Methods for treating cancer

Abstract

The disclosure describes scFv- and nanobody-based CARs against MSLN and their use in treating cancer. Also described are engineered CAR-T cells that can simultaneously target a surface-bound tumor antigen and secrete immunostimulatory cytokines and chemokines can not only enable direct tumor killing but also recruit endogenous immunity against the tumor. The synergy of multiple cytokines was examined, and it was found that IL-12 plus an IL-18 variant (DR-18) significantly increased CAR-T cell efficacy against GBM and melanoma. A further combination includes IL-12, IFN-β, and CXCL9, which also showed increased CAR-T cell efficacy against cancer cells.

Description

METHODS FOR TREATING CANCER

[0001] This application claims priority of U.S. Provisional Application No. 63 / 536,856, filed September 6, 2023; U.S. Provisional Application No.63 / 551,372, filed February 8, 2024; U.S. Provisional Application No. 63 / 536,862, filed September 6, 2023; U.S. Provisional Application No. 63 / 551,449, filed February 8, 2024; U.S. Provisional Application No. 63 / 536,868, filed September 6, 2023; and U.S. Provisional Application No.63 / 551,398, filed February 8, 2024, which are hereby incorporated by reference in their entirety.

[0002] The application contains a Sequence Listing in compliance with ST.26 format and is hereby incorporated by reference in its entirety. Said Sequence Listing, created on September 4, 2024 is named UCLAP0188WO.XML and is 323,142 bytes in size. BACKGROUND I. Field of the Invention

[0003] This invention relates to the field of immunology and cancer therapeutics. II. Background

[0004] Chimeric antigen receptor (CAR)-expressing T cells have become an important immunotherapy, demonstrating remarkable clinical efficacy against B-cell malignancies. All of the clinically approved CAR-T cell products on the current market utilize single-chain variable fragments (scFvs) as the extracellular binding domain which confers target specificity and ability of CAR-T cells to recognize and target antigen-expressing tumor cells. These scFvs are fusion proteins originating from the variable heavy and light chains of antibodies connected by a short peptide linker. The specificity of scFvs has made it an attractive modality to redirect T-cell specificity towards tumor cells. scFvs are typically 240–250 amino acids in length, often accounting for the majority of the genetic footprint of the CAR transgene. A reduction in the size of the ligand-binding domain could increase the transduction efficiency and / or enable the incorporation of additional transgenic elements the further increase the efficacy of CAR-T cell therapy. In addition, reducing the size of the ligand-binding domain could also increase stability and reduce immunogenicity of the CAR construct. Furthermore, many scFvs, antibodies, and other extracellular binding modalities used in CARs demonstrate reactivity only against the antigen from one species (e.g., human or mouse). This hampers clinical translation as CARs that only recognize human antigens cannot be used in preclinical immunocompetent murine202666783.1 - 1 -models and vice versa. This, in turn, prevents the rigorous assessment of the CAR-T cells’ ability to interact with endogenous immunity and the tumor microenvironment.

[0005] Mesothelin (MSLN) is an antigen in which scFvbased CARs have been developed for and have been tested in the clinic against cancers such as ovarian cancer, pancreatic cancer, lung cancer, and mesothelioma. There are no known MSLN CARs that have been reported to date do not cross-recognize murine MSLN. SUMMARY

[0006] The disclosure provides for a chimeric polypeptide comprising: (a) an antigen binding domain comprising a nanobody, wherein the nanobody comprises a complementarity determining region (CDR) 1, CDR2, and CDR3 having at least 80% sequence identity to the CDR1, CDR2, CDR3 from the nanobody of SEQ ID NO:57; (b) a transmembrane domain; and (c) an intracellular signaling domain. Also described is a chimeric polypeptide comprising: (a) an antigen binding domain comprising a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:36 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:37; (b) a transmembrane domain; and (c) an intracellular signaling domain. Also described is a chimeric polypeptide comprising: (a) an antigen binding domain comprising a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:45 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:46; (b) a transmembrane domain; and (c) an intracellular signaling domain. Also provided is a chimeric polypeptide comprising: (a) an antigen binding domain comprising a nanobody, wherein the nanobody comprises a complementarity determining region (CDR) 1, CDR2, and CDR3 having at least 80% sequence identity to the CDR1, CDR2,202666783.1 - 2 -CDR3 from the nanobody of SEQ ID NO:53; (b) a transmembrane domain; and (c) an intracellular signaling domain.

[0007] The disclosure provides for an anti-VEGF antibody or antigen binding fragment comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:2 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:3. Also provided is an anti-VEGF antibody or antigen binding fragment comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:19 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:20. Also included are polypeptides comprising the antigen binding fragment of the disclosure and polypeptides expressed from the nucleic acids of the disclosure as well as cells or population of cells comprising the nucleic acids or polypeptides of the disclosure. Further provided are cells or populations of cells that express the nucleic acid(s) of the disclosure. Also described herein is a chimeric polypeptide comprising: (a) an antigen binding fragment of the disclosure; (b) a transmembrane domain; and (c) an intracellular signaling domain. Included are one or more nucleic acids that encode the antibody, antigen binding fragment, and / or polypeptide of the disclosure. Included are compositions comprising the cells, nucleic acids, or polypeptides of the disclosure. The composition may be a pharmaceutically acceptable formulation.

[0008] Also described is a polypeptide comprising the amino acid sequence of any one of SEQ ID NOS:138-207 or an amino acid sequence having at least 80% sequence identity to the amino acid sequences of SEQ ID NOS:138-207. The polypeptide may have or have at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the amino acid sequence of any one of SEQ ID NOS:138-207. Also provided is a nucleic acid encoding a polypeptide comprising the amino acid sequence of one of SEQ ID NOS:138-207 or an amino acid sequence having at least 80% sequence identity to one of202666783.1 - 3 -SEQ ID NOS:138-207. The nucleic acid may encode a polypeptide that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the amino acid sequence of any one of SEQ ID NOS:138-207.

[0009] Also described is a nucleic acid encoding: (i) a chimeric antigen receptor (CAR) or engineered T-cell receptor (TCR); and (ii) an IL-12 polypeptide; and (iii) an IL-18 polypeptide. Also described is a nucleic acid encoding: (i) a chimeric antigen receptor (CAR) or engineered T-cell receptor (TCR); and (ii) an IL-12 polypeptide; (iii) an IFN-β polypeptide; and (iv) a CXCL9 polypeptide. Also provided are cells or a population of cells comprising a heterologous nucleic acid encoding an IL-12 polypeptide and a heterologous nucleic acid encoding an IL-18 polypeptide. Cells also include those that comprise a heterologous nucleic acid encoding an IL-12 polypeptide, a heterologous nucleic acid encoding an IFN-β polypeptide and a heterologous nucleic acid encoding a CXCL9 polypeptide.

[0010] Also provided are nucleic acid(s) comprising a sequence encoding a polypeptide of the disclosure, nucleic acid(s) comprising a sequence encoding an antibody or antigen binding fragment of the disclosure, a lentivirus vector comprising a sequence encoding a chimeric polypeptide of the disclosure, a polypeptide expressed from the nucleic acids of the disclosure, a cell or population of cells comprising a nucleic acid of the disclosure, and a cell or population of cells expressing the nucleic acid of the disclosure. Also described are compositions comprising cells, polypeptides, CARs, or nucleic acids of the disclosure. The composition may be a pharmaceutically acceptable formulation.

[0011] Methods include a method of making a cell comprising introducing into a cell a nucleic acid of the disclosure. Methods include a method of treating a patient with cancer comprising administering to the patient an effective amount of a composition of the disclosure. Also described is a method for increasing anti-tumor immunity and / or tumor cell cytotoxicity in a patient having cancer, the method comprising administering to the patient an effective amount of a composition of the disclosure.

[0012] The nucleic acids of the disclosure may encode for a chimeric antigen receptor (CAR), wherein the CAR comprises: (a) an antigen binding domain (b) a transmembrane domain; and (c) an intracellular signaling domain.

[0013] The antigen binding domain may comprise anti-MSLN. The antigen binding domain may comprise a nanobody, wherein the nanobody comprises a complementarity determining region (CDR) 1, CDR2, and CDR3 having at least 80% sequence identity to the CDR1, CDR2, CDR3 from the nanobody of SEQ ID NO:57. The nanobody may comprise a202666783.1 - 4 -CDR1, CDR2, and CDR3 having an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the CDR1, CDR2, CDR3 from the nanobody of SEQ ID NO:57. The nanobody may comprise a CDR1, CDR2, and CDR3 having the amino acid sequence of the CDR1, CDR2, and CDR3, respectively, from the nanobody of SEQ ID NO:57. The CDR1, CDR2, and CDR3 may each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:58-60, respectively. The CDR1, CDR2, and CDR3 may each comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NOS:58-60, respectively. The CDR1, CDR2, and CDR3 may each comprise the amino acid sequence of SEQ ID NOS:58-60, respectively. The nanobody may comprise a framework region (FR) 1, FR2, FR3, and FR4, wherein the FR1, FR2, FR3, and FR4 comprises an amino acid sequence with at least 80% sequence identity to a FR1, FR2, FR3, and FR4, respectively, of a nanobody comprising the amino acid sequence of SEQ ID NO:57. The nanobody may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to a FR1, FR2, FR3, and FR4, respectively, of a nanobody comprising the amino acid sequence of SEQ ID NO:57. The FR1, FR2, FR3, and FR4 may comprise the amino acid sequence of SEQ ID NOS:225-228, respectively or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:225-228, respectively. The FR1, FR2, FR3, and FR4 may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the amino acid sequence of SEQ ID NOS:225-228, respectively.

[0014] The antigen binding domain may comprise a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:36 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID202666783.1 - 5 -NO:37. The VH may comprise a HCDR1, HCDR2, and HCDR3 comprising an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:36. The light chain variable region may comprise a LCDR1, LCDR2, and LCDR3 comprising an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:37. The VH may comprise a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:36. The the VL may comprise a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:37. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:38-43, respectively. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NOS:38-43, respectively. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise the amino acid sequence of SEQ ID NOS:38-43, respectively. The VH may comprise an amino acid sequence with at least 70% sequence identity to SEQ ID NO:36 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:37. The VH may comprise an amino acid that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:36. The VL may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:37. The VH may comprise the amino acid sequence of SEQ ID NO:36. The VL may comprise the amino acid sequence of SEQ ID NO:37. The VH may comprise a heavy chain framework region (HFR) 1, HFR2, HFR3, and HFR4 and the VL comprises a light chain framework region (LFR) 1, LFR2, LFR3, and LFR4, and wherein the HFR1, HFR2, HFR3, and HFR4 may comprise an amino202666783.1 - 6 -acid sequence with at least 80% sequence identity to an HFR1, HFR2, HFR3, and HFR4, respectively, of a VH comprising the amino acid sequence of SEQ ID NO:36, and the LFR1, LFR2, LFR3, and LFR4 may comprise an amino acid sequence with at least 80% sequence identity to the LFR1, LFR2, LFR3, and LFR4, respectively, of a VL comprising the amino acid sequence of SEQ ID NO:37. The HFR1, HFR2, HFR3, and HFR4 may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to an HFR1, HFR2, HFR3, and HFR4, respectively, of a VH comprising the amino acid sequence of SEQ ID NO:36. The LFR1, LFR2, LFR3, and LFR4 may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the LFR1, LFR2, LFR3, and LFR4, respectively, of a VL comprising the amino acid sequence of SEQ ID NO:37. The HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4 may comprise the amino acid sequence of SEQ ID NOS:208-215, respectively, or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:208-215, respectively. The HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4 may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the amino acid sequence of SEQ ID NOS:208-215, respectively.

[0015] The antigen binding domain may comprise a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:45 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:46. The VH may comprise a HCDR1, HCDR2, and HCDR3 comprising an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:45. The light chain variable region may comprise a LCDR1, LCDR2, and LCDR3 comprising an amino acid sequence that has or has202666783.1 - 7 -at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:46. The VH may comprise a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:45. The VL may comprise a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:46. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:47-52, respectively. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NOS:47-52, respectively. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise the amino acid sequence of SEQ ID NOS:47-52, respectively. The VH may comprise an amino acid sequence with at least 70% sequence identity to SEQ ID NO:45 and / or the VL may comprise an amino acid sequence with at least 70% sequence identity to SEQ ID NO:46. The VH may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:45. The VL may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:46. The VH may comprise the amino acid sequence of SEQ ID NO:45. The VL may comprise the amino acid sequence of SEQ ID NO:46. The VH may comprise a heavy chain framework region (HFR) 1, HFR2, HFR3, and HFR4 and the VL may comprise a light chain framework region (LFR) 1, LFR2, LFR3, and LFR4, and wherein the HFR1, HFR2, HFR3, and HFR4 may comprise an amino acid sequence with at least 80% sequence identity to an HFR1, HFR2, HFR3, and HFR4, respectively, of a VH comprising the amino acid sequence of SEQ ID NO:45, and the LFR1, LFR2, LFR3, and LFR4 may comprise an amino acid sequence with at least 80% sequence identity to the LFR1, LFR2, LFR3, and LFR4, respectively, of a VL comprising the amino acid sequence of SEQ ID NO:46. The HFR1, HFR2, HFR3, and HFR4 may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66,202666783.1 - 8 -67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to an HFR1, HFR2, HFR3, and HFR4, respectively, of a VH comprising the amino acid sequence of SEQ ID NO:45. The LFR1, LFR2, LFR3, and LFR4 may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the LFR1, LFR2, LFR3, and LFR4, respectively, of a VL comprising the amino acid sequence of SEQ ID NO:46. The HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4 may comprise the amino acid sequence of SEQ ID NOS:216, 209, 217, 211, 218, 219, 220, and 215, respectively or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:216, 209, 217, 211, 218, 219, 220, and 215, respectively. The HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4 may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the amino acid sequence of SEQ ID NOS:216, 209, 217, 211, 218, 219, 220, and 215, respectively.

[0016] The antigen binding domain may comprise a nanobody. The nanobody may comprise a complementarity determining region (CDR) 1, CDR2, and CDR3 having at least 80% sequence identity to the CDR1, CDR2, CDR3 from the nanobody of SEQ ID NO:53. The nanobody may comprise a CDR1, CDR2, and CDR3 comprising an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the CDR1, CDR2, CDR3 from the nanobody of SEQ ID NO:53. The nanobody may comprise a CDR1, CDR2, and CDR3 having the amino acid sequence of the CDR1, CDR2, and CDR3, respectively, from the nanobody of SEQ ID NO:53. The CDR1, CDR2, and CDR3 may each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:54-56, respectively. The CDR1, CDR2, and CDR3 may each comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NOS:54-56, respectively. The CDR1, CDR2, HCDR2, LCDR1, LCDR2, and CDR3 may each comprise the amino acid sequence of SEQ ID NOS:54-56, respectively. The nanobody may comprise a framework region (FR) 1, FR2, FR3, and FR4, wherein the FR1, FR2, FR3, and FR4 comprises an amino acid sequence with at least 80% sequence identity to202666783.1 - 9 -a FR1, FR2, FR3, and FR4, respectively, of a nanobody comprising the amino acid sequence of SEQ ID NO:53. The nanobody may comprise a FR1, FR2, FR3, and FR4, wherein the FR1, FR2, FR3, and FR4 may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to a FR1, FR2, FR3, and FR4, respectively, of a nanobody comprising the amino acid sequence of SEQ ID NO:53. The FR1, FR2, FR3, and FR4 may comprise the amino acid sequence of SEQ ID NOS:221-224, respectively or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:221-224, respectively. The FR1, FR2, FR3, and FR4 may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the amino acid sequence of SEQ ID NOS:221-224, respectively.

[0017] HCDR1, HCDR2, and HCDR3 may have or have at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:2. LCDR1, LCDR2, and LCDR3 may have or have at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:3. The VH may comprise a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:2 and the VL may comprise a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:3. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:4-9, respectively. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NOS:4-9, respectively. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise the amino acid sequence of SEQ ID NOS:4-9, respectively.202666783.1 - 10 -

[0018] The VH may comprise an amino acid sequence with at least 70% sequence identity to SEQ ID NO:2 and / or the VL may comprise an amino acid sequence with at least 70% sequence identity to SEQ ID NO:3. The VH may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:2 and / or the VL may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:3. The VH may comprise the amino acid sequence of SEQ ID NO:2 and / or the VL may comprise the amino acid sequence of SEQ ID NO:3.

[0019] The antibody or antigen binding fragment may comprise a heavy chain framework region (HFR) 1, HFR2, HFR3, and HFR4 and light chain framework region (LFR) 1, LFR2, LFR3, and LFR4, and wherein the HFR1, HFR2, HFR3, and HFR4 comprises an amino acid sequence with at least 80% sequence identity to an HFR1, HFR2, HFR3, and HFR4, respectively, of a VH comprising the amino acid sequence of SEQ ID NO:2, and the LFR1, LFR2, LFR3, and LFR4 comprises an amino acid sequence with at least 80% sequence identity to the LFR1, LFR2, LFR3, and LFR4, respectively, of a VL comprising the amino acid sequence of SEQ ID NO:3. The HFR1, HFR2, HFR3, and HFR4 may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to an HFR1, HFR2, HFR3, and HFR4, respectively, of a VH comprising the amino acid sequence of SEQ ID NO:2, and the LFR1, LFR2, LFR3, and LFR4 may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the LFR1, LFR2, LFR3, and LFR4, respectively, of a VL comprising the amino acid sequence of SEQ ID NO:3. The HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4 may comprise the amino acid sequence of SEQ ID NOS:10-17, respectively, or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:10-17, respectively. The HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4 may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93,202666783.1 - 11 -94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the amino acid sequence of SEQ ID NOS:10-17, respectively.

[0020] The HCDR1, HCDR2, and HCDR3 may have or have at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:19. The LCDR1, LCDR2, and LCDR3 may have or have at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:20. The VH may comprise a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:19 and the VL may comprise a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:20. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:21-26, respectively. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NOS:21-26, respectively. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise the amino acid sequence of SEQ ID NOS:21-26, respectively.

[0021] The VH may comprise an amino acid sequence with at least 80% sequence identity to SEQ ID NO:19 and / or the VL may comprise an amino acid sequence with at least 80% sequence identity to SEQ ID NO:20. The VH may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:19 and / or the VL may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:20. The VH may comprise the amino acid sequence of SEQ ID NO:19 and / or the VL may comprise the amino acid sequence of SEQ ID NO:20.202666783.1 - 12 -

[0022] The antibody or antigen binding fragment may comprise a heavy chain framework region (HFR) 1, HFR2, HFR3, and HFR4 and light chain framework region (LFR) 1, LFR2, LFR3, and LFR4, and wherein the HFR1, HFR2, HFR3, and HFR4 comprises an amino acid sequence with at least 80% sequence identity to an HFR1, HFR2, HFR3, and HFR4, respectively, of a VH comprising the amino acid sequence of SEQ ID NO:19, and the LFR1, LFR2, LFR3, and LFR4 comprises an amino acid sequence with at least 80% sequence identity to the LFR1, LFR2, LFR3, and LFR4, respectively, of a VL comprising the amino acid sequence of SEQ ID NO:20. The HFR1, HFR2, HFR3, and HFR4 may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to an HFR1, HFR2, HFR3, and HFR4, respectively, of a VH comprising the amino acid sequence of SEQ ID NO:19, and the LFR1, LFR2, LFR3, and LFR4 may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the LFR1, LFR2, LFR3, and LFR4, respectively, of a VL comprising the amino acid sequence of SEQ ID NO:20.

[0023] The HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4 may comprise the amino acid sequence of SEQ ID NOS:27-34, respectively or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:27-34, respectively. The HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4 may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the amino acid sequence of SEQ ID NOS:27-34, respectively.

[0024] The antibody may comprise a VH having at least 70% sequence identity to SEQ ID NO:19 and / or a VL having at least 70% sequence identity to SEQ ID NO:20. The antibody may comprise a VH that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the amino acid sequence of SEQ ID NO:19 and / or a VL that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the amino acid202666783.1 - 13 -sequence of SEQ ID NO:20. The antibody may comprise a VH of SEQ ID NO:19 and / or a VL of SEQ ID NO:20.

[0025] The antibody may be further defined as human, chimeric, or humanized. The antibody or antigen binding fragment may be further defined as antagonistic. The antibody may be one that includes or excludes a human antibody, humanized antibody, recombinant antibody, chimeric antibody, an antibody derivative, a veneered antibody, a diabody, a monoclonal antibody, a single domain antibody, or a single chain antibody. The antigen binding fragment may include or exclude a single chain variable fragment (scFv), F(ab’)2, Fab’, Fab, Fv, or rIgG. The antigen binding fragment may comprise a scFv. The antigen binding fragment may comprise the amino acid sequence of SEQ ID NO:1 or 18 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:1 or 18. The antigen binding fragment may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:1 or 18. The polypeptide may comprise or exclude at least two antigen binding fragments, wherein each antigen binding fragment is independently selected from an antigen binding fragment of the disclosure. The polypeptide may be defined multivalent. Multivalent polypeptides may be excluded. The polypeptide may be a bispecific polypeptide.

[0026] The VH may be amino proximal to the VL. The VL may be amino proximal to the VH. A first region is carboxy-proximal to a second region when the first region is attached to the carboxy terminus of the second region. There may be further intervening amino acid residues between the first and second regions. Thus, the regions need not be immediately adjacent, unless specifically specified as not having intervening amino acid residues. The term “amino-proximal” is similarly defined in that a first region is amino-proximal to a second region when the first region is attached to the amino terminus of the second region. Similarly, there may be further intervening amino acid residues between the first and second regions unless stated otherwise. The antigen binding fragment may comprise a VH and a VL and a linker between the VH and VL. The linker may comprise or exclude a glycine serine linker. The linker may comprise or exclude (G4S)n, wherein n is an integer selected from 1-10. The linker may comprise, exclude, or consist of SEQ ID NO:119 or 121. The linker may comprise or exclude SEQ ID NO:120.

