecan, Sacituzumab tirumotecan, Zanidatamab, Linvoseltamab, Datopotamab deruxtecan, Anbenitamab, Anvatabart opadotin, Apamistamab-Iodine (1311), Bemarituzumab, Cetrelimab, Cobolimab, Domvanalimab, Emactuzumab, Erfonrilimab, Favezelimab, Felzartamab, Fianlimab, Finotonlimab, ABBV-383, Geptanolimab, Gotistobart, Ivuxolimab, Izalontamab, Lemzoparlimab, Luveltamab tazevibulin, Magrolimab, Mecbotamab vedotin, Monalizumab, Nofazinlimab, Nurulimab, Ociperlimab, Oleclumab, Onfekafusp alfa, Oregovomab, Ozekibart, Pivekimab sunirine, Quavonlimab, Retlirafusp alfa, Rosopatamab (177Lu-DOTA), Rulonilimab, Sasanlimab, Telisotuzumab vedotin, Tiragolumab, IKS014, LCB14-0110, FS-1502, Trastuzumab mafodotin, Trastuzumab rezetecan, Vibostolimab, Vobramitamab duocarmazine, Zilovertamab vedotin, Suvemcitug, APX003, BD0801, TK001, sevacizumab, CTX-009, ESI 04, TR009, NOV1501, ABL001, DP303c, IAH0968, Kintuximab, gentuximab, cintuximab, MIL62, MRG002, MRG003, PM8002, MEDI5752, MCLA-128, AGEN1181, L19-IL2, L19-TNF,IMC-F106C, JMT101, BL-B01D1, BAT13O8, AZD2936, JS004, TAB004, FG-M108, M1O8, 9MW2821, BMS-986349, CC-93269, EM801, ABBV-181, ABBV-151, ARGX-115, DS-7300a, IBI343, SHR-A1921, or ASKB589 .

110. The recombinant polypeptide of claim 108, wherein the antibody is tarlatamab.

111. The recombinant polypeptide of claim 108, wherein the antibody is mogamulizumab.

112. The recombinant polypeptide of claim 108, wherein the antibody is CAP-100.

113. The recombinant polypeptide of any one of claims 37-112, wherein the recombinant polypeptide comprises one or more additional peptide epitope(s) or mimotope(s).

114. The recombinant polypeptide of claim 113, wherein the one or more additional peptide epitope(s) or mimotope(s) are the same as the peptide epitope or mimotope in the tag.

115. The recombinant polypeptide of claim 113, wherein the one or more additional peptide epitope(s) or mimotope(s) are different from the peptide epitope or mimotope in the tag.

116. The recombinant polypeptide of any one of claims 113-115, wherein the recombinant polypeptide comprises 1, 2, 3, or 4 additional peptide epitope(s) or mimotope(s).

117. The recombinant polypeptide of any one of claims 37-116, wherein the recombinant polypeptide comprises an extracellular portion comprising the binding region and the peptide epitope or mimotope.

118. The recombinant polypeptide of any one of claims 37-117, wherein the recombinant polypeptide comprises a transmembrane region or a glycosylphosphatidylinositol (GPI) anchor.

119. The recombinant polypeptide of any one of claims 37-118, wherein the recombinant polypeptide comprises an intracellular region; optionally, the intracellular domain is located C-terminal to the transmembrane region.

120. The recombinant polypeptide of claim 119, wherein the intracellular region comprises one or more intracellular signaling domains.

121. A recombinant nucleic acid encoding the peptide epitope or mimotope of any one of claims 10-36, or the recombinant polypeptide of any one of claims 37-120.

122. A recombinant nucleic acid encoding (i) a recombinant receptor that comprises a binding region for an antigen, and (ii) a recombinant polypeptide comprising a tag in its extracellular region, wherein the tag comprises a peptide epitope or a mimotope that mimics an epitope.

123. The recombinant nucleic acid of claim 121 or 122, wherein the recombinant nucleic acid further comprises a polynucleotide sequence encoding one or more additional polypeptides and / or a non-coding RNA, optionally wherein the non-coding RNA comprises a shRNA or a microRNA.

124. The recombinant nucleic acid of claim 123, wherein the one or more additional polypeptides comprise an additional potency enhancement polypeptide, a cytokine, a chemokine, a growth factor, or any combination thereof.

125. The recombinant nucleic acid of any one of claims 121-124, wherein the recombinant nucleic acid is less than 10 kb, less than 9 kb, less than 8 kb, less than 7 kb, less than 6 kb, less than 5 kb, less than 4 kb, or less than 3 kb in length.

126. The recombinant nucleic acid of claim 125, wherein the recombinant nucleic acid is less than about 5.0 kb in length.

127. The recombinant nucleic acid of claim 126, wherein the recombinant nucleic acid is less than about 4.7 kb in length.

128. A vector comprising the recombinant nucleic acid of any one of claims 121-127.

129. The vector of claim 128, wherein the vector is a viral vector.

130. The vector of claim 129, wherein the viral vector is selected from a retrovirus vector, an adenovirus vector, and an adeno-associated virus (AAV) vector; optionally, wherein the viral vector is the AAV vector.

131. An engineered cell, wherein the engineered cell (i) comprises the recombinant nucleic acid of any one of claims 121-127, or (ii) is transfected or transduced by the vector of any one of claims 128-130.

132. An engineered cell expressing the peptide epitope or mimotope of any one of claims 10-36, or the recombinant polypeptide of any one of claims 37-120.