[0027] The antigen binding domain may comprise the amino acid sequence of SEQ ID NO:35 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:35. The antigen binding domain may comprise an amino acid sequence that has or has at least 60,202666783.1 - 14 -61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:35. The antigen binding domain may comprise the amino acid sequence of SEQ ID NO:44 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:44. The antigen binding domain may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:44.

[0028] The polypeptide may comprise or exclude a signal peptide. The signal peptide may comprise or exclude SEQ ID NO:122 or 123 or an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:122 or 123.

[0029] The transmembrane domain may include or exclude an α or β chain of the T cell receptor or a transmembrane domain from CD28, CD3-epsilon, CD45, CD4, CD5, CD8-α, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD123, CD134, CD137 or CD154. The transmembrane domain may be a CD28 transmembrane domain. The transmembrane domain may comprise SEQ ID NO:130 or 131 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:130 or 131. The transmembrane domain may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:130 or 131.

[0030] The intracellular signaling domain may comprise or exclude a primary signaling domain. The primary signaling domain may be a CD3ζ (zeta) signaling domain. The primary signaling domain may comprise SEQ ID NO:134 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:134. The primary signaling domain may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:134. The intracellular signaling domain may comprise or exclude a costimulatory domain. The costimulatory domain may comprise or exclude a signaling domain from 4-1BB (CD137), CD28, IL-15Rα, OX40, CD2, CD27, CDS, ICAM-1, LFA-1 (CD11a / CD18), or ICOS (CD278). The costimulatory domain may comprise a CD28 or 4-1BB signaling domain. The costimulatory domain may comprise SEQ ID NO:132 or 133 or an amino acid sequence having202666783.1 - 15 -at least 80% sequence identity to SEQ ID NO:132 or 133. The costimulatory domain may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:132 or 133.

[0031] The antigen binding domain and the transmembrane domain may be separated by a hinge region. The hinge region may comprise or exclude an IgG4 hinge, a CD8α hinge, an IgG1 hinge, or a CD34 hinge. The hinge region may be between 8 and 300 amino acids in length. The hinge region may be, be at least, or be at most 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, or 300 amino acids in length or any derivable range therein. The hinge region may comprise one of SEQ ID NOS:126-129 or an amino acid sequence having at least 80% sequence identity to one of SEQ ID NOS:126-129. The hinge region may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to one of SEQ ID NOS:126-129.

[0032] The nucleic acid may be DNA. The nucleic acid may be RNA. The nucleic acid may be an expression construct. The expression construct may include or exclude a plasmid or viral vector. The viral vector may include or exclude a vector derived from a retrovirus or a vector derived from a lentivirus. Cells of the disclosure include or exclude those in which the viral vector has integrated into the cell’s genome. The nucleic acid may include or exclude one202666783.1 - 16 -or more promoter(s) that direct the expression of the suicide gene product, CAR, IL-12 polypeptide, IFN-β polypeptide, CXCL9 polypeptide, and / or IL-18 polypeptide. The promoter may be constitutive. The promoter may be conditional. The promoter may be one that responds positively to at least one cytokine or to T-cell activation. The at least one cytokine may include or exclude TGF- β, IL-6, TNF α, IFN-γ, IL-1β, IL-2, IL-8, or IL-10 or the promoter responds positively to NFAT or NF-κB. The nucleic acid may encode for a chimeric antigen receptor (CAR), wherein the CAR comprises: (a) an antigen binding domain (b) a transmembrane domain; and (c) an intracellular signaling domain.

[0033] The cell(s) may comprise or exclude a T cell, a natural killer (NK) cell, a natural killer T cell (NKT), an invariant natural killer T cell (iNKT), stem cell, lymphoid progenitor cell, peripheral blood mononuclear cell (PBMC), bone marrow cell, fetal liver cell, embryonic stem cell, cord blood cell, induced pluripotent stem cell (iPS cell). The cell may comprise or exclude a T cell or an NK cell. The T cell may comprise a naïve memory T cell. The naïve memory T cell may comprise a CD4+ or CD8+ T cell. The T cell may comprise or exclude a T cell from a population of CD14 depleted, CD25 depleted, and / or CD62L enriched PBMCs. The cell may be further defined as a human cell.

[0034] Cells include those that have been infected with a virus comprising a nucleic acid of the disclosure. The virus may comprise lentivirus or a lentiviral-derived virus or vector. The cell may be one that is not yet a T cell or NK cell, the method further comprising culturing the cell under conditions that promote the differentiation of the cell into a T cell or an NK cell. The methods may comprise culturing the cell under conditions to expand the cell before and or after introducing the nucleic acid into the cell. The cell may be cultured with serum-free medium. The cell or cells of the disclosure may comprises a nucleic acid encoding a CAR or TCR.

[0035] The nucleic acid may further encode for a suicide gene product. The cell may further comprise a nucleic acid encoding for a suicide gene product. The suicide gene product may include or exclude herpes simplex virus thymidine kinase (HSV-TK), purine nucleoside phosphorylase (PNP), cytosine deaminase (CD), carboxypetidase G2, cytochrome P450, linamarase, β-lactamase, nitroreductase (NTR), carboxypeptidase A, split Cas9, truncated EGFR, or inducible caspase 9. The nucleic acid may further encode for a suicide gene product. Suicide gene products provide for a safety switch that can be activated to eliminate the cell from the body in case of adverse reactions. A suicide gene may be enzyme-based, meaning the gene product of the suicide gene is an enzyme and the suicide function depends on enzymatic activity. One or more suicide genes may be utilized in a single cell or clonal population. The202666783.1 - 17 -suicide gene may encode herpes simplex virus thymidine kinase (HSV-TK), truncated EGFR, purine nucleoside phosphorylase (PNP), cytosine deaminase (CD), carboxypetidase G2, cytochrome P450, linamarase, beta-lactamase, nitroreductase (NTR), carboxypeptidase A, or inducible caspase 9. Methods in the art for suicide gene usage may be employed, such as in U.S. Patent No. 8628767, U.S. Patent Application Publication 20140369979, U.S. 20140242033, and U.S. 20040014191, all of which are incorporated by reference in their entirety. Caspase suicide genes are described in Di Stasi A, et al., N Engl J Med. 2011 Nov 3;365(18):1673-83, which is incorporated by reference. Using truncated EGFR as a safety switch is further described in Kao RL, et al., Hum Gene Ther.2019 Apr;30(4):413-428, which is incorporated by reference. The TK gene may be a viral TK gene, .i.e., a TK gene from a virus. The TK gene may be a herpes simplex virus TK gene. The suicide gene product may be activated by a substrate. Thymidine kinase is a suicide gene product that is activated by ganciclovir, penciclovir, or a derivative thereof. The suicide gene product may be encoded by the same or a different nucleic acid molecule encoding for the CAR and / or other components listed throughout. In alternative embodiments, the cell does not express an exogenous suicide gene.

[0036] The nucleic acid may encode for an IL-12 polypeptide and an IL-18 polypeptide. The nucleic acid may encode for an IL-12, IFN-β, and / or CXCL9 polypeptide. The cell may comprise a heterologous nucleic acid that encodes for an IL-12 polypeptide and a heterologous nucleic acid that encodes for an IL-18 polypeptide. The cell may comprise a heterologous nucleic acid encoding an IL-12 polypeptide, a heterologous nucleic acid encoding an IFN-β polypeptide and a heterologous nucleic acid encoding a CXCL9 polypeptide. The IL-12 polypeptide may comprise a p40 and / or p35 subunit. The IL-12 polypeptide may comprise a single-chain IL-12 polypeptide. The p40 subunit may be amino-proximal to the p35 subunit. The p35 subunit may be amino-proximal to the p40 subunit. The IL-12 polypeptide is defined as a polypeptide from IL-12 or variant that is functional. The IL-12 polypeptide may be defined as a polypeptide from IL-12 or variant that stimulates the production of IFN-γ and / or TNF-α from T cells and natural killer (NK) cells. The IL-12 polypeptide may be defined as a polypeptide from IL-12 or variant that reduces IL-4 mediated suppression of IFN-γ. The IL-18 polypeptide is defined as a polypeptide from IL-18 or variant that is functional. The IL-18 polypeptide may be defined as a polypeptide from IL-18 or variant that activates proinflammatory molecules and / or the NF-κB pathway. The IL-18 polypeptide may be defined as a polypeptide from IL-18 or variant that induces cell-mediated immunity and / or induces IFNγ production. The IL-18 polypeptide may be defined as a polypeptide from IL-18 or variant202666783.1 - 18 -that inhibits IL-4 dependent IgE and IgG1 production and / or enhances IgG2a production in B cells. These functions of IL-18 may be performed in combination with the IL-12 polypeptide. The IFN-β polypeptide is defined as a polypeptide from IFN-β or variant that is functional. The IFN-β polypeptide may be defined as one that stimulates innate immunity. The CXCL9 polypeptide is defined as a polypeptide from CXCL9 or variant that is functional. The CXCL9 polypeptide may be defined as one that interacts with CXCR3. The CXCL9 polypeptide may be defined as one that stimulates immune cells and / or stimulates the production of IFN-g, TNF- a, and / or IL-2 from immune cells.

[0037] The nucleic acid may encode for a linker between the p40 and p35 subunit of the IL-12 polypeptide. The nucleic acid may comprise one or more promoter(s) that direct the expression of the suicide gene product, CAR, polypeptide, IL-12 polypeptide, IL-18 polypeptide, IFN-β polypeptide, CXCL9 polypeptide, and / or anti-VEGF antigen binding fragment. The IL-12 polypeptide, IFN-β polypeptide, CXCL9 polypeptide, and / or IL-18 polypeptide may be derived from a human IL-12 polypeptide, human IFN-β polypeptide, human CXCL9 polypeptide, and / or human IL-18 polypeptide The promoter may be constitutive. The promoter may be conditional. The promoter may be one that responds positively to at least one cytokine or to T-cell activation. The at least one cytokine may include or exclude TGF- β, IL-6, TNF α, IFN-γ, IL-1β, IL-2, IL-8, or IL-10 or the promoter responds positively to NFAT or NF-κB.

[0038] The IL-18 polypeptide may comprise a decoy-resistant IL-18 polypeptide. The p40 polypeptide may comprise the amino acid sequence of SEQ ID NO:236 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:236. The p40 polypeptide may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:236. The p35 polypeptide may comprise the amino acid sequence of SEQ ID NO:237 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:237. The p35 polypeptide may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:237. The IL-12 polypeptide may comprise the amino acid sequence of SEQ ID NO:230 or 234 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:230 or 234. The IL-12 polypeptide may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82,202666783.1 - 19 -83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:230 or 234. The IL-18 polypeptide may comprise the amino acid sequence of SEQ ID NO:229, 232, or 238-240 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:229, 232, or 238-240. The IL-18 polypeptide may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:229, 232, or 238-240.

[0039] The IFN-β polypeptide may comprise the amino acid sequence of SEQ ID NO:253 or 255 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:253 or 255. The IFN-β polypeptide may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:253 or 255. The CXCL9 polypeptide may comprise the amino acid sequence of SEQ ID NO:257 or 259 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:257 or 259. The CXCL9 polypeptide may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:257 or 259.

[0040] The nucleic acid may encode for a signal sequence at the amino-terminus of the IL- 12, IFN-β, CXCL9 and / or IL-18 polypeptide. The signal sequence may comprise SEQ ID NO:122, 123, 235, 241, 254, 256, 258, and / or 260. The IL-12 polypeptide, the IL-18 polypeptide, and / or anti-VEGF antigen binding fragment may be expressed from one nucleic acid. The nucleic acid may be one that expresses a polypeptide comprising the IL-12 polypeptide, the IL-18 polypeptide, and / or anti-VEGF antigen binding fragment. The IL-12 polypeptide, IFN-β polypeptide, CXCL9 polypeptide, and / or anti-VEGF antigen binding fragment may be expressed from one nucleic acid. The nucleic acid may be one that expresses a polypeptide comprising the IL-12, IFN-β, CXCL9 polypeptide, and / or anti-VEGF antigen binding fragment. The nucleic acid may encode for a self-cleaving peptide or internal ribosome entry sequence (IRES), wherein the self-cleaving peptide or IRES is between the IL-12, IL-18, CXCL9, and / or IFN-β polypeptides. The self-cleaving peptide may comprise SEQ ID NO:135 or 136.

[0041] The anti-VEGF antigen binding fragment, such as the anti-VEGF scFv, when expressed and secreted from a cell, such as a T cell, may be used in adoptive cell therapy, and202666783.1 - 20 -may be administered in combination with a cytokine (either expressed by the same cell, expressed by another cell, or administered systemically). This may provide the unexpected effect of reducing the toxicity of the cytokine.

[0042] In some aspects, the anti-VEGF antibodies or antigen binding fragments of the disclosure may be used in a method comprising complement-dependent cytotoxicity or antibody-dependent cellular cytotoxicity.

[0043] The antigen binding domain may comprise or exclude an anti-TYRP-1, anti-IL- 13Rα2, anti-TGF-β, anti-CLDN6, anti-GPC3, or anti-MSLN antigen binding domain. The antigen binding domain may comprise an anti-IL-13Rα2 comprising an IL-13 mutein polypeptide. The IL-13 mutein polypeptide may comprise the amino acid sequence of SEQ ID NO:115 or 233, or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:115 or 233. The IL-13 mutein polypeptide may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:115 or 233.

[0044] The antigen binding domain may comprise anti-TYRP-1. The antigen binding domain may comprise a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH may comprise a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:62 and wherein the light chain variable region may comprise a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:63. The VH may comprise a HCDR1, HCDR2, and HCDR3 that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:62. The light chain variable region may comprise a LCDR1, LCDR2, and LCDR3 that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:63. The VH may comprise a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:62 and wherein the VL may comprise a LCDR1, LCDR2,202666783.1 - 21 -and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:63. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:64-69. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NOS:64-69. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise the amino acid sequence of SEQ ID NOS:64-69, respectively. The VH may comprise an amino acid sequence with at least 70% sequence identity to SEQ ID NO:62 and / or the VL may comprise an amino acid sequence with at least 70% sequence identity to SEQ ID NO:63. The VH may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:62. The VL may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:63. The VH may comprise the amino acid sequence of SEQ ID NO:62. The VL may comprise the amino acid sequence of SEQ ID NO:63.

[0045] The antigen binding domain may comprise anti-TGF-β. The antigen binding domain may comprise a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH may comprise a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:71 and wherein the light chain variable region may comprise a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:72. The VH may comprise a HCDR1, HCDR2, and HCDR3 that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:71. The light chain variable region may comprise a LCDR1, LCDR2, and LCDR3 that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85,202666783.1 - 22 -86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:72. The VH may comprise a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:71. The VL may comprise a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:72. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:73-78, respectively. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NOS:73-78, respectively. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise the amino acid sequence of SEQ ID NOS:73-78, respectively. The VH may comprise an amino acid sequence with at least 70% sequence identity to SEQ ID NO:71. The VL may comprise an amino acid sequence with at least 70% sequence identity to SEQ ID NO:72. The VH may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:71. The VL may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:72. The VH may comprise the amino acid sequence of SEQ ID NO:71. The VL may comprise the amino acid sequence of SEQ ID NO:72.

[0046] The antigen binding domain may comprise a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH may comprise a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:80 and wherein the light chain variable region may comprise a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:81. The VH may comprise a HCDR1, HCDR2, and HCDR3 that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86,202666783.1 - 23 -87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:80. The light chain variable region may comprise a LCDR1, LCDR2, and LCDR3 that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:81. The VH may comprise a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:80. The VL may comprise a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:81. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:82-87, respectively. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NOS:82-87, respectively. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise the amino acid sequence of SEQ ID NOS:82-87, respectively. The VH may comprise an amino acid sequence with at least 70% sequence identity to SEQ ID NO:80. The VL may comprise an amino acid sequence with at least 70% sequence identity to SEQ ID NO:81. The VH may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:80. The VL may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:81. The VH may comprise the amino acid sequence of SEQ ID NO:80. The VL may comprise the amino acid sequence of SEQ ID NO:81.

[0047] The antigen binding domain may comprise a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH may comprise a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:89 and wherein the light chain variable region may comprise a light chain complementarity202666783.1 - 24 -determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:90. The VH may comprise a HCDR1, HCDR2, and HCDR3 that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:89. The light chain variable region may comprise a LCDR1, LCDR2, and LCDR3 that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:90. The VH may comprise a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:89. The VL may comprise a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:90. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:91-96, respectively. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NOS:91-96, respectively. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise the amino acid sequence of SEQ ID NOS:91-96, respectively. The VH may comprise an amino acid sequence with at least 70% sequence identity to SEQ ID NO:89. The VL may comprise an amino acid sequence with at least 70% sequence identity to SEQ ID NO:90. The VH may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:89. The VL may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:90. The VH may comprise the amino acid sequence of SEQ ID NO:89. The VL may comprise the amino acid sequence of SEQ ID NO:90.202666783.1 - 25 -

[0048] The antigen binding domain may comprise anti-CLDN6. The antigen binding domain may comprise a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH may comprise a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:98 and wherein the light chain variable region may comprise a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:99. The VH may comprise a HCDR1, HCDR2, and HCDR3 that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:98. The VL may comprise a LCDR1, LCDR2, and LCDR3 that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:99. The VH may comprise a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:98. The VL may comprise a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:99. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:100-103, the sequence TSN, and SEQ ID NO:105, respectively. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NOS:100-103, the sequence TSN, and SEQ ID NO:105, respectively. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise the amino acid sequence of SEQ ID NOS:100-103, the sequence TSN, and SEQ ID NO:105, respectively. The VH may comprise an amino acid sequence with at least 70% sequence identity to SEQ ID NO:98. The VL may comprise an amino acid sequence with at least 70% sequence identity to SEQ ID NO:99. The VH may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence202666783.1 - 26 -identity (or any derivable range therein) to SEQ ID NO:98. The VL may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:99. The VH may comprise the amino acid sequence of SEQ ID NO:98. The VL may comprise the amino acid sequence of SEQ ID NO:99.

[0049] The antigen binding domain may comprise anti-GPC3. The antigen binding domain may comprise a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH may comprise a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:107 and wherein the VL may comprise a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:108. The VH may comprise a HCDR1, HCDR2, and HCDR3 that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:107. The VL may comprise a LCDR1, LCDR2, and LCDR3 that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:108. The VH may comprise a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:107. The VL may comprise a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:108. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:109-114, respectively. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NOS:109-114, respectively. The HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 may each comprise the amino acid sequence of SEQ ID NOS:109-114, respectively. The VH202666783.1 - 27 -may comprise an amino acid sequence with at least 70% sequence identity to SEQ ID NO:107. The VL may comprise an amino acid sequence with at least 70% sequence identity to SEQ ID NO:108. The VH may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:107. The VL may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to SEQ ID NO:108. The VH may comprise the amino acid sequence of SEQ ID NO:107. The VL may comprise the amino acid sequence of SEQ ID NO:108.

[0050] The antigen binding domain may comprise the amino acid sequence of one of SEQ ID NOS:35, 44, 53, 57, 61, 70, 79, 88, 97, or 106 or an amino acid sequence having at least 80% sequence identity to one of SEQ ID NOS:35, 44, 53, 57, 61, 70, 79, 88, 97, or 106. The antigen binding domain may comprise an amino acid sequence that has or has at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity (or any derivable range therein) to one of SEQ ID NOS:35, 44, 53, 57, 61, 70, 79, 88, 97, or 106.

[0051] Any of the cell populations may comprise at least, about, or at most, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 1 x 103, 2 x 103, 3 x 103, 4 x 103, 5 x 103, 6 x 103, 7 x 103, 8 x 103, 9 x 103, 1 x 104, 2 x 104, 3 x 104, 4 x 103,5 x 104,6 x 104, 7 x 104, 8 x 104, 9 x 104, 1 x 105, 2 x 105, 3 x 105, 4 x 105,5 x 105, 6 x 105, 7 x 105, 8 x 105, 9 x 105, 1 x 106, 2 x 106, 3 × 106, 4 × 106, 5 × 106, 6 × 106, 7 × 106, 8 × 106, 9 × 106, 1 × 107, 2 × 107, 3 × 107, 4 × 107, 5 × 107, 6 × 107, 7 × 107, 8 × 107, 9 × 107, 1 × 108, 2 × 108, 3 × 108, 4 × 108, 5 × 108, 6 × 108, 7 × 108, 8 × 108, 9 × 108, 1 × 109, 2 × 109, 3 × 109, 4 × 109, 5 × 109, 6 × 109, 7 × 109, 8 × 109, 9 × 109, 1 × 1010, 2 × 1010, 3 × 1010, 4 × 1010, 5 × 1010, 6 × 1010, 7 × 1010, 8 × 1010, 9 × 1010, 1 × 1011, 2 × 1011, 3 × 1011, 4 × 1011, 5 × 1011, 6 × 1011, 7 × 1011, 8 × 1011, 9 × 1011, 1 × 1012, 2 × 1012, 3 × 1012, 4 × 1012, 5 × 1012, 6 × 1012, 7 × 1012, 8 × 1012, 9 × 1012, 1 × 1013, 2 × 1013, 3 × 1013, 4 × 1013, 5 × 1013, 6 × 1013, 7 × 1013, 8 × 1013, 9 × 1013, or 1 × 1014cells, or any derivable range therein. The cells may be a population of cells comprising 103-1012cells. The cells may be a population of cells comprising 103-108cells.

[0052] The patient may be one that has and / or has been determined to have a MSLN+ cancer. The cancer may comprise or exclude ovarian cancer, stomach cancer, colorectal cancer, endometrial cancer, uterine cancer, acute myeloid leukemia, pancreatic cancer, lung cancer, or202666783.1 - 28 -mesothelioma. The subject or patient may be one that has ovarian, melanoma, glioma, glioblastoma, sarcoma, hepatocellular carcinoma, lung cancer, non-small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, breast cancer, neuroblastoma, thyroid cancer, gastric cancer, ovarian cancer, testis cancer, or uterine cancer. The methods may further comprise administering an additional therapy to the patient. The additional therapy may comprise or exclude an immunotherapy, anti-angiogenic therapy, chemotherapy, surgery, radiotherapy, neoantigen therapy, or vaccination. The additional therapy may comprise or exclude an immunotherapy and wherein the immunotherapy comprises immune checkpoint inhibitor therapy or bispecific T cell engagers. The cells may be autologous or non-autologous. The subject or patient may be a human subject. The subject or patient may be further defined as a mammal, a non-human primate, a pig, horse, mouse, rat, rabbit, or dog. The term “subject” and “patient” is used interchangeably herein.

[0053] When reciting that the CDR1, CDR2, and CDR3 has at least 80% sequence identity to the CDR1, CDR2, and CDR3 or a specific amino acid sequence, one skilled in the art would understand that each of CDR1, CDR2, and CDR3 can share at least 80% identity to CDR, CDR2, and CDR3, respectively of the specific recited amino acid sequences.

[0054] The therapeutic agents of the disclosure may be used for in vivo, in vitro, or ex vivo administration. The route of administration of the therapies may be, for example, intracutaneous, subcutaneous, intravenous, local, topical, and intraperitoneal administrations.

[0055] In some embodiments, the disclosed methods are directed to methods for treating cancer. The cancer may be a solid tumor, metastatic cancer, or non-metastatic cancer. In certain embodiments, the cancer may be recurrent, metastatic, relapsed, or of a Stage I, II, III, or IV.

[0056] Also included in the disclosure are the embodiments and combinations of embodiments described below and herein.

[0057] Embodiment one is a chimeric polypeptide comprising: (a) an antigen binding domain comprising a nanobody, wherein the nanobody comprises a complementarity determining region (CDR) 1, CDR2, and CDR3 having at least 80% sequence identity to the CDR1, CDR2, CDR3 from the nanobody of SEQ ID NO:57; (b) a transmembrane domain; and (c) an intracellular signaling domain.

[0058] Embodiment two is the chimeric polypeptide of embodiment 1, wherein the nanobody comprises a CDR1, CDR2, and CDR3 having the amino acid sequence of the CDR1, CDR2, and CDR3, respectively, from the nanobody of SEQ ID NO:57.202666783.1 - 29 -

[0059] Embodiment three is the chimeric polypeptide of embodiment 1 or embodiment 2, wherein the CDR1, CDR2, and CDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:58-60, respectively.

[0060] Embodiment four is the chimeric polypeptide of any one of embodiments 1 to 3, wherein the CDR1, CDR2, and CDR3 each comprise the amino acid sequence of SEQ ID NOS:58-60, respectively.

[0061] Embodiment five is the chimeric polypeptide of any one of embodiments 1 to 4, wherein the nanobody comprises a framework region (FR) 1, FR2, FR3, and FR4, wherein the FR1, FR2, FR3, and FR4 comprises an amino acid sequence with at least 80% sequence identity to an FR1, FR2, FR3, and FR4, respectively, of a nanobody comprising the amino acid sequence of SEQ ID NO:57.

[0062] Embodiment six is the chimeric polypeptide of embodiment 5, wherein the FR1, FR2, FR3, and FR4 comprises the amino acid sequence of SEQ ID NOS:225-228, respectively or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:225-228, respectively.

[0063] Embodiment seven is the chimeric polypeptide of any one of embodiments 1 to 6, wherein the antigen binding domain and / or nanobody comprises the amino acid sequence of SEQ ID NO:57 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:57.

[0064] Embodiment eight is a chimeric polypeptide comprising: (a) an antigen binding domain comprising a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:36 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:37; (b) a transmembrane domain; and (c) an intracellular signaling domain.

[0065] Embodiment nine is the chimeric polypeptide of embodiment 8, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:36 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:37.202666783.1 - 30 -

[0066] Embodiment 10 is the chimeric polypeptide of embodiment 8 or embodiment 9, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:38-43, respectively.

[0067] Embodiment 11 is the chimeric polypeptide of any one of embodiments 8 to 10, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:38-43, respectively.

[0068] Embodiment 12 is the chimeric polypeptide of any one of embodiments 8 to 11, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:36 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:37.

[0069] Embodiment 13 is the chimeric polypeptide of embodiment 12, wherein the VH comprises the amino acid sequence of SEQ ID NO:36 and / or the VL comprises the amino acid sequence of SEQ ID NO:37.

[0070] Embodiment 14 is the chimeric polypeptide of any one of embodiments 8 to 13, wherein the VH comprises a heavy chain framework region (HFR) 1, HFR2, HFR3, and HFR4 and the VL comprises a light chain framework region (LFR) 1, LFR2, LFR3, and LFR4, and wherein the HFR1, HFR2, HFR3, and HFR4 comprises an amino acid sequence with at least 80% sequence identity to an HFR1, HFR2, HFR3, and HFR4, respectively, of a VH comprising the amino acid sequence of SEQ ID NO:36, and the LFR1, LFR2, LFR3, and LFR4 comprises an amino acid sequence with at least 80% sequence identity to the LFR1, LFR2, LFR3, and LFR4, respectively, of a VL comprising the amino acid sequence of SEQ ID NO:37.

[0071] Embodiment 15 is the chimeric polypeptide of embodiment 14, wherein the HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4 comprises the amino acid sequence of SEQ ID NOS:208-215, respectively or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:208-215, respectively.

[0072] Embodiment 16 is the chimeric polypeptide of any one of embodiments 8 to 15, wherein the VH is amino proximal to the VL.

[0073] Embodiment 17 is the chimeric polypeptide of any one of embodiments 8 to 15, wherein the VL is amino proximal to the VH.

[0074] Embodiment 18 is the chimeric polypeptide of any one of embodiments 8 to 17, wherein the antigen binding domain comprises a linker between the VH and VL.

[0075] Embodiment 19 is the chimeric polypeptide of embodiment 18, wherein the linker comprises a glycine serine linker.202666783.1 - 31 -

[0076] Embodiment 20 is the chimeric polypeptide of embodiment 19, wherein the linker comprises (G4S)n, wherein n is an integer selected from 1-10.

[0077] Embodiment 21 is the chimeric polypeptide of embodiment 20, wherein the linker comprises or consists of SEQ ID NO:119 or 121.

[0078] Embodiment 22 is the chimeric polypeptide of embodiment 18, wherein the linker comprises SEQ ID NO:120.

[0079] Embodiment 23 is the chimeric polypeptide of any one of embodiments 8 to 22, wherein the antigen binding domain comprises the amino acid sequence of SEQ ID NO:35 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:35.

[0080] Embodiment 24 is a chimeric polypeptide comprising: (a) an antigen binding domain comprising a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:45 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:46;(b) a transmembrane domain; and (c) an intracellular signaling domain.

[0081] Embodiment 25 is the chimeric polypeptide of embodiment 24, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:45 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:46.

[0082] Embodiment 26 is the chimeric polypeptide of embodiment 24 or embodiment 25, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:47-52, respectively.

[0083] Embodiment 27 is the chimeric polypeptide of any one of embodiments 24 to 26, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:47-52, respectively.

[0084] Embodiment 28 is the chimeric polypeptide of any one of embodiments 24 to 27, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ202666783.1 - 32 -ID NO:45 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:46.

[0085] Embodiment 29 is the chimeric polypeptide of embodiment 28, wherein the VH comprises the amino acid sequence of SEQ ID NO:45 and / or the VL comprises the amino acid sequence of SEQ ID NO:46.

[0086] Embodiment 30 is the chimeric polypeptide of any one of embodiments 24 to 29, wherein the VH comprises a heavy chain framework region (HFR) 1, HFR2, HFR3, and HFR4 and the VL comprises a light chain framework region (LFR) 1, LFR2, LFR3, and LFR4, and wherein the HFR1, HFR2, HFR3, and HFR4 comprises an amino acid sequence with at least 80% sequence identity to an HFR1, HFR2, HFR3, and HFR4, respectively, of a VH comprising the amino acid sequence of SEQ ID NO:45, and the LFR1, LFR2, LFR3, and LFR4 comprises an amino acid sequence with at least 80% sequence identity to the LFR1, LFR2, LFR3, and LFR4, respectively, of a VL comprising the amino acid sequence of SEQ ID NO:46.

[0087] Embodiment 31 is the chimeric polypeptide of embodiment 30, wherein the HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4 comprises the amino acid sequence of SEQ ID NOS:216, 209, 217, 211, 218, 219, 220, and 215, respectively or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:216, 209, 217, 211, 218, 219, 220, and 215, respectively.

[0088] Embodiment 32 is the chimeric polypeptide of any one of embodiments 24 to 31, wherein the VH is amino proximal to the VL.

[0089] Embodiment 33 is the chimeric polypeptide of any one of embodiments 24 to 31, wherein the VL is amino proximal to the VH.

[0090] Embodiment 34 is the chimeric polypeptide of any one of embodiment 24 to 33, wherein the antigen binding domain comprises a linker between the VH and VL.

[0091] Embodiment 35 is the chimeric polypeptide of embodiment 34, wherein the linker comprises a glycine serine linker.

[0092] Embodiment 36 is the chimeric polypeptide of embodiment 35, wherein the linker comprises (G4S)n, wherein n is an integer selected from 1-10.

[0093] Embodiment 37 is the chimeric polypeptide of embodiment 36, wherein the linker comprises or consists of SEQ ID NO:119 or 121.

[0094] Embodiment 38 is the chimeric polypeptide of embodiment 34, wherein the linker comprises SEQ ID NO:120.202666783.1 - 33 -

[0095] Embodiment 39 is the chimeric polypeptide of any one of embodiments 24 to 38, wherein the antigen binding domain comprises the amino acid sequence of SEQ ID NO:44 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:44.

[0096] Embodiment 40 is a chimeric polypeptide comprising: (a) an antigen binding domain comprising a nanobody, wherein the nanobody comprises a complementarity determining region (CDR) 1, CDR2, and CDR3 having at least 80% sequence identity to the CDR1, CDR2, CDR3 from the nanobody of SEQ ID NO:53; (b) a transmembrane domain; and (c) an intracellular signaling domain.

[0097] Embodiment 41 is the chimeric polypeptide of embodiment 40, wherein the nanobody comprises a CDR1, CDR2, and CDR3 having the amino acid sequence of the CDR1, CDR2, and CDR3, respectively, from the nanobody of SEQ ID NO:53.

[0098] Embodiment 42 is the chimeric polypeptide of embodiment 40 or embodiment 41, wherein the CDR1, CDR2, and CDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:54-56, respectively.

[0099] Embodiment 43 is the chimeric polypeptide of any one of embodiments 40 to 42, wherein the CDR1, CDR2, and CDR3 each comprise the amino acid sequence of SEQ ID NOS:54-56, respectively.

[0100] Embodiment 44 is the chimeric polypeptide of any one of embodiments 40 to 43, wherein the nanobody comprises a framework region (FR) 1, FR2, FR3, and FR4, wherein the FR1, FR2, FR3, and FR4 comprises an amino acid sequence with at least 80% sequence identity to an FR1, FR2, FR3, and FR4, respectively, of a nanobody comprising the amino acid sequence of SEQ ID NO:53.

[0101] Embodiment 45 is the chimeric polypeptide of embodiment 44, wherein the FR1, FR2, FR3, and FR4 comprises the amino acid sequence of SEQ ID NOS:221-224, respectively or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:221-224, respectively.

[0102] Embodiment 46 is the chimeric polypeptide of any one of embodiments 40 to 45, wherein the antigen binding domain and / or nanobody comprises the amino acid sequence of SEQ ID NO:53 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:53.

[0103] Embodiment 47 is the chimeric polypeptide of any one of embodiments 1 to 46, further comprising a signal peptide.202666783.1 - 34 -

[0104] Embodiment 48 is the chimeric polypeptide of embodiment 47, wherein the signal peptide comprises SEQ ID NO:122 or 123 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:122 or 123.

[0105] Embodiment 49 is the chimeric polypeptide of any one of embodiments 1 to 48, wherein the transmembrane domain is an α or β chain of the T cell receptor or a transmembrane domain from CD28, CD3-epsilon, CD45, CD4, CD5, CD8-α, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD123, CD134, CD137 or CD154.

[0106] Embodiment 50 is the chimeric polypeptide of embodiment 49, wherein the transmembrane domain is a CD28 transmembrane domain.

[0107] Embodiment 51 is the chimeric polypeptide of any one of embodiments 1 to 50, wherein the transmembrane domain comprises SEQ ID NO:130 or 131 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:130 or 131.

[0108] Embodiment 52 is the chimeric polypeptide of any one of embodiments 1 to 51, wherein the intracellular signaling domain comprises a primary signaling domain.

[0109] Embodiment 53 is the chimeric polypeptide of embodiment 52, wherein the primary signaling domain is a CD3ζ (zeta) signaling domain.

[0110] Embodiment 54 is the chimeric polypeptide of embodiment 52 or embodiment 53, wherein the primary signaling domain comprises SEQ ID NO:134 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:134.

[0111] Embodiment 55 is the chimeric polypeptide of any one of embodiments 1 to 54, wherein the intracellular signaling domain further comprises a costimulatory domain.

[0112] Embodiment 56 is the chimeric polypeptide of embodiment 55, wherein the costimulatory domain comprises a signaling domain from 4-1BB (CD137), CD28, IL-15Rα, OX40, CD2, CD27, CDS, ICAM-1, LFA-1 (CD11a / CD18), or ICOS (CD278).

[0113] Embodiment 57 is the chimeric polypeptide of embodiment 56, wherein the costimulatory domain comprises a CD28 or 4-1BB signaling domain.

[0114] Embodiment 58 is the chimeric polypeptide of any of embodiments 55 to 57, wherein the costimulatory domain comprises SEQ ID NO:132 or 133 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:132 or 133.

[0115] Embodiment 59 is the chimeric polypeptide of any of embodiments 1 to 58, wherein the antigen binding domain and the transmembrane domain are separated by a hinge region.

[0116] Embodiment 60 is the chimeric polypeptide of embodiment 59, wherein the hinge region comprises an IgG4 hinge, a CD8α hinge, an IgG1 hinge, or a CD34 hinge.202666783.1 - 35 -

[0117] Embodiment 61 is the chimeric polypeptide of embodiment 59 or embodiment 60, wherein the hinge region is between 8 and 300 amino acids in length.

[0118] Embodiment 62 is the chimeric polypeptide of any one of embodiments 59 to 61, wherein the hinge region comprises one of SEQ ID NOS:126-129 or an amino acid sequence having at least 80% sequence identity to one of SEQ ID NOS:126-129.

[0119] Embodiment 63 is a polypeptide comprising the amino acid sequence of any one of SEQ ID NOS:138-207 or an amino acid sequence having at least 80% sequence identity to the amino acid sequences of SEQ ID NOS:138-207.

[0120] Embodiment 64 is a nucleic acid comprising a sequence encoding a chimeric polypeptide of any of embodiments 1 to 62 or the polypeptide of embodiment 63.

[0121] Embodiment 65 is the nucleic acid of embodiment 64, wherein the nucleic acid is DNA or is RNA.

[0122] Embodiment 66 is the nucleic acid of embodiment 64 or embodiment 65, wherein the nucleic acid comprises a promoter that directs the expression of the chimeric polypeptide.

[0123] Embodiment 67 is the nucleic acid of embodiment 66, wherein the promoter is constitutive.

[0124] Embodiment 68 is the nucleic acid of embodiment 66, wherein the promoter is conditional.

[0125] Embodiment 69 is the nucleic acid of embodiment 68, wherein the promoter responds positively to at least one cytokine or to T-cell activation.

[0126] Embodiment 70 is the nucleic acid of embodiment 69 where the at least one cytokine is TGF- β, IL-6, TNF α, IFN-γ, IL-1β, IL-2, IL-8, or IL-10 or the promoter responds positively to NFAT or NF-κB.

[0127] Embodiment 71 is the nucleic acid of any one of embodiments 64 to 70, wherein the nucleic acid is an expression construct.

[0128] Embodiment 72 is the nucleic acid of embodiment 71, wherein the expression construct is a viral vector.

[0129] Embodiment 73 is the nucleic acid of embodiment 72, wherein the viral vector comprises a retroviral vector a vector derived from a retrovirus.

[0130] Embodiment 74 is the nucleic acid of embodiment 73, wherein the viral vector is a lentiviral vector or a vector derived from a lentivirus.

[0131] Embodiment 75 is a lentivirus vector comprising a sequence encoding the chimeric polypeptide of any one of embodiments 1 to 62 or the polypeptide of embodiment 63.202666783.1 - 36 -

[0132] Embodiment 76 is a cell comprising the nucleic acid of any of embodiments 64 to 75.

[0133] Embodiment 77 is the cell of embodiment 76, wherein the viral vector has integrated into the cell’s genome.

[0134] Embodiment 78 is a cell expressing a chimeric polypeptide of any of embodiments 1 to 62 or the polypeptide of embodiment 63.

[0135] Embodiment 79 is the cell of any of embodiments 76 to 78, wherein the cell is a T cell, a natural killer (NK) cell, a natural killer T cell (NKT), an invariant natural killer T cell (iNKT), stem cell, lymphoid progenitor cell, peripheral blood mononuclear cell (PBMC), bone marrow cell, fetal liver cell, embryonic stem cell, cord blood cell, induced pluripotent stem cell (iPS cell).

[0136] Embodiment 80 is the cell of embodiment 79, wherein the cell is a T cell or an NK cell.

[0137] Embodiment 81 is the cell of embodiment 80, wherein the T cell comprises a naïve memory T cell.

[0138] Embodiment 82 is the cell of embodiment 81, wherein the naïve memory T cell comprises a CD4+ or CD8+ T cell.

[0139] Embodiment 83 is the cell of any one of embodiments 80 to 82, wherein the T cell comprises a T cell from a population of CD14 depleted, CD25 depleted, and / or CD62L enriched PBMCs.

[0140] Embodiment 84 is a population of cell comprising any of the cells of embodiments 76 to 82.

[0141] Embodiment 85 is the population of cells of embodiment 84, wherein the population comprises 103-108cells.

[0142] Embodiment 86 is a composition comprising the population of cells of embodiment 84 or embodiment 85, wherein the composition is a pharmaceutically acceptable formulation.

[0143] Embodiment 87 is a method of making a cell comprising introducing into a cell the nucleic acid of any of embodiments 64 to 74.

[0144] Embodiment 88 is the method of embodiment 87, wherein the cell is infected with a virus encoding the CAR.

[0145] Embodiment 89 is the method of embodiment 88, wherein the virus comprises lentivirus or a lentiviral-derived virus or vector.

[0146] Embodiment 90 is the method of any one of embodiments 87 to 89, wherein the cell is a T cell, a natural killer (NK) cell, a natural killer T cell (NKT), an invariant natural killer T202666783.1 - 37 -cell (iNKT), stem cell, lymphoid progenitor cell, peripheral blood mononuclear cell (PBMC), bone marrow cell, fetal liver cell, embryonic stem cell, cord blood cell, induced pluripotent stem cell (iPS cell).

[0147] Embodiment 91 is the method of embodiment 90, wherein the cell is a T cell or an NK cell.

[0148] Embodiment 92 is the method of embodiment 91, wherein the T cell comprises a naïve memory T cell.

[0149] Embodiment 93 is the method of embodiment 92, wherein the naïve memory T cell comprises a CD4+ or CD8+ T cell.

[0150] Embodiment 94 is the method of any one of embodiments 87 to 93, wherein the T cell comprises a T cell from a population of CD14 depleted, CD25 depleted, and / or CD62L enriched PBMCs.

[0151] Embodiment 95 is the method of any one of embodiments 90 to 94, wherein the cell is not yet a T cell or NK cell, the method further comprising culturing the cell under conditions that promote the differentiation of the cell into a T cell or an NK cell.

[0152] Embodiment 96 is the method of any of embodiments 87 to 95, further comprising culturing the cell under conditions to expand the cell before and or after introducing the nucleic acid into the cell.

[0153] Embodiment 97 is the method of embodiment 96, wherein the cell is cultured with serum-free medium.

[0154] Embodiment 98 is a method of treating a patient with cancer comprising administering to the patient an effective amount of the composition of embodiment 86.

[0155] Embodiment 99 is the method of embodiment 98, wherein the patient has and / or has been determined to have a MSLN+ cancer.

[0156] Embodiment 100 is the method of embodiment 98 or embodiment 99, wherein the cancer comprise ovarian cancer, stomach cancer, colorectal cancer, endometrial cancer, uterine cancer, acute myeloid leukemia, pancreatic cancer, lung cancer, or mesothelioma.

[0157] Embodiment 101 is the method of any one of embodiments 98 to 100, further comprising administering an additional therapy to the patient.

[0158] Embodiment 102 is the method of any one of embodiments 98 to 101, further comprising administering an additional therapy to the patient.