133. An engineered cell expressing (i) a recombinant receptor that comprises a binding region for an antigen, and (ii) a recombinant polypeptide comprising a tag in its extracellular region, wherein the tag comprises a peptide epitope or a mimotope that mimics an epitope.

134. An engineered cell comprising the recombinant polypeptide of any one of claims 37- 120 or the recombinant nucleic acid of any one of claims 121-127.

135. The engineered cell of any one of claims 131-134, wherein the engineered cell comprises one or more additional peptide epitope(s) or mimotope(s) present on the cell surface, wherein the one or more additional peptide epitope(s) or mimotope(s) are the same as the peptide epitope or mimotope or different from the peptide epitope or mimotope.

136. The engineered cell of any one of claims 131-135, wherein the engineered cell exhibits one or more of the following:(a) improved binding to the antigen expressed on the surface of a target cell;(b) reduced recombinant polypeptide-induced antigen-independent signaling and / or activation of the engineered cell; and / or(c) reduced cross-linking between the recombinant polypeptides expressed on the cell surface of the engineered cell; compared to a control cell expressing a recombinant polypeptide comprising the unmodified peptide epitope or mimotope.

137. The engineered cell of any one of claims 131-136, wherein the engineered cell exhibits one or more of the following:(a) comparable levels of binding to the antigen expressed on the surface of a target cell;(b) comparable levels of recombinant polypeptide-induced antigen-independent signaling and / or activation of the engineered cell; and / or(c) comparable levels of cross-linking between the recombinant polypeptides expressed on the cell surface of the engineered cell; compared to a control cell expressing a recombinant polypeptide that does not comprise the peptide epitope or mimotope.

138. A composition comprising the recombinant nucleic acid of any one of claims 121- 127, the vector of any one of claims 128-130, or the engineered cell of any one of claims 131-137.

139. A method of preparing an engineered cell, wherein the method comprises contacting the cell with the recombinant nucleic acid of any one of claims 121-127, or the vector of any one of claims 128-130.

140. A method of preparing an engineered cell, wherein the method comprises expressing in the cell:(a) the recombinant polypeptide of any one of claims 37-120; or(b) at least the following two polypeptides: (i) a recombinant receptor that comprises a binding region for an antigen, and (ii) a recombinant polypeptide comprising at least one tag in its extracellular region, wherein the at least one tag comprises a peptide epitope or a mimotope that mimics an epitope.

141. The method of claim 139 or 140, wherein the step of introducing comprises homology-directed repair (HDR)-mediated insertion using the gene editing machinery.

142. The method of claim 139, wherein the recombinant nucleic acid or the vector is introduced via AAV.

143. The method of any one of claims 139-142, wherein the method thereby produces an engineered cell comprising: (i) the recombinant polypeptide; (ii) the immune receptor (e.g., CAR); and (iii) the modified TRAC gene locus.

144. The method of any one of claims 139-142, wherein the method thereby produces an engineered cell comprising: (i) the recombinant polypeptide; (ii) the immune receptor (e.g., CAR); and (iii) the modified TRAJ locus.

145. The method of any one of claims 139-144, wherein the cell is for use in a cell therapy.

146. A method of treating cancer or an autoimmune disease in a subject in need thereof, comprising administering an effective amount of the engineered cells of any one of claims 131-137 to the subject.

147. A method of treating an autoimmune disease in a subject in need thereof, comprising administering an effective amount of the recombinant nucleic acid of any one of claims 121- 127, or the vector of any one of claims 128-130, to the subject.

148. A method of treating cancer in a subject in need thereof, comprising administering an effective amount of the recombinant nucleic acid of any one of claims 121-127, or the vector of any one of claims 128-130, to the subject.

149. The method of any one of claims 146-148, wherein the method further comprises administering an antibody to the subject, wherein the antibody binds the tag, the peptide epitope, or the mimotope.

150. The method of any one of claims 146-149, wherein the method further comprises monitoring the subject for an adverse event.

151. The method of any one of claims 146-150, wherein the method further comprises monitoring the subject for treatment outcomes.

152. A method of depleting engineered cells in a subject in need thereof, comprising administering to the subject an antibody that binds to a tag expressed on the surface of theengineered cells, wherein the tag comprises a peptide epitope or a mimotope that mimics an epitope.

153. A method of reducing or eliminating at least one symptom of an adverse event caused by engineered cells in a subject in need thereof, comprising administering to the subject an antibody that binds to a tag expressed on the surface of the engineered cells, wherein the tag comprises a peptide epitope or a mimotope that mimics an epitope.

154. The method of any one of claims 149-153, wherein administration of the antibody depletes at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 99%, at least 99.5%, or at least 99.9% of the engineered cells expressing the tag, the peptide epitope, or the mimotope in the subject.

155. The method of any one of claims 149-154, wherein the antibody is administered after the detection or occurrence of an adverse event, and wherein administration of the antibody reduces or eliminates at least one symptom of the adverse event.

156. The method of any one of claims 149-155, wherein the antibody is tarlatamab.

157. The method of any one of claims 149-155, wherein the antibody is mogamulizumab.

158. The method of any one of claims 149-155, wherein the antibody is CAP-100.

159. A method of isolating or enriching the engineered cells of any one of claims 131-137, comprising contacting the engineered cells with a binding agent that binds to the epitope or the mimotope.

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