[0159] Embodiment 103 is the method of embodiment 102, wherein the additional therapy comprises an immunotherapy, anti-angiogenic therapy, chemotherapy, surgery, radiotherapy, neoantigen therapy, or vaccination.202666783.1 - 38 -

[0160] Embodiment 104 is the method of embodiment 103, wherein the additional therapy comprises immunotherapy and wherein the immunotherapy comprises immune checkpoint inhibitor therapy or bispecific T cell engagers.

[0161] Embodiment 105 is the method of any one of embodiments 98 to 104, wherein the cells are autologous.

[0162] Embodiment 106 is a nucleic acid encoding: (i) a chimeric antigen receptor (CAR) or engineered T-cell receptor (TCR); and (ii) an IL-12 polypeptide; and / or (iii) an IL-18 polypeptide.

[0163] Embodiment 107 is a nucleic acid encoding: (i) a chimeric antigen receptor (CAR) or engineered T-cell receptor (TCR); and (ii) an IL-12 polypeptide; (iii) an IFN-β polypeptide; and / or (iv) a CXCL9 polypeptide.

[0164] Embodiment 108 is the nucleic acid of embodiment 106 or embodiment 107, wherein the nucleic acid is an expression construct.

[0165] Embodiment 109 is the nucleic acid of any one of embodiments 106-108, wherein the expression construct is a plasmid or viral vector.

[0166] Embodiment 110 is the nucleic acid of embodiment 109, wherein the viral vector is a vector derived from a retrovirus or a vector derived from a lentivirus.

[0167] Embodiment 111 is the nucleic acid of any one of embodiments 106 to 109, wherein the IL-12 polypeptide comprises a p40 and / or p35 subunit.

[0168] Embodiment 112 is the nucleic acid of any one of embodiments 106 to 111, wherein the IL-12 polypeptide comprises a single-chain IL-12 polypeptide.

[0169] Embodiment 113 is the nucleic acid of embodiment 112, wherein the p40 subunit is amino-proximal to the p35 subunit.

[0170] Embodiment 114 is the nucleic acid of embodiment 112, wherein the p35 subunit is amino-proximal to the p40 subunit.

[0171] Embodiment 115 is the nucleic acid of any one of embodiments 112 to 114, wherein the nucleic acid encodes for a linker between the p40 and p35 subunit.

[0172] Embodiment 116 is the nucleic acid of any one of embodiments 106 to 115, wherein the nucleic acid further encodes for a suicide gene product.

[0173] Embodiment 117 is the nucleic acid of embodiment 116, wherein suicide gene product is herpes simplex virus thymidine kinase (HSV-TK), purine nucleoside phosphorylase (PNP), cytosine deaminase (CD), carboxypetidase G2, cytochrome P450, linamarase, β- lactamase, nitroreductase (NTR), carboxypeptidase A, split Cas9, truncated EGFR, or inducible caspase 9.202666783.1 - 39 -

[0174] Embodiment 118 is the nucleic acid of any one of embodiments 106 to 117, wherein the nucleic acid comprises one or more promoter(s) that direct the expression of the suicide gene product, CAR, IL-12 polypeptide, IFN-β polypeptide, CXCL9 polypeptide, and / or IL-18 polypeptide.

[0175] Embodiment 119 is the nucleic acid of embodiment 118, wherein the promoter is constitutive.

[0176] Embodiment 120 is the nucleic acid of embodiment 118, wherein the promoter is conditional.

[0177] Embodiment 121 is the nucleic acid of embodiment 120, wherein the promoter responds positively to at least one cytokine or to T-cell activation.

[0178] Embodiment 122 is the nucleic acid of embodiment 121, where the at least one cytokine is TGF- β, IL-6, TNF α, IFN-γ, IL-1β, IL-2, IL-8, or IL-10 or the promoter responds positively to NFAT or NF-κB.

[0179] Embodiment 123 is the nucleic acid of any one of embodiments 106 to 122, wherein the IL-12, IFN-β, CXCL9, and / or IL-18 polypeptide is derived from a human IL-12, human IFN-β, human CXCL9, and / or human IL-18 polypeptide.

[0180] Embodiment 124 is the nucleic acid of any one of embodiments 106 to 123, wherein the IL-18 polypeptide comprises a decoy resistant IL-18 polypeptide.

[0181] Embodiment 125 is the nucleic acid of any one of embodiments 111 to 124, wherein the p40 polypeptide comprises the amino acid sequence of SEQ ID NO:236 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:236.

[0182] Embodiment 126 is the nucleic acid of any one of embodiments 111 to 125, wherein the p35 polypeptide comprises the amino acid sequence of SEQ ID NO:237 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:237.

[0183] Embodiment 127 is the nucleic acid of any one of embodiments 106 to 126, wherein the IL-12 polypeptide comprises the amino acid sequence of SEQ ID NO:230 or 234 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:230 or 234.

[0184] Embodiment 128 is the nucleic acid of any one of embodiments 106 to 127, wherein the IL-18 polypeptide comprises the amino acid sequence of SEQ ID NO:229, 232, or 238-240 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:229, 232, or 238-240.

[0185] Embodiment 129 is the nucleic acid of any one of embodiments 107 to 128, wherein the IFN-β polypeptide comprises the amino acid sequence of SEQ ID NO:253 or 255 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:253 or 255.202666783.1 - 40 -

[0186] Embodiment 130 is the nucleic acid of any one of embodiments 107 to 128, wherein the CXCL9 polypeptide comprises the amino acid sequence of SEQ ID NO:257 or 259 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:257 or 259.

[0187] Embodiment 131 is the nucleic acid of any one of embodiments 106 to 130, wherein the nucleic acid encodes for a signal sequence at the amino-terminus of the IL-12, IFN-β, CXCL9 and / or IL-18 polypeptide.

[0188] Embodiment 132 is the nucleic acid of embodiment 131, wherein the signal sequence comprises SEQ ID NO:122, 123, 235, 241, 254, 256, 258, and / or 260.

[0189] Embodiment 133 is the nucleic acid of any one of embodiments 106 to 132, wherein the IL-12 polypeptide and the IL-18 polypeptide are expressed from one nucleic acid.

[0190] Embodiment 134 is the nucleic acid of embodiment 133, wherein the nucleic acid expresses a polypeptide comprising the IL-12 polypeptide and the IL-18 polypeptide.

[0191] Embodiment 135 is the nucleic acid of any one of embodiments 107 to 132, wherein the IL-12 polypeptide, IFN-β polypeptide, and the CXCL9 polypeptide are expressed from one nucleic acid.

[0192] 136 is the nucleic acid of embodiment 135, wherein the nucleic acid expresses a polypeptide comprising the IL-12, IFN-β, and CXCL9 polypeptide.

[0193] Embodiment 137 is the nucleic acid of any one of embodiments 133 to 136, wherein the nucleic acid encodes for a self-cleaving peptide or internal ribosome entry sequence (IRES), wherein the self-cleaving peptide or IRES is between the IL-12, IL-18, CXCL9, and / or IFN-β polypeptides.

[0194] Embodiment 138 is the nucleic acid of embodiment 137, wherein the self-cleaving polypeptide comprises SEQ ID NO:135 or 136.

[0195] Embodiment 139 is the nucleic acid of any one of embodiments 106 to 138, wherein the nucleic acid encodes for a chimeric antigen receptor (CAR), wherein the CAR comprises: (a) an antigen binding domain; (b) a transmembrane domain; and (c) an intracellular signaling domain.

[0196] Embodiment 140 is the nucleic acid of embodiment 139, wherein the antigen binding domain comprises an anti-TYRP-1, anti-IL-13Rα2, anti-TGF-β, anti-CLDN6, anti- GPC3, or anti-MSLN antigen binding domain.

[0197] Embodiment 141 is the nucleic acid of embodiment 140, wherein the antigen binding domain comprises an anti-IL-13Rα2 comprising an IL-13 mutein polypeptide.202666783.1 - 41 -

[0198] Embodiment 142 is the nucleic acid of embodiment 140, wherein the IL-13 mutein polypeptide comprises the amino acid sequence of SEQ ID NO:115 or 233, or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:115 or 233.

[0199] Embodiment 143 is the nucleic acid of embodiment 140, wherein the antigen binding domain comprises anti-MSLN.

[0200] Embodiment 144 is the nucleic acid of embodiment 143, wherein the antigen binding domain comprises a nanobody, wherein the nanobody comprises a complementarity determining region (CDR) 1, CDR2, and CDR3 having at least 80% sequence identity to the CDR1, CDR2, CDR3 from the nanobody of SEQ ID NO:57.

[0201] Embodiment 145 is the nucleic acid of embodiment 144, wherein the nanobody comprises a CDR1, CDR2, and CDR3 having the amino acid sequence of the CDR1, CDR2, and CDR3, respectively, from the nanobody of SEQ ID NO:57.

[0202] Embodiment 146 is the nucleic acid of embodiment 144 or embodiment 145, wherein the CDR1, CDR2, and CDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:58-60, respectively.

[0203] Embodiment 147 is the nucleic acid of any one of embodiments 144 to 146, wherein the CDR1, CDR2, and CDR3 each comprise the amino acid sequence of SEQ ID NOS:58-60, respectively.

[0204] Embodiment 148 is the nucleic acid of any one of embodiments 144 to 147, wherein the nanobody comprises a framework region (FR) 1, FR2, FR3, and FR4, wherein the FR1, FR2, FR3, and FR4 comprises an amino acid sequence with at least 80% sequence identity to an FR1, FR2, FR3, and FR4, respectively, of a nanobody comprising the amino acid sequence of SEQ ID NO:57.

[0205] Embodiment 149 is the nucleic acid of embodiment 148, wherein the FR1, FR2, FR3, and FR4 comprises the amino acid sequence of SEQ ID NOS:225-228, respectively or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:225-228, respectively.

[0206] Embodiment 150 is the nucleic acid of embodiment 143, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:36 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1,202666783.1 - 42 -LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:37.

[0207] Embodiment 151 is the nucleic acid of embodiment 150, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:36 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:37.

[0208] Embodiment 152 is the nucleic acid of embodiment 150 or embodiment 151, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:38-43, respectively.

[0209] Embodiment 153 is the nucleic acid of any one of embodiments 150 to 152, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:38-43, respectively.

[0210] Embodiment 154 is the nucleic acid of any one of embodiments 150 to 153, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:36 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:37.

[0211] Embodiment 155 is the nucleic acid of embodiment 154, wherein the VH comprises the amino acid sequence of SEQ ID NO:36 and / or the VL comprises the amino acid sequence of SEQ ID NO:37.

[0212] Embodiment 156 is the nucleic acid of any one of embodiments 150 to 155, wherein the VH comprises a heavy chain framework region (HFR) 1, HFR2, HFR3, and HFR4 and the VL comprises a light chain framework region (LFR) 1, LFR2, LFR3, and LFR4, and wherein the HFR1, HFR2, HFR3, and HFR4 comprises an amino acid sequence with at least 80% sequence identity to an HFR1, HFR2, HFR3, and HFR4, respectively, of a VH comprising the amino acid sequence of SEQ ID NO:36, and the LFR1, LFR2, LFR3, and LFR4 comprises an amino acid sequence with at least 80% sequence identity to the LFR1, LFR2, LFR3, and LFR4, respectively, of a VL comprising the amino acid sequence of SEQ ID NO:37.

[0213] Embodiment 157 is the nucleic acid of any one of embodiments 150 to 156, wherein the HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4 comprises the amino acid sequence of SEQ ID NOS:208-215, respectively or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:208-215, respectively.202666783.1 - 43 -

[0214] Embodiment 158 is the nucleic acid of embodiment 143, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:45 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:46.

[0215] Embodiment 159 is the nucleic acid of embodiment 158, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:45 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:46.

[0216] Embodiment 160 is the nucleic acid of embodiment 158 or embodiment 159, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:47-52, respectively.

[0217] Embodiment 161 is the nucleic acid of any one of embodiments 158 to 160, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:47-52, respectively.

[0218] Embodiment 162 is the nucleic acid of any one of embodiments 158 to 161, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:45 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:46.

[0219] Embodiment 163 is the nucleic acid of embodiment 162, wherein the VH comprises the amino acid sequence of SEQ ID NO:45 and / or the VL comprises the amino acid sequence of SEQ ID NO:46.

[0220] Embodiment 164 is the nucleic acid of any one of embodiments 158 to 163, wherein the VH comprises a heavy chain framework region (HFR) 1, HFR2, HFR3, and HFR4 and the VL comprises a light chain framework region (LFR) 1, LFR2, LFR3, and LFR4, and wherein the HFR1, HFR2, HFR3, and HFR4 comprises an amino acid sequence with at least 80% sequence identity to an HFR1, HFR2, HFR3, and HFR4, respectively, of a VH comprising the amino acid sequence of SEQ ID NO:45, and the LFR1, LFR2, LFR3, and LFR4 comprises an202666783.1 - 44 -amino acid sequence with at least 80% sequence identity to the LFR1, LFR2, LFR3, and LFR4, respectively, of a VL comprising the amino acid sequence of SEQ ID NO:46.

[0221] Embodiment 165 is the nucleic acid of any one of embodiments 158 to 164, wherein the HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4 comprises the amino acid sequence of SEQ ID NOS:216, 209, 217, 211, 218, 219, 220, and 215, respectively or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:216, 209, 217, 211, 218, 219, 220, and 215, respectively.

[0222] Embodiment 166 is the nucleic acid of embodiment 143, wherein the antigen binding domain comprises a nanobody, wherein the nanobody comprises a complementarity determining region (CDR) 1, CDR2, and CDR3 having at least 80% sequence identity to the CDR1, CDR2, CDR3 from the nanobody of SEQ ID NO:53.

[0223] Embodiment 167 is the nucleic acid of embodiment 166, wherein the nanobody comprises a CDR1, CDR2, and CDR3 having the amino acid sequence of the CDR1, CDR2, and CDR3, respectively, from the nanobody of SEQ ID NO:53.

[0224] Embodiment 168 is the nucleic acid of embodiment 166 or embodiment 167, wherein the CDR1, CDR2, and CDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:54-56, respectively.

[0225] Embodiment 169 is the nucleic acid of any one of embodiments 166 to 168, wherein the CDR1, CDR2, and CDR3 each comprise the amino acid sequence of SEQ ID NOS:54-56, respectively.

[0226] Embodiment 170 is the nucleic acid of any one of embodiments 166 to 169, wherein the nanobody comprises a framework region (FR) 1, FR2, FR3, and FR4, wherein the FR1, FR2, FR3, and FR4 comprises an amino acid sequence with at least 80% sequence identity to an FR1, FR2, FR3, and FR4, respectively, of a nanobody comprising the amino acid sequence of SEQ ID NO:53.

[0227] Embodiment 171 is the nucleic acid of any one of embodiments 166 to 170, wherein the FR1, FR2, FR3, and FR4 comprises the amino acid sequence of SEQ ID NOS:221-224, respectively or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:221-224, respectively.

[0228] Embodiment 172 is the nucleic acid of embodiment 140, wherein the antigen binding domain comprises anti-TYRP-1.

[0229] Embodiment 173 is the nucleic acid of embodiment 172, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1,202666783.1 - 45 -HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:62 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:63.

[0230] Embodiment 174 is the nucleic acid of embodiment 173, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:62 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:63.

[0231] Embodiment 175 is the nucleic acid of embodiment 173 or embodiment 174, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:64-69, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:64-69, respectively.

[0232] Embodiment 176 is the nucleic acid of any one of embodiments 173 to 175, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:62 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:63, or wherein the VH comprises the amino acid sequence of SEQ ID NO:62 and / or the VL comprises the amino acid sequence of SEQ ID NO:63.

[0233] Embodiment 177 is the nucleic acid of embodiment 140, wherein the antigen binding domain comprises anti-TGF-β.

[0234] Embodiment 178 is the nucleic acid of embodiment 177, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:71 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:72.

[0235] Embodiment 179 is the nucleic acid of embodiment 178, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:71 and202666783.1 - 46 -wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:72.

[0236] Embodiment 180 is the nucleic acid of embodiment 178 or embodiment 179, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:73-78, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:73-78, respectively.

[0237] Embodiment 181 is the nucleic acid of any one of embodiments 178 to 180, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:71 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:72, or wherein the VH comprises the amino acid sequence of SEQ ID NO:71 and / or the VL comprises the amino acid sequence of SEQ ID NO:72.

[0238] Embodiment 182 is the nucleic acid of embodiment 177, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:80 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:81.

[0239] Embodiment 183 is the nucleic acid of embodiment 182, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:80 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:81.

[0240] Embodiment 184 is the nucleic acid of embodiment 182 or embodiment 183, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:82-87, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:82-87, respectively.

[0241] Embodiment 185 is the nucleic acid of any one of embodiments 182 to 184, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID202666783.1 - 47 -NO:80 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:81, or wherein the VH comprises the amino acid sequence of SEQ ID NO:80 and / or the VL comprises the amino acid sequence of SEQ ID NO:81.

[0242] Embodiment 186 is the nucleic acid of embodiment 177, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:89 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:90.

[0243] Embodiment 187 is the nucleic acid of embodiment 186, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:89 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:90.

[0244] Embodiment 188 is the nucleic acid of embodiment 186 or embodiment 187, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:91-96, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:91-96, respectively.

[0245] Embodiment 189 is the nucleic acid of any one of embodiments 186 to 188, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:89 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:90, or wherein the VH comprises the amino acid sequence of SEQ ID NO:89 and / or the VL comprises the amino acid sequence of SEQ ID NO:90.

[0246] Embodiment 190 is the nucleic acid of embodiment 140, wherein the antigen binding domain comprises anti-CLDN6.

[0247] Embodiment 191 is the nucleic acid of embodiment 190, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:98 and wherein the light chain202666783.1 - 48 -variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:99.

[0248] Embodiment 192 is the nucleic acid of embodiment 191, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:98 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:99.

[0249] Embodiment 193 is the nucleic acid of embodiment 191 or embodiment 192, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:100-103, the sequence TSN, and SEQ ID NO:105, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:100- 103, the sequence TSN, and SEQ ID NO:105, respectively.

[0250] Embodiment 194 is the nucleic acid of any one of embodiments 191 to 193, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:98 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:99, or wherein the VH comprises the amino acid sequence of SEQ ID NO:98 and / or the VL comprises the amino acid sequence of SEQ ID NO:99.

[0251] Embodiment 195 is the nucleic acid of embodiment 140, wherein the antigen binding domain comprises anti-GPC3.

[0252] Embodiment 196 is the nucleic acid of embodiment 195, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:107 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:108.

[0253] Embodiment 197 is the nucleic acid of embodiment 191, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:107 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of202666783.1 - 49 -the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:108.

[0254] Embodiment 198 is the nucleic acid of embodiment 191 or embodiment 192, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:109-114, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:109-114, respectively.

[0255] Embodiment 199 is the nucleic acid of any one of embodiments 191 to 193, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:107 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:108, or wherein the VH comprises the amino acid sequence of SEQ ID NO:107 and / or the VL comprises the amino acid sequence of SEQ ID NO:108.

[0256] Embodiment 200 is the nucleic acid of any one of embodiments 150 to 165 or embodiments 173 to 199, wherein the VH is amino proximal to the VL.

[0257] Embodiment 201 is the nucleic acid of any one of embodiments 150 to 165 or embodiments 173 to 199, wherein the VL is amino proximal to the VH.

[0258] Embodiment 202 is the nucleic acid of any one of embodiments 150 to 165 or embodiments 173 to 201, wherein the antigen binding domain comprises a linker between the VH and VL.

[0259] Embodiment 203 is the nucleic acid of embodiment 202, wherein the linker comprises a glycine serine linker.

[0260] Embodiment 204 is the nucleic acid of embodiment 203, wherein the linker comprises (G4S)n, wherein n is an integer selected from 1-10.

[0261] Embodiment 205 is the nucleic acid of embodiment 204, wherein the linker comprises or consists of SEQ ID NO:119 or 121.

[0262] Embodiment 206 is the nucleic acid of embodiment 202, wherein the linker comprises SEQ ID NO:120.

[0263] Embodiment 207 is the nucleic acid of any one of embodiments 150 to 206, wherein the antigen binding domain comprises the amino acid sequence of one of SEQ ID NOS:35, 44, 53, 57, 61, 70, 79, 88, 97, or 106 or an amino acid sequence having at least 80% sequence identity to one of SEQ ID NOS:35, 44, 53, 57, 61, 70, 79, 88, 97, or 106.

[0264] Embodiment 208 is the nucleic acid of any one of embodiments 106 to 207, wherein the nucleic acid is DNA or is RNA.202666783.1 - 50 -

[0265] Embodiment 209 is the nucleic acid of any one of embodiments 106 to 207, further comprising a signal peptide.

[0266] Embodiment 210 is the nucleic acid of embodiment 209, wherein the signal peptide comprises SEQ ID NO:122 or 123 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:122 or 123.

[0267] Embodiment 211 is the nucleic acid of any one of embodiments 139 to 210, wherein the transmembrane domain is an α or β chain of the T cell receptor or a transmembrane domain from CD28, CD3-epsilon, CD45, CD4, CD5, CD8-α, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD123, CD134, CD137 or CD154.

[0268] Embodiment 212 is the nucleic acid of embodiment 211, wherein the transmembrane domain is a CD28 transmembrane domain.

[0269] Embodiment 213 is the nucleic acid of any one of embodiments 139 to 212, wherein the transmembrane domain comprises SEQ ID NO:130 or 131 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:130 or 131.

[0270] Embodiment 214 is the nucleic acid of any one of embodiments 139 to 213, wherein the intracellular signaling domain comprises a primary signaling domain.

[0271] Embodiment 215 is the nucleic acid of embodiment 214, wherein the primary signaling domain is a CD3ζ (zeta) signaling domain.

[0272] Embodiment 216 is the nucleic acid of embodiment 214 or embodiment 215, wherein the primary signaling domain comprises SEQ ID NO:134 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:134.

[0273] Embodiment 217 is the nucleic acid of any one of embodiments 139 to 216, wherein the intracellular signaling domain further comprises a costimulatory domain.

[0274] Embodiment 218 is the nucleic acid of embodiment 217, wherein the costimulatory domain comprises a signaling domain from 4-1BB (CD137), CD28, IL-15Rα, OX40, CD2, CD27, CDS, ICAM-1, LFA-1 (CD11a / CD18), or ICOS (CD278).

[0275] Embodiment 219 is the nucleic acid of embodiment 218, wherein the costimulatory domain comprises a CD28 or 4-1BB signaling domain.

[0276] Embodiment 220 is the nucleic acid of any of embodiments 217 to 219, wherein the costimulatory domain comprises SEQ ID NO:132 or 133 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:132 or 133.

[0277] Embodiment 221 is the nucleic acid of any of embodiments 139 to 220, wherein the antigen binding domain and the transmembrane domain are separated by a hinge region.202666783.1 - 51 -

[0278] Embodiment 222 is the nucleic acid of embodiment 221, wherein the hinge region comprises an IgG4 hinge, a CD8α hinge, an IgG1 hinge, or a CD34 hinge.

[0279] Embodiment 223 is the nucleic acid of embodiment 221 or embodiment 222, wherein the hinge region is between 8 and 300 amino acids in length.

[0280] Embodiment 224 is the nucleic acid of any one of embodiments 221 to 223, wherein the hinge region comprises one of SEQ ID NOS:126-129 or an amino acid sequence having at least 80% sequence identity to one of SEQ ID NOS:126-129.

[0281] Embodiment 225 is a nucleic acid encoding a polypeptide comprising the amino acid sequence of one of SEQ ID NOS:138-207 or an amino acid sequence having at least 80% sequence identity to one of SEQ ID NOS:138-207.

[0282] Embodiment 226 is a polypeptide expressed from the nucleic acid of any one of embodiments 106 to 225.

[0283] Embodiment 227 is a cell or population of cells comprising the nucleic acid of any one of embodiments 106 to 225 or the polypeptide of embodiment 226.

[0284] Embodiment 228 is the cell or cells of embodiment 227, wherein the cell or cells express a polypeptide encoded by the nucleic acid.

[0285] Embodiment 229 is a cell or population of cells comprising a heterologous nucleic acid encoding an IL-12 polypeptide and a heterologous nucleic acid encoding an IL-18 polypeptide.

[0286] Embodiment 230 is a cell or population of cells comprising a heterologous nucleic acid encoding an IL-12 polypeptide, a heterologous nucleic acid encoding an IFN-β polypeptide and a heterologous nucleic acid encoding a CXCL9 polypeptide.

[0287] Embodiment 231 is the cell of embodiment 229 or embodiment 230, wherein the IL-12 polypeptide comprises a p40 and / or p35 subunit.

[0288] Embodiment 232 is the cell of any one of embodiments 229 to 231, wherein the IL- 12 polypeptide comprises a single-chain IL-12 polypeptide.

[0289] Embodiment 233 is the cell of embodiment 232, wherein the p40 subunit is amino- proximal to the p35 subunit.

[0290] Embodiment 234 is the cell of embodiment 232, wherein the p35 subunit is amino- proximal to the p40 subunit.

[0291] Embodiment 235 is the cell of any one of embodiments 232 to 234, wherein the IL- 12 polypeptide comprises a linker between the p40 and p35 subunit.

[0292] Embodiment 236 is the cell or cells of embodiment 235, wherein the nucleic acid encoding the IL-12, IFN-β, CXCL9 and / or IL-18 polypeptide is an expression construct.202666783.1 - 52 -

[0293] Embodiment 237 is the cell or cells of embodiment 236, wherein the expression construct is a plasmid or viral vector.

[0294] Embodiment 238 is the cell or cells of embodiment 237, wherein the viral vector is a vector derived from a retrovirus or a vector derived from a lentivirus.

[0295] Embodiment 239 is the cell or cells of any one of embodiments 227 to 238, wherein the cell comprises a nucleic acid encoding for a suicide gene product.

[0296] Embodiment 240 is the cell or cells of embodiment 239, wherein suicide gene product is herpes simplex virus thymidine kinase (HSV-TK), purine nucleoside phosphorylase (PNP), cytosine deaminase (CD), carboxypetidase G2, cytochrome P450, linamarase, β- lactamase, nitroreductase (NTR), carboxypeptidase A, split Cas9, truncated EGFR, or inducible caspase 9.

[0297] Embodiment 241 is the cell or cells of any one of embodiments 229 to 240, wherein the nucleic acid comprises one or more promoter(s) that direct the expression of the suicide gene product, IL-12, IFN-β, CXCL9, and / or IL-18 polypeptide.

[0298] Embodiment 242 is the cell or cells of embodiment 241, wherein the promoter is constitutive.

[0299] Embodiment 243 is the cell or cells of embodiment 241, wherein the promoter is conditional.

[0300] Embodiment 244 is the cell or cells of embodiment 243, wherein the promoter responds positively to at least one cytokine or to T-cell activation.

[0301] Embodiment 245 is the cell or cells of embodiment 244, where the at least one cytokine is TGF- β, IL-6, TNF α, IFN-γ, IL-1β, IL-2, IL-8, or IL-10 or the promoter responds positively to NFAT or NF-κB.

[0302] Embodiment 246 is the cell or cells of any one of embodiments 229 to 245, wherein the IL-12, IFN- β, CXCL9, and / or IL-18 polypeptide is derived from a human IL-12, human IFN- β, human CXCL9, and / or human IL-18 polypeptide.

[0303] Embodiment 247 is the cell or cells of any one of embodiments 229 to 246, wherein the IL-18 polypeptide comprises a decoy resistant IL-18 polypeptide.

[0304] Embodiment 248 is the cell or cells of any one of embodiments 231 to 247, wherein the p40 polypeptide comprises the amino acid sequence of SEQ ID NO:236 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:236.

[0305] Embodiment 249 is the cell or cells of any one of embodiments 231 to 248, wherein the p35 polypeptide comprises the amino acid sequence of SEQ ID NO:237 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:237.202666783.1 - 53 -

[0306] Embodiment 250 is the cell or cells of any one of embodiments 229 to 249, wherein the IL-12 polypeptide comprises the amino acid sequence of SEQ ID NO:230 or 234 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:230 or 234.

[0307] Embodiment 251 is the cell or cells of any one of embodiments 229 to 250, wherein the IL-18 polypeptide comprises the amino acid sequence of SEQ ID NO:229, 232, or 238-240 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:229, 232, or 238-240.

[0308] Embodiment 252 is the cell or cells of any one of embodiments 229-251, wherein the IFN-β polypeptide comprises the amino acid sequence of SEQ ID NO:253 or 255 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:253 or 255.

[0309] Embodiment 253 is the cell or cells of any one of embodiments 229 to 252, wherein the CXCL9 polypeptide comprises the amino acid sequence of SEQ ID NO:257 or 259 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:257 or 259.

[0310] Embodiment 254 is the cell or cells of any one of embodiments 229 to 253, wherein the nucleic acid encodes for a signal sequence at the amino-terminus of the IL-12, IFN-β, CXCL9 and / or IL-18 polypeptide.

[0311] Embodiment 255 is the cell or cells of any one of embodiments 229 to 254, wherein the signal sequence comprises SEQ ID NO:122, 123, 235, 241, 254, 256, 258, and / or 260.

[0312] Embodiment 256 is the cell or cells of any one of embodiments 229 to 255, wherein the IL-12 polypeptide and the IL-18 polypeptide are expressed from one nucleic acid.

[0313] Embodiment 257 is the cell or cells of embodiment 256, wherein the nucleic acid expresses a polypeptide comprising the IL-12 polypeptide and the IL-18 polypeptide.

[0314] Embodiment 258 is the cell or cells of any one of embodiments 229 to 255, wherein the IL-12 polypeptide, IFN-β polypeptide, and the CXCL9 polypeptide are expressed from one nucleic acid.

[0315] Embodiment 259 is the nucleic acid of embodiment 258, wherein the nucleic acid expresses a polypeptide comprising the IL-12, IFN-β, and CXCL9 polypeptide.

[0316] Embodiment 260 is the cell or cells of any one of embodiments 256 to 259, wherein the nucleic acid encodes for a self-cleaving peptide or internal ribosome entry sequence (IRES), wherein the self-cleaving peptide or IRES is between the IL-12, IL-18, CXCL9, and / or IFN-β polypeptides..

[0317] Embodiment 261 is the cell or cells of embodiment 260, wherein the self-cleaving polypeptide comprises SEQ ID NO:135 or 136.202666783.1 - 54 -

[0318] Embodiment 262 is the cell or cells of any one of embodiments 229 to 261, wherein the cell further comprises a nucleic acid encoding a CAR or TCR.

[0319] Embodiment 263 is the cell or cells of embodiment 262, wherein the nucleic acid encodes for a chimeric antigen receptor (CAR), wherein the CAR comprises: (a) an antigen binding domain; (b) a transmembrane domain; and (c) an intracellular signaling domain.

[0320] Embodiment 264 is the cell or cells embodiment 263, wherein the antigen binding domain comprises an anti-TYRP-1, anti-IL-13Rα2, anti-TGF-β, anti-CLDN6, anti-GPC3, or anti-MSLN antigen binding domain.

[0321] Embodiment 265 is the cell or cells of embodiment 264, wherein the antigen binding domain comprises an anti-IL-13Rα2 comprising an IL-13 mutein polypeptide.

[0322] Embodiment 266 is the cell or cells of embodiment 264, wherein the IL-13 mutein polypeptide comprises the amino acid sequence of SEQ ID NO:115 or 233, or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:115 or 233.

[0323] Embodiment 267 is the cell or cells of embodiment 264, wherein the antigen binding domain comprises anti-MSLN.

[0324] Embodiment 268 is the cell or cells of embodiment 267, wherein the antigen binding domain comprises a nanobody, wherein the nanobody comprises a complementarity determining region (CDR) 1, CDR2, and CDR3 having at least 80% sequence identity to the CDR1, CDR2, CDR3 from the nanobody of SEQ ID NO:57.

[0325] Embodiment 269 is the cell or cells of embodiment 268, wherein the nanobody comprises a CDR1, CDR2, and CDR3 having the amino acid sequence of the CDR1, CDR2, and CDR3, respectively, from the nanobody of SEQ ID NO:57.

[0326] Embodiment 270 is the cell or cells of embodiment 268 or embodiment 269, wherein the CDR1, CDR2, and CDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:58-60, respectively.

[0327] Embodiment 271 is the cell or cells of any one of embodiments 268 to 270, wherein the CDR1, CDR2, and CDR3 each comprise the amino acid sequence of SEQ ID NOS:58-60, respectively.

[0328] Embodiment 272 is the cell or cells of any one of embodiments 268 to 271, wherein the nanobody comprises a framework region (FR) 1, FR2, FR3, and FR4, wherein the FR1, FR2, FR3, and FR4 comprises an amino acid sequence with at least 80% sequence identity to an FR1, FR2, FR3, and FR4, respectively, of a nanobody comprising the amino acid sequence of SEQ ID NO:57.202666783.1 - 55 -

[0329] Embodiment 273 is the cell or cells of embodiment 272, wherein the FR1, FR2, FR3, and FR4 comprises the amino acid sequence of SEQ ID NOS:225-228, respectively or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:225-228, respectively.

[0330] Embodiment 274 is the cell or cells of embodiment 267, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:36 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:37.

[0331] Embodiment 275 is the cell or cells of embodiment 274, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:36 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:37.

[0332] Embodiment 276 is the cell or cells of embodiment 274 or embodiment 275, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:38-43, respectively.

[0333] Embodiment 277 is the cell or cells of any one of embodiments 274 to 276, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:38-43, respectively.

[0334] Embodiment 278 is the cell or cells of any one of embodiments 274 to 277, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:36 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:37.

[0335] Embodiment 279 is the cell or cells of embodiment 278, wherein the VH comprises the amino acid sequence of SEQ ID NO:36 and / or the VL comprises the amino acid sequence of SEQ ID NO:37.

[0336] Embodiment 280 is the cell or cells of any one of embodiments 274 to 279, wherein the VH comprises a heavy chain framework region (HFR) 1, HFR2, HFR3, and HFR4 and the202666783.1 - 56 -VL comprises a light chain framework region (LFR) 1, LFR2, LFR3, and LFR4, and wherein the HFR1, HFR2, HFR3, and HFR4 comprises an amino acid sequence with at least 80% sequence identity to an HFR1, HFR2, HFR3, and HFR4, respectively, of a VH comprising the amino acid sequence of SEQ ID NO:36, and the LFR1, LFR2, LFR3, and LFR4 comprises an amino acid sequence with at least 80% sequence identity to the LFR1, LFR2, LFR3, and LFR4, respectively, of a VL comprising the amino acid sequence of SEQ ID NO:37.

[0337] Embodiment 281 is the cell or cells of any one of embodiments 274 to 280, wherein the HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4 comprises the amino acid sequence of SEQ ID NOS:208-215, respectively or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:208-215, respectively.

[0338] Embodiment 282 is the cell or cells of embodiment 267, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:45 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:46.

[0339] Embodiment 283 is the cell or cells of embodiment 282, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:45 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:46.

[0340] Embodiment 284 is the cell or cells of embodiment 282 or embodiment 283, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:47-52, respectively.

[0341] Embodiment 285 is the cell or cells of any one of embodiments 282 to 284, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:47-52, respectively.

[0342] Embodiment 286 is the cell or cells of any one of embodiments 282 to 285, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID202666783.1 - 57 -NO:45 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:46.

[0343] Embodiment 287 is the cell or cells of embodiment 286, wherein the VH comprises the amino acid sequence of SEQ ID NO:45 and / or the VL comprises the amino acid sequence of SEQ ID NO:46.

[0344] Embodiment 288 is the cell or cells of any one of embodiments 282 to 287, wherein the VH comprises a heavy chain framework region (HFR) 1, HFR2, HFR3, and HFR4 and the VL comprises a light chain framework region (LFR) 1, LFR2, LFR3, and LFR4, and wherein the HFR1, HFR2, HFR3, and HFR4 comprises an amino acid sequence with at least 80% sequence identity to an HFR1, HFR2, HFR3, and HFR4, respectively, of a VH comprising the amino acid sequence of SEQ ID NO:45, and the LFR1, LFR2, LFR3, and LFR4 comprises an amino acid sequence with at least 80% sequence identity to the LFR1, LFR2, LFR3, and LFR4, respectively, of a VL comprising the amino acid sequence of SEQ ID NO:46.

[0345] Embodiment 289 is the cell or cells of any one of embodiments 282 to 288, wherein the HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4 comprises the amino acid sequence of SEQ ID NOS:216, 209, 217, 211, 218, 219, 220, and 215, respectively or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:216, 209, 217, 211, 218, 219, 220, and 215, respectively.

[0346] Embodiment 290 is the cell or cells of embodiment 267, wherein the antigen binding domain comprises a nanobody, wherein the nanobody comprises a complementarity determining region (CDR) 1, CDR2, and CDR3 having at least 80% sequence identity to the CDR1, CDR2, CDR3 from the nanobody of SEQ ID NO:53.

[0347] Embodiment 291 is the cell or cells of embodiment 290, wherein the nanobody comprises a CDR1, CDR2, and CDR3 having the amino acid sequence of the CDR1, CDR2, and CDR3, respectively, from the nanobody of SEQ ID NO:53.

[0348] Embodiment 292 is the cell or cells of embodiment 290 or embodiment 291, wherein the CDR1, CDR2, and CDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:54-56, respectively.

[0349] Embodiment 293 is the cell or cells of any one of embodiments 290 to 292, wherein the CDR1, CDR2, and CDR3 each comprise the amino acid sequence of SEQ ID NOS:54-56, respectively.

[0350] Embodiment 294 is the cell or cells of any one of embodiments 290 to 293, wherein the nanobody comprises a framework region (FR) 1, FR2, FR3, and FR4, wherein the FR1, FR2, FR3, and FR4 comprises an amino acid sequence with at least 80% sequence identity to202666783.1 - 58 -an FR1, FR2, FR3, and FR4, respectively, of a nanobody comprising the amino acid sequence of SEQ ID NO:53.

[0351] Embodiment 295 is the cell or cells of any one of embodiments 290 to 294, wherein the FR1, FR2, FR3, and FR4 comprises the amino acid sequence of SEQ ID NOS:221-224, respectively or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:221-224, respectively.

[0352] Embodiment 296 is the cell or cells of embodiment 264, wherein the antigen binding domain comprises anti-TYRP-1.

[0353] Embodiment 297 is the cell or cells of embodiment 296, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:62 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:63.

[0354] Embodiment 298 is the cell or cells of embodiment 297, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:62 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:63.

[0355] Embodiment 299 is the cell or cells of embodiment 297 or embodiment 298, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:64-69, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:64-69, respectively.

[0356] Embodiment 300 is the cell or cells of any one of embodiments 297 to 299, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:62 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:63, or wherein the VH comprises the amino acid sequence of SEQ ID NO:62 and / or the VL comprises the amino acid sequence of SEQ ID NO:63.

[0357] Embodiment 301 is the cell or cells of embodiment 264, wherein the antigen binding domain comprises anti-TGF-β.202666783.1 - 59 -

[0358] Embodiment 302 is the cell or cells of embodiment 301, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:71 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:72.

[0359] Embodiment 303 is the cell or cells of embodiment 302, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:71 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:72.

[0360] Embodiment 304 is the cell or cells of embodiment 302 or embodiment 303, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:73-78, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:73-78, respectively.

[0361] Embodiment 305 is the cell or cells of any one of embodiments 302 to 304, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:71 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:72, or wherein the VH comprises the amino acid sequence of SEQ ID NO:71 and / or the VL comprises the amino acid sequence of SEQ ID NO:72.

[0362] Embodiment 306 is the cell or cells of embodiment 301, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:80 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:81.

[0363] Embodiment 307 is the cell or cells of embodiment 306, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and202666783.1 - 60 -HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:80 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:81.

[0364] Embodiment 308 is the cell or cells of embodiment 306 or embodiment 307, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:82-87, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:82-87, respectively.

[0365] Embodiment 309 is the cell or cells of any one of embodiments 306 to 308, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:80 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:81, or wherein the VH comprises the amino acid sequence of SEQ ID NO:80 and / or the VL comprises the amino acid sequence of SEQ ID NO:81.

[0366] Embodiment 310 is the cell or cells of embodiment 301, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:89 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:90.

[0367] Embodiment 311 is the cell or cells of embodiment 310, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:89 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:90.

[0368] Embodiment 312 is the cell or cells of embodiment 310 or embodiment 311, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:91-96, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:91-96, respectively.202666783.1 - 61 -

[0369] Embodiment 313 is the cell or cells of any one of embodiments 310 to 312, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:89 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:90, or wherein the VH comprises the amino acid sequence of SEQ ID NO:89 and / or the VL comprises the amino acid sequence of SEQ ID NO:90.

[0370] Embodiment 314 is the cell or cells of embodiment 264, wherein the antigen binding domain comprises anti-CLDN6.

[0371] Embodiment 315 is the cell or cells of embodiment 314, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:98 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:99.

[0372] Embodiment 316 is the cell or cells of embodiment 315, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:98 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:99.

[0373] Embodiment 317 is the cell or cells of embodiment 315 or embodiment 316, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:100-103, the sequence TSN, and SEQ ID NO:105, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:100- 103, the sequence TSN, and SEQ ID NO:105, respectively.

[0374] Embodiment 318 is the cell or cells of any one of embodiments 315 to 317, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:98 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:99, or wherein the VH comprises the amino acid sequence of SEQ ID NO:98 and / or the VL comprises the amino acid sequence of SEQ ID NO:99.

[0375] Embodiment 319 is the cell or cells of embodiment 264, wherein the antigen binding domain comprises anti-GPC3.202666783.1 - 62 -

[0376] Embodiment 320 is the cell or cells of embodiment 319, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:107 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:108.

[0377] Embodiment 321 is the cell or cells of embodiment 315, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:107 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:108

[0378] Embodiment 322 is the cell or cells of embodiment 315 or embodiment 316, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:109-114, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:109-114, respectively.

[0379] Embodiment 323 is the cell or cells of any one of embodiments 315 to 317, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:107 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:108, or wherein the VH comprises the amino acid sequence of SEQ ID NO:107 and / or the VL comprises the amino acid sequence of SEQ ID NO:108.

[0380] Embodiment 324 is the cell or cells of any one of embodiments 274 to 289 or embodiment 297 to 323, wherein the VH is amino proximal to the VL.

[0381] Embodiment 325 is the cell or cells of any one of embodiments 274 to 289 or embodiment 297 to 323, wherein the VL is amino proximal to the VH.

[0382] Embodiment 326 is the cell or cells of any one of embodiments 274 to 289 or embodiment 297 to 325, wherein the antigen binding domain comprises a linker between the VH and VL.

[0383] Embodiment 327 is the cell or cells of embodiment 326, wherein the linker comprises a glycine serine linker.202666783.1 - 63 -

[0384] Embodiment 328 is the cell or cells of embodiment 327, wherein the linker comprises (G4S)n, wherein n is an integer selected from 1-10.

[0385] Embodiment 329 is the cell or cells of embodiment 328, wherein the linker comprises or consists of SEQ ID NO:119 or 121.

[0386] Embodiment 330 is the cell or cells of embodiment 326, wherein the linker comprises SEQ ID NO:120.

[0387] Embodiment 331 is the cell or cells of any one of embodiments 274 to 330, wherein the antigen binding domain comprises the amino acid sequence of one of SEQ ID NOS:35, 44, 53, 57, 61, 70, 79, 88, 97, or 106 or an amino acid sequence having at least 80% sequence identity to one of SEQ ID NOS:35, 44, 53, 57, 61, 70, 79, 88, 97, or 106.

[0388] Embodiment 332 is the cell or cells of any one of embodiments 229 to 331, further comprising a signal peptide.

[0389] Embodiment 333 is the cell or cells of embodiment 332, wherein the signal peptide comprises SEQ ID NO:122 or 123 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:122 or 123.

[0390] Embodiment 334 is the cell or cells of any one of embodiments 262 to 333, wherein the transmembrane domain is an α or β chain of the T cell receptor or a transmembrane domain from CD28, CD3-epsilon, CD45, CD4, CD5, CD8-α, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD123, CD134, CD137 or CD154.

[0391] Embodiment 335 is the cell or cells of embodiment 334, wherein the transmembrane domain is a CD28 transmembrane domain.

[0392] Embodiment 336 is the cell or cells of any one of embodiments 262 to 335, wherein the transmembrane domain comprises SEQ ID NO:130 or 131 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:130 or 131.

[0393] Embodiment 337 is the cell or cells of any one of embodiments 262 to 336, wherein the intracellular signaling domain comprises a primary signaling domain.

[0394] Embodiment 338 is the cell or cells of embodiment 337, wherein the primary signaling domain is a CD3ζ (zeta) signaling domain.

[0395] Embodiment 339 is the cell or cells of embodiment 337 or embodiment 338, wherein the primary signaling domain comprises SEQ ID NO:134 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:134.

[0396] Embodiment 340 is the cell or cells of any one of embodiments 262 to 339, wherein the intracellular signaling domain further comprises a costimulatory domain.202666783.1 - 64 -

[0397] Embodiment 341 is the cell or cells of embodiment 340, wherein the costimulatory domain comprises a signaling domain from 4-1BB (CD137), CD28, IL-15Rα, OX40, CD2, CD27, CDS, ICAM-1, LFA-1 (CD11a / CD18), or ICOS (CD278).

[0398] Embodiment 342 is the cell or cells of embodiment 341, wherein the costimulatory domain comprises a CD28 or 4-1BB signaling domain.

[0399] Embodiment 343 is the cell or cells of any of embodiments 340 to 342, wherein the costimulatory domain comprises SEQ ID NO:132 or 133 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:132 or 133.

[0400] Embodiment 344 is the cell or cells of any of embodiments 262 to 343, wherein the antigen binding domain and the transmembrane domain are separated by a hinge region.

[0401] Embodiment 345 is the cell or cells of embodiment 344, wherein the hinge region comprises an IgG4 hinge, a CD8α hinge, an IgG1 hinge, or a CD34 hinge.

[0402] Embodiment 346 is the cell or cells of embodiment 344 or embodiment 345, wherein the hinge region is between 8 and 300 amino acids in length.

[0403] Embodiment 347 is the cell or cells of any one of embodiments 344 to 346, wherein the hinge region comprises one of SEQ ID NOS:126-129 or an amino acid sequence having at least 80% sequence identity to one of SEQ ID NOS:126-129.

[0404] Embodiment 348 is the cell or cells of any one of embodiments 237 to 347, wherein the viral vector has integrated into the cell’s genome.

[0405] Embodiment 349 is the cell or cells of any one of embodiments 263 to 348, wherein the CAR is expressed from a nucleic acid comprising a constitutive promoter that directs the expression of the CAR.

[0406] Embodiment 350 is the cell or cells of any one of embodiments 263 to 348, wherein the CAR is expressed from a nucleic acid comprising a conditional promoter that directs the expression of the CAR.

[0407] Embodiment 351 is the cell or cells of embodiment 350, wherein the promoter responds positively to at least one cytokine or to T-cell activation.

[0408] Embodiment 352 is the cell or cells of embodiment 351, where the at least one cytokine is TGF- β, IL-6, TNF α, IFN-γ, IL-1β, IL-2, IL-8, or IL-10 or the promoter responds positively to NFAT or NF-κB.

[0409] Embodiment 353 is the cell or cells of any one of embodiments 227 to 352, wherein the cell(s) comprise a T cell, a natural killer (NK) cell, a natural killer T cell (NKT), an invariant natural killer T cell (iNKT), stem cell, lymphoid progenitor cell, peripheral blood mononuclear202666783.1 - 65 -cell (PBMC), bone marrow cell, fetal liver cell, embryonic stem cell, cord blood cell, induced pluripotent stem cell (iPS cell).

[0410] Embodiment 354 is the cell or cells of embodiment 353, wherein the cell is a T cell or an NK cell.

[0411] Embodiment 355 is the cell or cells of embodiment 354, wherein the T cell comprises a naïve memory T cell.

[0412] Embodiment 356 is the cell or cells of embodiment 355, wherein the naïve memory T cell comprises a CD4+ or CD8+ T cell.

[0413] Embodiment 357 is the cell or cells of any one of embodiments 354 to 356, wherein the T cell comprises a T cell from a population of CD14 depleted, CD25 depleted, and / or CD62L enriched PBMCs.

[0414] Embodiment 358 is the cell or cells of any one of embodiments 227 to 357, wherein the cells are a population of cells comprising 103-108cells.

[0415] Embodiment 359 is a composition comprising the population of cells of embodiment 358, wherein the composition is a pharmaceutically acceptable formulation.

[0416] Embodiment 360 is a method of making a cell comprising introducing into the cell the nucleic acid of any of embodiments 106 to 225.

[0417] Embodiment 361 is the method of embodiment 360, wherein the cell is infected with a virus comprising the nucleic acid.

[0418] Embodiment 362 is the method of embodiment 361, wherein the virus comprises lentivirus or a lentiviral-derived virus or vector.

[0419] Embodiment 363 is the method of any one of embodiments 360 to 362, wherein the cell is a T cell, a natural killer (NK) cell, a natural killer T cell (NKT), an invariant natural killer T cell (iNKT), stem cell, lymphoid progenitor cell, peripheral blood mononuclear cell (PBMC), bone marrow cell, fetal liver cell, embryonic stem cell, cord blood cell, induced pluripotent stem cell (iPS cell).

[0420] Embodiment 364 is the method of embodiment 363, wherein the cell is a T cell or an NK cell.

[0421] Embodiment 365 is the method of embodiment 364, wherein the T cell comprises a naïve memory T cell.

[0422] Embodiment 366 is the method of embodiment 365, wherein the naïve memory T cell comprises a CD4+ or CD8+ T cell.202666783.1 - 66 -

[0423] Embodiment 367 is the method of any one of embodiments 364 to 366, wherein the T cell comprises a T cell from a population of CD14 depleted, CD25 depleted, and / or CD62L enriched PBMCs.

[0424] Embodiment 368 is the method of any one of embodiments 364 to 366, wherein the cell is not yet a T cell or NK cell, the method further comprising culturing the cell under conditions that promote the differentiation of the cell into a T cell or an NK cell.

[0425] Embodiment 369 is the method of any of embodiments 360 to 368, further comprising culturing the cell under conditions to expand the cell before and or after introducing the nucleic acid into the cell.

[0426] Embodiment 370 is the method of embodiment 369, wherein the cell is cultured with serum-free medium.

[0427] Embodiment 371 is a method of treating a patient with cancer comprising administering to the patient an effective amount of the composition of embodiment 359.

[0428] Embodiment 372 is a method for increasing anti-tumor immunity and / or tumor cell cytotoxicity in a patient having cancer, the method comprising administering to the patient an effective amount of the composition of embodiment 359.

[0429] Embodiment 373 is the method of embodiment 371 or embodiment 372, wherein the patient has ovarian, melanoma, glioma, glioblastoma, sarcoma, hepatocellular carcinoma, lung cancer, non-small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, breast cancer, neuroblastoma, thyroid cancer, gastric cancer, ovarian cancer, testis cancer, or uterine cancer.

[0430] Embodiment 374 is the method of any one of embodiments 371 to 373, further comprising administering an additional therapy to the patient.

[0431] Embodiment 375 is the method of embodiment 374, wherein the additional therapy comprises an immunotherapy, anti-angiogenic therapy, chemotherapy, surgery, radiotherapy, neoantigen therapy, or vaccination.

[0432] Embodiment 376 is the method of embodiment 374, wherein the additional therapy comprises immunotherapy and wherein the immunotherapy comprises immune checkpoint inhibitor therapy or bispecific T cell engagers.

[0433] Embodiment 377 is the method of any one of embodiments 371 to 376, wherein the cells are autologous.

[0434] Embodiment 378 is an anti-VEGF antibody or antigen binding fragment comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and202666783.1 - 67 -HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:2 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:3.

[0435] Embodiment 379 is the antibody or antigen binding fragment of embodiment 378, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:2 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:3.

[0436] Embodiment 380 is the antibody or antigen binding fragment of embodiment 378 or embodiment 379, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:4-9, respectively.

[0437] Embodiment 381 is the antibody or antigen binding fragment of any one of embodiments 378 to 380, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:4-9, respectively.

[0438] Embodiment 382 is the antibody or antigen binding fragment of any one of embodiments 378 to 381, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:2 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:3.

[0439] Embodiment 383 is the antibody or antigen binding fragment of embodiment 382, wherein the VH comprises the amino acid sequence of SEQ ID NO:2 and / or the VL comprises the amino acid sequence of SEQ ID NO:3.

[0440] Embodiment 384 is the antibody or antigen binding fragment of any one of embodiments 378 to 383, wherein the antibody or antigen binding fragment comprises a heavy chain framework region (HFR) 1, HFR2, HFR3, and HFR4 and light chain framework region (LFR) 1, LFR2, LFR3, and LFR4, and wherein the HFR1, HFR2, HFR3, and HFR4 comprises an amino acid sequence with at least 80% sequence identity to an HFR1, HFR2, HFR3, and HFR4, respectively, of a VH comprising the amino acid sequence of SEQ ID NO:2, and the LFR1, LFR2, LFR3, and LFR4 comprises an amino acid sequence with at least 80% sequence identity to the LFR1, LFR2, LFR3, and LFR4, respectively, of a VL comprising the amino acid sequence of SEQ ID NO:3.202666783.1 - 68 -

[0441] Embodiment 385 is the antibody or antigen binding fragment of any one of embodiments 378 to 383, wherein the HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4 comprises the amino acid sequence of SEQ ID NOS:10-17, respectively or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:10-17, respectively.

[0442] Embodiment 386 is an anti-VEGF antibody or antigen binding fragment comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:19 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:20.

[0443] Embodiment 387 is the antibody or antigen binding fragment of embodiment 386, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:19 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:20.

[0444] Embodiment 388 is the antibody or antigen binding fragment of embodiment 386 or embodiment 387, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:21-26, respectively.

[0445] Embodiment 389 is the antibody or antigen binding fragment of any one of embodiments 386 to 388, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:21-26, respectively.

[0446] Embodiment 390 is the antibody or antigen binding fragment of any one of embodiments 386 to 389, wherein the VH comprises an amino acid sequence with at least 80% sequence identity to SEQ ID NO:19 and / or the VL comprises an amino acid sequence with at least 80% sequence identity to SEQ ID NO:20.

[0447] Embodiment 391 is the antibody or antigen binding fragment of embodiment 390, wherein the VH comprises the amino acid sequence of SEQ ID NO:19 and / or the VL comprises the amino acid sequence of SEQ ID NO:20.202666783.1 - 69 -

[0448] Embodiment 392 is the antibody or antigen binding fragment of any one of embodiments 386 to 391, wherein the antibody or antigen binding fragment comprises a heavy chain framework region (HFR) 1, HFR2, HFR3, and HFR4 and light chain framework region (LFR) 1, LFR2, LFR3, and LFR4, and wherein the HFR1, HFR2, HFR3, and HFR4 comprises an amino acid sequence with at least 80% sequence identity to an HFR1, HFR2, HFR3, and HFR4, respectively, of a VH comprising the amino acid sequence of SEQ ID NO:19, and the LFR1, LFR2, LFR3, and LFR4 comprises an amino acid sequence with at least 80% sequence identity to the LFR1, LFR2, LFR3, and LFR4, respectively, of a VL comprising the amino acid sequence of SEQ ID NO:20.

[0449] Embodiment 393 is the antibody or antigen binding fragment of any one of embodiments 386 to 391, wherein the HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4 comprises the amino acid sequence of SEQ ID NOS:27-34, respectively or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:27-34, respectively.

[0450] Embodiment 394 is the antibody or antigen binding fragment of any one of embodiments 386 to 393, wherein the antibody comprises a VH having at least 70% sequence identity to SEQ ID NO:19 and / or a VL having at least 70% sequence identity to SEQ ID NO:20.

[0451] Embodiment 395 is the antibody or antigen binding fragment of embodiment 394, wherein the antibody comprises a VH of SEQ ID NO:19 and / or a VL of SEQ ID NO:20.

[0452] Embodiment 396 is the antibody of any one of embodiments 378 to 395, wherein the antibody is human, chimeric, or humanized.

[0453] Embodiment 397 is the antibody or antigen-binding fragment of any one of embodiments 378 to 396, wherein the antibody or antigen binding fragment is antagonistic.

[0454] Embodiment 398 is the antibody or antigen binding fragment of any one of embodiments 378 to 397, wherein the antibody is a human antibody, humanized antibody, recombinant antibody, chimeric antibody, an antibody derivative, a veneered antibody, a diabody, a monoclonal antibody, a single domain antibody, or a single chain antibody.

[0455] Embodiment 399 is the antigen binding fragment of any one of embodiments 378 to 397, wherein the antigen binding fragment is a single chain variable fragment (scFv), F(ab’)2, Fab’, Fab, Fv, or rIgG.

[0456] Embodiment 400 is the antigen binding fragment of embodiment 399, wherein the antigen binding fragment comprises a scFv.

[0457] Embodiment 401 is the antigen binding fragment of embodiment 399 or embodiment 400, wherein the VH is amino proximal to the VL.202666783.1 - 70 -

[0458] Embodiment 402 is the antigen binding fragment of embodiment 399 or embodiment 400, wherein the VL is amino proximal to the VH.

[0459] Embodiment 403 is the antigen binding fragment of any one of embodiments 399 to 402, wherein the antigen binding fragment comprises a VH and a VL and a linker between the VH and VL.

[0460] Embodiment 404 is the antigen binding fragment of embodiment 403, wherein the linker comprises a glycine serine linker.

[0461] Embodiment 405 is the antigen binding fragment of embodiment 404, wherein the linker comprises (G4S)n, wherein n is an integer selected from 1-10.

[0462] Embodiment 406 is the antigen binding fragment of embodiment 405, wherein the linker comprises or consists of SEQ ID NO:119 or 121.

[0463] Embodiment 407 is the antigen binding fragment of embodiment 403, wherein the linker comprises SEQ ID NO:120.

[0464] Embodiment 408 is the antibody or antigen binding fragment of any one of embodiments 378 to 407, wherein the VL comprises an arginine at the carboxy terminus of the VL.

[0465] Embodiment 409 is the antigen binding fragment of any one of embodiments 400 to 408, wherein the antigen binding fragment comprises the amino acid sequence of SEQ ID NO:1 or 18 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:1 or 18.

[0466] Embodiment 410 is a polypeptide comprising the antigen binding fragment of any one of embodiments 378 to 409.

[0467] Embodiment 411 is the polypeptide of embodiment 410, wherein the polypeptide comprises at least two antigen binding fragments, wherein each antigen binding fragment is independently selected from an antigen binding fragment of any one of embodiments 378 to 409.

[0468] Embodiment 412 is the polypeptide of embodiment 410 or embodiment 411, wherein the polypeptide is multivalent.

[0469] Embodiment 413 is the polypeptide of any one of embodiments 410 to 412, wherein the polypeptide is bispecific.

[0470] Embodiment 414 is a chimeric polypeptide comprising: (a) an antigen binding fragment of any one of embodiments 378 to 409; (b) a transmembrane domain; and (c) an intracellular signaling domain.202666783.1 - 71 -

[0471] Embodiment 415 is the chimeric polypeptide of embodiment 414, wherein the antigen binding fragment comprises SEQ ID NO:1 or SEQ ID NO:18 or an amino acid sequence having at least 70% sequence identity to SEQ ID NO:1 or SEQ ID NO:18.

[0472] Embodiment 416 is the chimeric polypeptide of embodiment 414 or embodiment 415, further comprising a signal peptide.

[0473] Embodiment 417 is the chimeric polypeptide of embodiment 416, wherein the signal peptide comprises SEQ ID NO:122 or 123 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:122 or 123.

[0474] Embodiment 418 is the chimeric polypeptide of any one of embodiments 414 to 417, wherein the transmembrane domain is an α or β chain of the T cell receptor or a transmembrane domain from CD28, CD3-epsilon, CD45, CD4, CD5, CD8-α, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD123, CD134, CD137 or CD154.

[0475] Embodiment 419 is the chimeric polypeptide of embodiment 418, wherein the transmembrane domain is a CD28 transmembrane domain.

[0476] Embodiment 420 is the chimeric polypeptide of any one of embodiments 414 to 419, wherein the transmembrane domain comprises SEQ ID NO:130 or 131 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:130 or 131.

[0477] Embodiment 421 is the chimeric polypeptide of any one of embodiments 414 to 420, wherein the intracellular signaling domain comprises a primary signaling domain.

[0478] Embodiment 422 is the chimeric polypeptide of embodiment 421, wherein the primary signaling domain is a CD3ζ (zeta) signaling domain.

[0479] Embodiment 423 is the chimeric polypeptide of embodiment 421 or embodiment 422, wherein the primary signaling domain comprises SEQ ID NO:134 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:134.

[0480] Embodiment 424 is the chimeric polypeptide of any one of embodiments 414 to 423, wherein the intracellular signaling domain further comprises a costimulatory domain.

[0481] Embodiment 425 is the chimeric polypeptide of embodiment 424, wherein the costimulatory domain comprises a signaling domain from 4-1BB (CD137), CD28, IL-15Rα, OX40, CD2, CD27, CDS, ICAM-1, LFA-1 (CD11a / CD18), or ICOS (CD278).

[0482] Embodiment 426 is the chimeric polypeptide of embodiment 425, wherein the costimulatory domain comprises a CD28 or 4-1BB signaling domain.

[0483] Embodiment 427 is the chimeric polypeptide of any of embodiments 424 to 426, wherein the costimulatory domain comprises SEQ ID NO:132 or 133 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:132 or 133.202666783.1 - 72 -

[0484] Embodiment 428 is the chimeric polypeptide of any of embodiments 414 to 427, wherein the antigen binding domain and the transmembrane domain are separated by a hinge region.

[0485] Embodiment 429 is the chimeric polypeptide of embodiment 428, wherein the hinge region comprises an IgG4 hinge, a CD8α hinge, an IgG1 hinge, or a CD34 hinge.

[0486] Embodiment 430 is the chimeric polypeptide of embodiment 428 or embodiment 429, wherein the hinge region is between 8 and 300 amino acids in length.

[0487] Embodiment 431 is the chimeric polypeptide of any one of embodiments 428 to 430, wherein the hinge region comprises one of SEQ ID NOS:126-129 or an amino acid sequence having at least 80% sequence identity to one of SEQ ID NOS:126-129.

[0488] Embodiment 432 is a fusion protein comprising the antigen binding fragment of any one of embodiments 378 to 409 and a CD3 binding region.

[0489] Embodiment 433 is the fusion protein of embodiment 432, wherein the CD3 binding region comprises a CD3-specific fragment antigen binding (Fab), single chain variable fragment (scFv), single domain antibody, or single chain antibody.

[0490] Embodiment 434 is the antibody or antigen binding fragment of any one of embodiments 378 to 409, polypeptide of any one of embodiments 410 to 431, or the fusion protein of embodiment 432 or embodiment 433, wherein the antibody, antigen binding fragment, polypeptide, or fusion protein is conjugated to a detection or therapeutic agent.

[0491] Embodiment 435 is the antibody, antigen binding fragment, polypeptide, or fusion protein of embodiment 434, wherein the agent comprises a fluorescent molecule, radiative molecule, or toxin.

[0492] Embodiment 436 is one or more nucleic acids encoding the antibody or antigen binding fragment of any one of embodiments 378 to 409, the polypeptide of any one of embodiments 410 to 431, or the fusion protein of any one of embodiments 432 to 435.

[0493] Embodiment 437 is the nucleic acid(s) of embodiment 436, wherein the nucleic acid is an expression construct.

[0494] Embodiment 438 is the nucleic acid(s) of embodiment 437, wherein the expression construct is a plasmid or viral vector.

[0495] Embodiment 439 is the nucleic acid(s) of embodiment 438, wherein the viral vector is a vector derived from a retrovirus or a vector derived from a lentivirus.

[0496] Embodiment 440. The nucleic acid(s) of any one of embodiments 436 to 439, wherein the nucleic acid comprises one or more promoter(s) that direct the expression of the antibody, antigen binding fragment, or polypeptide.202666783.1 - 73 -

[0497] Embodiment 441 is the nucleic acid(s) of embodiment 440, wherein the promoter is constitutive.

[0498] Embodiment 442 is the nucleic acid(s) of embodiment 440, wherein the promoter is conditional.

[0499] Embodiment 443 is the nucleic acid(s) of embodiment 442, wherein the promoter responds positively to at least one cytokine or to T-cell activation.

[0500] Embodiment 444 is the nucleic acid(s) of embodiment 443, where the at least one cytokine is TGF- β, IL-6, TNF α, IFN-γ, IL-1β, IL-2, IL-8, or IL-10 or the promoter responds positively to NFAT or NF-κB.

[0501] Embodiment 445 is the nucleic acid of any one of embodiments 378 to 444, wherein the nucleic acid encodes for a chimeric antigen receptor (CAR), wherein the CAR comprises: (a) an antigen binding domain; (b) a transmembrane domain; and (c) an intracellular signaling domain.

[0502] Embodiment 446 is the nucleic acid of embodiment 445, wherein the antigen binding domain comprises an anti-TYRP-1, anti-IL-13Rα2, anti-TGF-β, anti-CLDN6, anti- GPC3, or anti-MSLN antigen binding domain.

[0503] Embodiment 447 is the nucleic acid of embodiment 446, wherein the antigen binding domain comprises an anti-IL-13Rα2 comprising an IL-13 mutein polypeptide.

[0504] Embodiment 448 is the nucleic acid of embodiment 446, wherein the IL-13 mutein polypeptide comprises the amino acid sequence of SEQ ID NO:115 or 233, or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:115 or 233.

[0505] Embodiment 449 is the nucleic acid of embodiment 446, wherein the antigen binding domain comprises anti-MSLN.

[0506] Embodiment 450 is the nucleic acid of embodiment 449, wherein the antigen binding domain comprises a nanobody, wherein the nanobody comprises a complementarity determining region (CDR) 1, CDR2, and CDR3 having at least 80% sequence identity to the CDR1, CDR2, CDR3 from the nanobody of SEQ ID NO:57.

[0507] Embodiment 451 is the nucleic acid of embodiment 450, wherein the nanobody comprises a CDR1, CDR2, and CDR3 having the amino acid sequence of the CDR1, CDR2, and CDR3, respectively, from the nanobody of SEQ ID NO:57.

[0508] Embodiment 452 is the nucleic acid of embodiment 450 or embodiment 451, wherein the CDR1, CDR2, and CDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:58-60, respectively.202666783.1 - 74 -

[0509] Embodiment 453 is the nucleic acid of any one of embodiments 450 to 452, wherein the CDR1, CDR2, and CDR3 each comprise the amino acid sequence of SEQ ID NOS:58-60, respectively.

[0510] Embodiment 454 is the nucleic acid of any one of embodiments 450 to 453, wherein the nanobody comprises a framework region (FR) 1, FR2, FR3, and FR4, wherein the FR1, FR2, FR3, and FR4 comprises an amino acid sequence with at least 80% sequence identity to an FR1, FR2, FR3, and FR4, respectively, of a nanobody comprising the amino acid sequence of SEQ ID NO:57.

[0511] Embodiment 455 is the nucleic acid of embodiment 454, wherein the FR1, FR2, FR3, and FR4 comprises the amino acid sequence of SEQ ID NOS:225-228, respectively or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:225-228, respectively.

[0512] Embodiment 456 is the nucleic acid of embodiment 449, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:36 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:37.

[0513] Embodiment 457 is the nucleic acid of embodiment 456, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:36 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:37.

[0514] Embodiment 458 is the nucleic acid of embodiment 456 or embodiment 457, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:38-43, respectively.

[0515] Embodiment 459 is the nucleic acid of any one of embodiments 456 to 458, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:38-43, respectively.202666783.1 - 75 -

[0516] Embodiment 460 is the nucleic acid of any one of embodiments 456 to 459, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:36 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:37.

[0517] Embodiment 461 is the nucleic acid of embodiment 460, wherein the VH comprises the amino acid sequence of SEQ ID NO:36 and / or the VL comprises the amino acid sequence of SEQ ID NO:37.

[0518] Embodiment 462 is the nucleic acid of any one of embodiments 456 to 461, wherein the VH comprises a heavy chain framework region (HFR) 1, HFR2, HFR3, and HFR4 and the VL comprises a light chain framework region (LFR) 1, LFR2, LFR3, and LFR4, and wherein the HFR1, HFR2, HFR3, and HFR4 comprises an amino acid sequence with at least 80% sequence identity to an HFR1, HFR2, HFR3, and HFR4, respectively, of a VH comprising the amino acid sequence of SEQ ID NO:36, and the LFR1, LFR2, LFR3, and LFR4 comprises an amino acid sequence with at least 80% sequence identity to the LFR1, LFR2, LFR3, and LFR4, respectively, of a VL comprising the amino acid sequence of SEQ ID NO:37.

[0519] Embodiment 463 is the nucleic acid of any one of embodiments 456 to 462, wherein the HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4 comprises the amino acid sequence of SEQ ID NOS:208-215, respectively or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:208-215, respectively.

[0520] Embodiment 464 is the nucleic acid of embodiment 449, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:45 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:46.

[0521] Embodiment 465 is the nucleic acid of embodiment 464, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:45 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:46.202666783.1 - 76 -

[0522] Embodiment 466 is the nucleic acid of embodiment 464 or embodiment 465, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:47-52, respectively.

[0523] Embodiment 467 is the nucleic acid of any one of embodiments 464 to 466, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:47-52, respectively.

[0524] Embodiment 468 is the nucleic acid of any one of embodiments 464 to 467, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:45 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:46.

[0525] Embodiment 469 is the nucleic acid of embodiment 468, wherein the VH comprises the amino acid sequence of SEQ ID NO:45 and / or the VL comprises the amino acid sequence of SEQ ID NO:46.

[0526] Embodiment 470 is the nucleic acid of any one of embodiments 464 to 469, wherein the VH comprises a heavy chain framework region (HFR) 1, HFR2, HFR3, and HFR4 and the VL comprises a light chain framework region (LFR) 1, LFR2, LFR3, and LFR4, and wherein the HFR1, HFR2, HFR3, and HFR4 comprises an amino acid sequence with at least 80% sequence identity to an HFR1, HFR2, HFR3, and HFR4, respectively, of a VH comprising the amino acid sequence of SEQ ID NO:45, and the LFR1, LFR2, LFR3, and LFR4 comprises an amino acid sequence with at least 80% sequence identity to the LFR1, LFR2, LFR3, and LFR4, respectively, of a VL comprising the amino acid sequence of SEQ ID NO:46.

[0527] Embodiment 471 is the nucleic acid of any one of embodiments 464 to 470, wherein the HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4 comprises the amino acid sequence of SEQ ID NOS:216, 209, 217, 211, 218, 219, 220, and 215, respectively or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:216, 209, 217, 211, 218, 219, 220, and 215, respectively.

[0528] Embodiment 472 is the nucleic acid of embodiment 449, wherein the antigen binding domain comprises a nanobody, wherein the nanobody comprises a complementarity determining region (CDR) 1, CDR2, and CDR3 having at least 80% sequence identity to the CDR1, CDR2, CDR3 from the nanobody of SEQ ID NO:53.

[0529] Embodiment 473 is the nucleic acid of embodiment 472, wherein the nanobody comprises a CDR1, CDR2, and CDR3 having the amino acid sequence of the CDR1, CDR2, and CDR3, respectively, from the nanobody of SEQ ID NO:53.202666783.1 - 77 -

[0530] Embodiment 474 is the nucleic acid of embodiment 472 or embodiment 473, wherein the CDR1, CDR2, and CDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:54-56, respectively.

[0531] Embodiment 475 is the nucleic acid of any one of embodiments 472 to 474, wherein the CDR1, CDR2, HCDR2, LCDR1, LCDR2, and CDR3 each comprise the amino acid sequence of SEQ ID NOS:54-56, respectively.

[0532] Embodiment 476 is the nucleic acid of any one of embodiments 472 to 475, wherein the nanobody comprises a framework region (FR) 1, FR2, FR3, and FR4, wherein the FR1, FR2, FR3, and FR4 comprises an amino acid sequence with at least 80% sequence identity to an FR1, FR2, FR3, and FR4, respectively, of a nanobody comprising the amino acid sequence of SEQ ID NO:53.

[0533] Embodiment 477 is the nucleic acid of any one of embodiments 472 to 476, wherein the FR1, FR2, FR3, and FR4 comprises the amino acid sequence of SEQ ID NOS:221-224, respectively or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:221-224, respectively.

[0534] Embodiment 478 is the nucleic acid of embodiment 446, wherein the antigen binding domain comprises anti-TYRP-1.

[0535] Embodiment 479 is the nucleic acid of embodiment 478, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:62 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:63.

[0536] Embodiment 480 is the nucleic acid of embodiment 479, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:62 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:63.

[0537] Embodiment 481 is the nucleic acid of embodiment 479 or embodiment 480, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:64-69,202666783.1 - 78 -respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:64-69, respectively.

[0538] Embodiment 482 is the nucleic acid of any one of embodiments 479 to 481, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:62 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:63, or wherein the VH comprises the amino acid sequence of SEQ ID NO:62 and / or the VL comprises the amino acid sequence of SEQ ID NO:63.

[0539] Embodiment 483 is the nucleic acid of embodiment 446, wherein the antigen binding domain comprises anti-TGF-β.

[0540] Embodiment 484 is the nucleic acid of embodiment 483, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:71 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:72.

[0541] Embodiment 485 is the nucleic acid of embodiment 484, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:71 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:72.

[0542] Embodiment 486 is the nucleic acid of embodiment 484 or embodiment 485, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:73-78, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:73-78, respectively.

[0543] Embodiment 487 is the nucleic acid of any one of embodiments 484 to 486, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:71 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:72, or wherein the VH comprises the amino acid sequence of SEQ ID NO:71 and / or the VL comprises the amino acid sequence of SEQ ID NO:72.202666783.1 - 79 -

[0544] Embodiment 488 is the nucleic acid of embodiment 483, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:80 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:81.

[0545] Embodiment 489 is the nucleic acid of embodiment 488, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:80 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:81.

[0546] Embodiment 490 is the nucleic acid of embodiment 488 or embodiment 489, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:82-87, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:82-87, respectively.

[0547] Embodiment 491 is the nucleic acid of any one of embodiments 488 to 490, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:80 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:81, or wherein the VH comprises the amino acid sequence of SEQ ID NO:80 and / or the VL comprises the amino acid sequence of SEQ ID NO:81.

[0548] Embodiment 492 is the nucleic acid of embodiment 483, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:89 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:90.

[0549] Embodiment 493 is the nucleic acid of embodiment 492, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and202666783.1 - 80 -HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:89 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:90.

[0550] Embodiment 494 is the nucleic acid of embodiment 492 or embodiment 493, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:91-96, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:91-96, respectively.

[0551] Embodiment 495 is the nucleic acid of any one of embodiments 492 to 494, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:89 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:90, or wherein the VH comprises the amino acid sequence of SEQ ID NO:89 and / or the VL comprises the amino acid sequence of SEQ ID NO:90.

[0552] Embodiment 496 is the nucleic acid of embodiment 446, wherein the antigen binding domain comprises anti-CLDN6.

[0553] Embodiment 497 is the nucleic acid of embodiment 496, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:98 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:99.

[0554] Embodiment 498 is the nucleic acid of embodiment 497, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:98 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:99.

[0555] Embodiment 499 is the nucleic acid of embodiment 497 or embodiment 498, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:100-103, the sequence TSN, and SEQ ID NO:105, respectively or wherein the HCDR1, HCDR2, HCDR3,202666783.1 - 81 -LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:100- 103, the sequence TSN, and SEQ ID NO:105, respectively.

[0556] Embodiment 500 is the nucleic acid of any one of embodiments 497 to 499, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:98 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:99, or wherein the VH comprises the amino acid sequence of SEQ ID NO:98 and / or the VL comprises the amino acid sequence of SEQ ID NO:99.

[0557] Embodiment 501 is the nucleic acid of embodiment 446, wherein the antigen binding domain comprises anti-GPC3.

[0558] Embodiment 502 is the nucleic acid of embodiment 501, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:107 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:108.

[0559] Embodiment 503 is the nucleic acid of embodiment 497, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:107 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:108

[0560] Embodiment 504 is the nucleic acid of embodiment 497 or embodiment 498, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:109-114, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:109-114, respectively.

[0561] Embodiment 505 is the nucleic acid of any one of embodiments 497 to 499, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:107 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:108, or wherein the VH comprises the amino acid sequence of SEQ ID NO:107 and / or the VL comprises the amino acid sequence of SEQ ID NO:108.202666783.1 - 82 -

[0562] Embodiment 506 is the nucleic acid of any one of embodiments 456 to 471 or embodiments 479 to 505, wherein the VH is amino proximal to the VL.

[0563] Embodiment 507 is the nucleic acid of any one of embodiments 456 to 471 or embodiments 479 to 505, wherein the VL is amino proximal to the VH.

[0564] Embodiment 508 is the nucleic acid of any one of embodiments 456 to 471 or embodiments 479 to 507, wherein the antigen binding domain comprises a linker between the VH and VL.

[0565] Embodiment 509 is the nucleic acid of embodiment 508, wherein the linker comprises a glycine serine linker.

[0566] Embodiment 510 is the nucleic acid of embodiment 509, wherein the linker comprises (G4S)n, wherein n is an integer selected from 1-10.

[0567] Embodiment 511 is the nucleic acid of embodiment 510, wherein the linker comprises or consists of SEQ ID NO:119 or 121.

[0568] Embodiment 512 is the nucleic acid of embodiment 508, wherein the linker comprises SEQ ID NO:120.

[0569] Embodiment 513 is the nucleic acid of any one of embodiments 456 to 512, wherein the antigen binding domain comprises the amino acid sequence of one of SEQ ID NOS:35, 44, 53, 57, 61, 70, 79, 88, 97, or 106 or an amino acid sequence having at least 80% sequence identity to one of SEQ ID NOS:35, 44, 53, 57, 61, 70, 79, 88, 97, or 106.

[0570] Embodiment 514 is the nucleic acid of any one of embodiments 378 to 513, wherein the nucleic acid is DNA or is RNA.

[0571] Embodiment 515 is the nucleic acid of any one of embodiments 378 to 514, further comprising a signal peptide.

[0572] Embodiment 516 is the nucleic acid of embodiment 515, wherein the signal peptide comprises SEQ ID NO:122 or 123 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:122 or 123.

[0573] Embodiment 517 is the nucleic acid of any one of embodiments 445 to 516, wherein the transmembrane domain is an α or β chain of the T cell receptor or a transmembrane domain from CD28, CD3-epsilon, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD123, CD134, CD137 or CD154.

[0574] Embodiment 518 is the nucleic acid of embodiment 517, wherein the transmembrane domain is a CD28 transmembrane domain.202666783.1 - 83 -

[0575] Embodiment 519 is the nucleic acid of any one of embodiments 445 to 518, wherein the transmembrane domain comprises SEQ ID NO:130 or 131 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:130 or 131.

[0576] Embodiment 520 is the nucleic acid of any one of embodiments 445 to 519, wherein the intracellular signaling domain comprises a primary signaling domain.

[0577] Embodiment 521 is the nucleic acid of embodiment 520, wherein the primary signaling domain is a CD3ζ (zeta) signaling domain.

[0578] Embodiment 522 is the nucleic acid of embodiment 520 or embodiment 521, wherein the primary signaling domain comprises SEQ ID NO:134 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:134.

[0579] Embodiment 523 is the nucleic acid of any one of embodiments 445 to 522, wherein the intracellular signaling domain further comprises a costimulatory domain.

[0580] Embodiment 524 is the nucleic acid of embodiment 523, wherein the costimulatory domain comprises a signaling domain from 4-1BB (CD137), CD28, IL-15Rα, OX40, CD2, CD27, CDS, ICAM-1, LFA-1 (CD11a / CD18), or ICOS (CD278).

[0581] Embodiment 525 is the nucleic acid of embodiment 524, wherein the costimulatory domain comprises a CD28 or 4-1BB signaling domain.

[0582] Embodiment 526 is the nucleic acid of any of embodiments 523 to 525, wherein the costimulatory domain comprises SEQ ID NO:132 or 133 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:132 or 133.

[0583] Embodiment 527 is the nucleic acid of any of embodiments 445 to 526, wherein the antigen binding domain and the transmembrane domain are separated by a hinge region.

[0584] Embodiment 528 is the nucleic acid of embodiment 527, wherein the hinge region comprises an IgG4 hinge, a CD8α hinge, an IgG1 hinge, or a CD34 hinge.

[0585] Embodiment 529 is the nucleic acid of embodiment 527 or embodiment 528, wherein the hinge region is between 8 and 300 amino acids in length.

[0586] Embodiment 530 is the nucleic acid of any one of embodiments 527 to 529, wherein the hinge region comprises one of SEQ ID NOS:126-129 or an amino acid sequence having at least 80% sequence identity to one of SEQ ID NOS:126-129.

[0587] Embodiment 531 is the nucleic acid of any one of embodiments 436 to 530, wherein the nucleic acid further encodes an IL-12 polypeptide and an IL-18 polypeptide.

[0588] Embodiment 532 is the nucleic acid of any one of embodiments 436 to 530, wherein the nucleic acid further encodes an IL-12, IFN-β, and / or CXCL9 polypeptide.202666783.1 - 84 -

[0589] Embodiment 533 is the nucleic acid of embodiment 531 or embodiment 532, wherein the IL-12 polypeptide comprises a p40 and / or p35 subunit.

[0590] Embodiment 534 is the nucleic acid of any one of embodiments 531 to 533, wherein the IL-12 polypeptide comprises a single-chain IL-12 polypeptide.

[0591] Embodiment 535 is the nucleic acid of embodiment 534, wherein the p40 subunit is amino-proximal to the p35 subunit.

[0592] Embodiment 536 is the nucleic acid of embodiment 534, wherein the p35 subunit is amino-proximal to the p40 subunit.

[0593] Embodiment 537 is the nucleic acid of any one of embodiments 436 to 536, wherein the nucleic acid further encodes for a suicide gene product.

[0594] Embodiment 538 is the nucleic acid of embodiment 537, wherein suicide gene product is herpes simplex virus thymidine kinase (HSV-TK), purine nucleoside phosphorylase (PNP), cytosine deaminase (CD), carboxypetidase G2, cytochrome P450, linamarase, β- lactamase, nitroreductase (NTR), carboxypeptidase A, split Cas9, truncated EGFR, or inducible caspase 9.

[0595] Embodiment 539 is the nucleic acid of any one of embodiments 531 to 538, wherein the nucleic acid encodes for a linker between the p40 and p35 subunit.

[0596] Embodiment 540 is the nucleic acid of any one of embodiments 445 to 539, wherein the nucleic acid comprises one or more promoter(s) that direct the expression of the suicide gene product, CAR, IL-12 polypeptide, IFN-β polypeptide, CXCL9 polypeptide, and / or IL-18 polypeptide.

[0597] Embodiment 541 is the nucleic acid of embodiment 540, wherein the promoter is constitutive.

[0598] Embodiment 542 is the nucleic acid of embodiment 540, wherein the promoter is conditional.

[0599] Embodiment 543 is the nucleic acid of embodiment 542, wherein the promoter responds positively to at least one cytokine or to T-cell activation.

[0600] Embodiment 544 is the nucleic acid of embodiment 543, where the at least one cytokine is TGF- β, IL-6, TNF α, IFN-γ, IL-1β, IL-2, IL-8, or IL-10 or the promoter responds positively to NFAT or NF-κB.

[0601] Embodiment 545 is the nucleic acid of any one of embodiments 531 to 544, wherein the IL-12, IFN-β, CXCL9, and / or IL-18 polypeptide is derived from a human IL-12, human IFN-β, human CXCL9, and / or human IL-18 polypeptide.202666783.1 - 85 -

[0602] Embodiment 546 is the nucleic acid of any one of embodiments 531 to 545, wherein the IL-18 polypeptide comprises a decoy resistant IL-18 polypeptide.

[0603] Embodiment 547 is the nucleic acid of any one of embodiments 533 to 546, wherein the p40 polypeptide comprises the amino acid sequence of SEQ ID NO:236 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:236.

[0604] Embodiment 548 is the nucleic acid of any one of embodiments 533 to 547, wherein the p35 polypeptide comprises the amino acid sequence of SEQ ID NO:237 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:237.

[0605] Embodiment 549 is the nucleic acid of any one of embodiments 531 to 548, wherein the IL-12 polypeptide comprises the amino acid sequence of SEQ ID NO:230 or 234 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:230 or 234.

[0606] Embodiment 550 is the nucleic acid of any one of embodiments 531 to 549, wherein the IL-18 polypeptide comprises the amino acid sequence of SEQ ID NO:229, 232, or 238-240 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:229, 232, or 238-240.

[0607] Embodiment 551 is the nucleic acid of any one of embodiments 531 to 550, wherein the IFN-β polypeptide comprises the amino acid sequence of SEQ ID NO:253 or 255 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:253 or 255.

[0608] Embodiment 552 is the nucleic acid of any one of embodiments 531 to 551, wherein the CXCL9 polypeptide comprises the amino acid sequence of SEQ ID NO:257 or 259 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:257 or 259.

[0609] Embodiment 553 is the nucleic acid of any one of embodiments 531 to 552, wherein the nucleic acid encodes for a signal sequence at the amino-terminus of the IL-12, IFN-β, CXCL9 and / or IL-18 polypeptide.

[0610] Embodiment 554 is the nucleic acid of embodiment 553, wherein the signal sequence comprises SEQ ID NO:122, 123, 235, 241, 254, 256, 258, and / or 260.

[0611] Embodiment 555 is the nucleic acid of any one of embodiments 531 to 554, wherein the IL-12 polypeptide and the IL-18 polypeptide are expressed from one nucleic acid.

[0612] Embodiment 556 is the nucleic acid of embodiment 555, wherein the nucleic acid expresses a polypeptide comprising the IL-12 polypeptide and the IL-18 polypeptide.

[0613] Embodiment 557 is the nucleic acid of any one of embodiments 531 to 554, wherein the IL-12 polypeptide, IFN-β polypeptide, and the CXCL9 polypeptide are expressed from one nucleic acid.202666783.1 - 86 -

[0614] Embodiment 558 is the nucleic acid of embodiment 557, wherein the nucleic acid expresses a polypeptide comprising the IL-12, IFN-β, and CXCL9 polypeptide.

[0615] Embodiment 559 is the nucleic acid of embodiment 389 or embodiment 390, wherein the nucleic acid encodes for a self-cleaving peptide or internal ribosome entry sequence (IRES), wherein the self-cleaving peptide or IRES is between the IL-12, IL-18, CXCL9, and / or IFN-β polypeptides.

[0616] Embodiment 560 is the nucleic acid of embodiment 559, wherein the self-cleaving peptide comprises SEQ ID NO:135 or 136.

[0617] Embodiment 561 is a polypeptide expressed from and / or encoded by the nucleic acid of any one of embodiments 436 to 560.

[0618] Embodiment 562 is a cell or population of cells comprising the nucleic acid of any one of embodiments 436 to 560 or the polypeptide of embodiment 561.

[0619] Embodiment 563 is the cell or cells of embodiment 562, wherein the cell or cells express a polypeptide encoded by the nucleic acid.

[0620] Embodiment 564 is the cell or cells of embodiment 562 or embodiment 563, wherein the cell further comprises a heterologous nucleic acid encoding an IL-12 polypeptide and a heterologous nucleic acid encoding an IL-18 polypeptide.

[0621] Embodiment 565 is the cell or cells of embodiment 562 or embodiment 563, wherein the cell further comprises a heterologous nucleic acid encoding an IL-12 polypeptide, a heterologous nucleic acid encoding an IFN-β polypeptide and a heterologous nucleic acid encoding a CXCL9 polypeptide.

[0622] Embodiment 566 is the cell of embodiment 564 or embodiment 565, wherein the IL-12 polypeptide comprises a p40 and / or p35 subunit.

[0623] Embodiment 567 is the cell of any one of embodiments 564 to 566, wherein the IL- 12 polypeptide comprises a single-chain IL-12 polypeptide.

[0624] Embodiment 568 is the cell of embodiment 567, wherein the p40 subunit is amino- proximal to the p35 subunit.

[0625] Embodiment 569 is the cell of embodiment 567, wherein the p35 subunit is amino- proximal to the p40 subunit.

[0626] Embodiment 570 is the cell of any one of embodiments 567 to 569, wherein the IL- 12 polypeptide comprises a linker between the p40 and p35 subunit.

[0627] Embodiment 571 is the cell or cells of any one of embodiments 564 to 570, wherein the nucleic acid encoding the IL-12 polypeptide, IL-18 polypeptide, IFN-β polypeptide, CXCL9 polypeptide, and / or anti-VEGF antigen binding fragment is an expression construct.202666783.1 - 87 -

[0628] Embodiment 572 is the cell or cells of embodiment 571, wherein the expression construct is a plasmid or viral vector.

[0629] Embodiment 573 is the cell or cells of embodiment 572, wherein the viral vector is a vector derived from a retrovirus or a vector derived from a lentivirus.

[0630] Embodiment 574 is the cell or cells of any one of embodiments 562 to 573, wherein the cell comprises a nucleic acid encoding for a suicide gene product.

[0631] Embodiment 575 is the cell or cells of embodiment 574, wherein suicide gene product is herpes simplex virus thymidine kinase (HSV-TK), purine nucleoside phosphorylase (PNP), cytosine deaminase (CD), carboxypetidase G2, cytochrome P450, linamarase, β- lactamase, nitroreductase (NTR), carboxypeptidase A, split Cas9, or inducible caspase 9.

[0632] Embodiment 576 is the cell or cells of any one of embodiments 562 to 575, wherein the nucleic acid comprises one or more promoter(s) that direct the expression of the suicide gene product, CAR, polypeptide, IL-12 polypeptide, IL-18 polypeptide, IFN-β polypeptide, CXCL9 polypeptide, and / or anti-VEGF antigen binding fragment.

[0633] Embodiment 577 is the cell or cells of embodiment 576, wherein the promoter is constitutive.

[0634] Embodiment 578 is the cell or cells of embodiment 576, wherein the promoter is conditional.

[0635] Embodiment 579 is the cell or cells of embodiment 578, wherein the promoter responds positively to at least one cytokine or to T-cell activation.

[0636] Embodiment 580 is the cell or cells of embodiment 579, where the at least one cytokine is TGF- β, IL-6, TNF α, IFN-γ, IL-1β, IL-2, IL-8, or IL-10 or the promoter responds positively to NFAT or NF-κB.

[0637] Embodiment 581 is the cell or cells of any one of embodiments 564 to 580, wherein the IL-12, IFN- β, CXCL9, and / or IL-18 polypeptide is derived from a human IL-12, human IFN- β, human CXCL9, and / or human IL-18 polypeptide.

[0638] Embodiment 582 is the cell or cells of any one of embodiments 564 to 581, wherein the IL-18 polypeptide comprises a decoy resistant IL-18 polypeptide.

[0639] Embodiment 583 is the cell or cells of any one of embodiments 566 to 582, wherein the p40 polypeptide comprises the amino acid sequence of SEQ ID NO:236 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:236.

[0640] Embodiment 584 is the cell or cells of any one of embodiments 566 to 583, wherein the p35 polypeptide comprises the amino acid sequence of SEQ ID NO:237 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:237.202666783.1 - 88 -

[0641] Embodiment 585 is the cell or cells of any one of embodiments 564 to 584, wherein the IL-12 polypeptide comprises the amino acid sequence of SEQ ID NO:230 or 234 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:230 or 234.

[0642] Embodiment 586 is the cell or cells of any one of embodiments 564 to 585, wherein the IL-18 polypeptide comprises the amino acid sequence of SEQ ID NO:229, 232, or 238-240 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:229, 232, or 238-240.

[0643] Embodiment 587 is the cell or cells of any one of embodiments 564 to 586, wherein the IFN-β polypeptide comprises the amino acid sequence of SEQ ID NO:253 or 255 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:253 or 255.

[0644] Embodiment 588. cell or cells of any one of embodiments 564 to 587, wherein the CXCL9 polypeptide comprises the amino acid sequence of SEQ ID NO:257 or 259 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:257 or 259.

[0645] Embodiment 589 is the cell or cells of any one of embodiments 564 to 586, wherein the nucleic acid encodes for a signal sequence at the amino-terminus of the IL-12, IFN-β, CXCL9 and / or IL-18 polypeptide.

[0646] Embodiment 590 is the cell or cells of any one of embodiments 564 to 589, wherein the signal sequence comprises SEQ ID NO:122, 123, 235, 241, 254, 256, 258, and / or 260.

[0647] Embodiment 591 is the cell or cells of any one of embodiments 564 to 590, wherein the IL-12 polypeptide, IL-18 polypeptide, and / or anti-VEGF antigen binding fragment are expressed from one nucleic acid.

[0648] Embodiment 592 is the cell or cells of embodiment 591, wherein the nucleic acid expresses a polypeptide comprising the IL-12 polypeptide, IL-18 polypeptide, and / or anti- VEGF antigen binding fragment.

[0649] Embodiment 593 is the cell or cells of any one of embodiments 564 to 590, wherein the IL-12 polypeptide, IFN-β polypeptide, CXCL9 polypeptide, and / or anti-VEGF antigen binding fragment are expressed from one nucleic acid.

[0650] Embodiment 594 is the nucleic acid of embodiment 593, wherein the nucleic acid expresses a polypeptide comprising the IL-12, IFN-β, CXCL9, , and / or anti-VEGF antigen binding fragment.

[0651] Embodiment 595 is the cell or cells of any one of embodiments 591 to 594, wherein the nucleic acid encodes for a self-cleaving peptide or internal ribosome entry sequence (IRES), wherein the self-cleaving peptide or IRES is between the IL-12 polypeptide, IL-18 polypeptide, IFN-β, polypeptide, CXCL9 polypeptide, and / or anti-VEGF antigen binding fragment.202666783.1 - 89 -

[0652] Embodiment 596 is the cell or cells of embodiment 595, wherein the self-cleaving polypeptide comprises SEQ ID NO:135 or 136.

[0653] Embodiment 597 is the cell or cells of any one of embodiments 564 to 596, wherein the cell further comprises a nucleic acid encoding a CAR or TCR.

[0654] Embodiment 598 is the cell or cells of embodiment 597, wherein the nucleic acid encodes for a chimeric antigen receptor (CAR), wherein the CAR comprises: (a) an antigen binding domain; (b) a transmembrane domain; and (c) an intracellular signaling domain.

[0655] Embodiment 599 is the cell or cells embodiment 598, wherein the antigen binding domain comprises an anti-TYRP-1, anti-IL-13Rα2, anti-TGF-β, anti-CLDN6, anti-GPC3, or anti-MSLN antigen binding domain.

[0656] Embodiment 600 is the cell or cells of embodiment 599, wherein the antigen binding domain comprises an anti-IL-13Rα2 comprising an IL-13 mutein polypeptide.

[0657] Embodiment 601 is the cell or cells of embodiment 599, wherein the IL-13 mutein polypeptide comprises the amino acid sequence of SEQ ID NO:115 or 233, or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:115 or 233.

[0658] Embodiment 602 is the cell or cells of embodiment 599, wherein the antigen binding domain comprises anti-MSLN.

[0659] Embodiment 603 is the cell or cells of embodiment 602, wherein the antigen binding domain comprises a nanobody, wherein the nanobody comprises a complementarity determining region (CDR) 1, CDR2, and CDR3 having at least 80% sequence identity to the CDR1, CDR2, CDR3 from the nanobody of SEQ ID NO:57.

[0660] Embodiment 604 is the cell or cells of embodiment 603, wherein the nanobody comprises a CDR1, CDR2, and CDR3 having the amino acid sequence of the CDR1, CDR2, and CDR3, respectively, from the nanobody of SEQ ID NO:57.

[0661] Embodiment 605 is the cell or cells of embodiment 603 or embodiment 604, wherein the CDR1, CDR2, and CDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:58-60, respectively.

[0662] Embodiment 606 is the cell or cells of any one of embodiments 603 to 605, wherein the CDR1, CDR2, and CDR3 each comprise the amino acid sequence of SEQ ID NOS:58-60, respectively.

[0663] Embodiment 607 is the cell or cells of any one of embodiments 603 to 606, wherein the nanobody comprises a framework region (FR) 1, FR2, FR3, and FR4, wherein the FR1, FR2, FR3, and FR4 comprises an amino acid sequence with at least 80% sequence identity to202666783.1 - 90 -an FR1, FR2, FR3, and FR4, respectively, of a nanobody comprising the amino acid sequence of SEQ ID NO:57.

[0664] Embodiment 608 is the cell or cells of embodiment 607, wherein the FR1, FR2, FR3, and FR4 comprises the amino acid sequence of SEQ ID NOS:225-228, respectively or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:225-228, respectively.

[0665] Embodiment 609 is the cell or cells of embodiment 602, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:36 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:37.

[0666] Embodiment 610 is the cell or cells of embodiment 609, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:36 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:37.

[0667] Embodiment 611 is the cell or cells of embodiment 609 or embodiment 610, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:38-43, respectively.

[0668] Embodiment 612 is the cell or cells of any one of embodiments 609 to 611, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:38-43, respectively.

[0669] Embodiment 613 is the cell or cells of any one of embodiments 609 to 612, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:36 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:37.

[0670] Embodiment 614 is the cell or cells of embodiment 613, wherein the VH comprises the amino acid sequence of SEQ ID NO:36 and / or the VL comprises the amino acid sequence of SEQ ID NO:37.202666783.1 - 91 -

[0671] Embodiment 615 is the cell or cells of any one of embodiments 609 to 614, wherein the VH comprises a heavy chain framework region (HFR) 1, HFR2, HFR3, and HFR4 and the VL comprises a light chain framework region (LFR) 1, LFR2, LFR3, and LFR4, and wherein the HFR1, HFR2, HFR3, and HFR4 comprises an amino acid sequence with at least 80% sequence identity to an HFR1, HFR2, HFR3, and HFR4, respectively, of a VH comprising the amino acid sequence of SEQ ID NO:36, and the LFR1, LFR2, LFR3, and LFR4 comprises an amino acid sequence with at least 80% sequence identity to the LFR1, LFR2, LFR3, and LFR4, respectively, of a VL comprising the amino acid sequence of SEQ ID NO:37.

[0672] Embodiment 616 is the cell or cells of any one of embodiments 609 to 615, wherein the HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4 comprises the amino acid sequence of SEQ ID NOS:208-215, respectively or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:208-215, respectively.

[0673] Embodiment 617 is the cell or cells of embodiment 602, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:45 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:46.

[0674] Embodiment 618 is the cell or cells of embodiment 617, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:45 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:46.

[0675] Embodiment 619 is the cell or cells of embodiment 617 or embodiment 618, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:47-52, respectively.

[0676] Embodiment 620 is the cell or cells of any one of embodiments 617 to 619, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:47-52, respectively.202666783.1 - 92 -

[0677] Embodiment 621 is the cell or cells of any one of embodiments 617 to 620, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:45 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:46.

[0678] Embodiment 622 is the cell or cells of embodiment 621, wherein the VH comprises the amino acid sequence of SEQ ID NO:45 and / or the VL comprises the amino acid sequence of SEQ ID NO:46.

[0679] Embodiment 623 is the cell or cells of any one of embodiments 617 to 622, wherein the VH comprises a heavy chain framework region (HFR) 1, HFR2, HFR3, and HFR4 and the VL comprises a light chain framework region (LFR) 1, LFR2, LFR3, and LFR4, and wherein the HFR1, HFR2, HFR3, and HFR4 comprises an amino acid sequence with at least 80% sequence identity to an HFR1, HFR2, HFR3, and HFR4, respectively, of a VH comprising the amino acid sequence of SEQ ID NO:45, and the LFR1, LFR2, LFR3, and LFR4 comprises an amino acid sequence with at least 80% sequence identity to the LFR1, LFR2, LFR3, and LFR4, respectively, of a VL comprising the amino acid sequence of SEQ ID NO:46.

[0680] Embodiment 624 is the cell or cells of any one of embodiments 617 to 623, wherein the HFR1, HFR2, HFR3, HFR4, LFR1, LFR2, LFR3, and LFR4 comprises the amino acid sequence of SEQ ID NOS:216, 209, 217, 211, 218, 219, 220, and 215, respectively or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:216, 209, 217, 211, 218, 219, 220, and 215, respectively.

[0681] Embodiment 625 is the cell or cells of embodiment 602, wherein the antigen binding domain comprises a nanobody, wherein the nanobody comprises a complementarity determining region (CDR) 1, CDR2, and CDR3 having at least 80% sequence identity to the CDR1, CDR2, CDR3 from the nanobody of SEQ ID NO:53.

[0682] Embodiment 626 is the cell or cells of embodiment 625, wherein the nanobody comprises a CDR1, CDR2, and CDR3 having the amino acid sequence of the CDR1, CDR2, and CDR3, respectively, from the nanobody of SEQ ID NO:53.

[0683] Embodiment 627 is the cell or cells of embodiment 625 or embodiment 626, wherein the CDR1, CDR2, and CDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:54-56, respectively.

[0684] Embodiment 628 is the cell or cells of any one of embodiments 625 to 627, wherein the CDR1, CDR2, and CDR3 each comprise the amino acid sequence of SEQ ID NOS:54-56, respectively.202666783.1 - 93 -

[0685] Embodiment 629 is the cell or cells of any one of embodiments 625 to 628, wherein the nanobody comprises a framework region (FR) 1, FR2, FR3, and FR4, wherein the FR1, FR2, FR3, and FR4 comprises an amino acid sequence with at least 80% sequence identity to an FR1, FR2, FR3, and FR4, respectively, of a nanobody comprising the amino acid sequence of SEQ ID NO:53.

[0686] Embodiment 630 is the cell or cells of any one of embodiments 625 to 629, wherein the FR1, FR2, FR3, and FR4 comprises the amino acid sequence of SEQ ID NOS:221-224, respectively or an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:221-224, respectively.

[0687] Embodiment 631 is the cell or cells of embodiment 599, wherein the antigen binding domain comprises anti-TYRP-1.

[0688] Embodiment 632 is the cell or cells of embodiment 631, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:62 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:63.

[0689] Embodiment 633 is the cell or cells of embodiment 632, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:62 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:63.

[0690] Embodiment 634 is the cell or cells of embodiment 632 or embodiment 633, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:64-69, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:64-69, respectively.

[0691] Embodiment 635 is the cell or cells of any one of embodiments 632 to 634, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:62 and / or the VL comprises an amino acid sequence with at least 70% sequence identity202666783.1 - 94 -to SEQ ID NO:63, or wherein the VH comprises the amino acid sequence of SEQ ID NO:62 and / or the VL comprises the amino acid sequence of SEQ ID NO:63.

[0692] Embodiment 636 is the cell or cells of embodiment 599, wherein the antigen binding domain comprises anti-TGF-β.

[0693] Embodiment 637 is the cell or cells of embodiment 636, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:71 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:72.

[0694] Embodiment 638 is the cell or cells of embodiment 637, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:71 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:72.

[0695] Embodiment 639 is the cell or cells of embodiment 637 or embodiment 638, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:73-78, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:73-78, respectively.

[0696] Embodiment 640 is the cell or cells of any one of embodiments 637 to 639, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:71 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:72, or wherein the VH comprises the amino acid sequence of SEQ ID NO:71 and / or the VL comprises the amino acid sequence of SEQ ID NO:72.

[0697] Embodiment 641 is the cell or cells of embodiment 636, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:80 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1,202666783.1 - 95 -LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:81.

[0698] Embodiment 642 is the cell or cells of embodiment 641, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:80 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:81.

[0699] Embodiment 643 is the cell or cells of embodiment 641 or embodiment 642, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:82-87, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:82-87, respectively.

[0700] Embodiment 644 is the cell or cells of any one of embodiments 641 to 643, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:80 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:81, or wherein the VH comprises the amino acid sequence of SEQ ID NO:80 and / or the VL comprises the amino acid sequence of SEQ ID NO:81.

[0701] Embodiment 645 is the cell or cells of embodiment 636, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:89 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:90.

[0702] Embodiment 646 is the cell or cells of embodiment 645, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:89 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:90.

[0703] Embodiment 647 is the cell or cells of embodiment 645 or embodiment 646, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an202666783.1 - 96 -amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:91-96, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:91-96, respectively.

[0704] Embodiment 648 is the cell or cells of any one of embodiments 645 to 647, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:89 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:90, or wherein the VH comprises the amino acid sequence of SEQ ID NO:89 and / or the VL comprises the amino acid sequence of SEQ ID NO:90.

[0705] Embodiment 649 is the cell or cells of embodiment 599, wherein the antigen binding domain comprises anti-CLDN6.

[0706] Embodiment 650 is the cell or cells of embodiment 649, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:98 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:99.

[0707] Embodiment 651 is the cell or cells of embodiment 650, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:98 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:99.

[0708] Embodiment 652 is the cell or cells of embodiment 650 or embodiment 651, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:100-103, the sequence TSN, and SEQ ID NO:105, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:100- 103, the sequence TSN, and SEQ ID NO:105, respectively.

[0709] Embodiment 653 is the cell or cells of any one of embodiments 650 to 652, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:98 and / or the VL comprises an amino acid sequence with at least 70% sequence identity202666783.1 - 97 -to SEQ ID NO:99, or wherein the VH comprises the amino acid sequence of SEQ ID NO:98 and / or the VL comprises the amino acid sequence of SEQ ID NO:99.

[0710] Embodiment 654 is the cell or cells of embodiment 599, wherein the antigen binding domain comprises anti-GPC3.

[0711] Embodiment 655 is the cell or cells of embodiment 654, wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:107 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:108.

[0712] Embodiment 656 is the cell or cells of embodiment 650, wherein the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:107 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:108

[0713] Embodiment 657 is the cell or cells of embodiment 650 or embodiment 651, wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:109-114, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:109-114, respectively.

[0714] Embodiment 658 is the cell or cells of any one of embodiments 650 to 652, wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:107 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:108, or wherein the VH comprises the amino acid sequence of SEQ ID NO:107 and / or the VL comprises the amino acid sequence of SEQ ID NO:108.

[0715] Embodiment 659 is the cell or cells of any one of embodiments 609 to 624 or embodiments 632 to 658, wherein the VH is amino proximal to the VL.

[0716] Embodiment 660 is the cell or cells of any one of embodiments 609 to 624 or embodiments 632 to 658, wherein the VL is amino proximal to the VH.202666783.1 - 98 -

[0717] Embodiment 661 is the cell or cells of any one of embodiments 609 to 624 or embodiments 632 to 660, wherein the antigen binding domain comprises a linker between the VH and VL.

[0718] Embodiment 662 is the cell or cells of embodiment 661, wherein the linker comprises a glycine serine linker.

[0719] Embodiment 663 is the cell or cells of embodiment 662, wherein the linker comprises (G4S)n, wherein n is an integer selected from 1-10.

[0720] Embodiment 664 is the cell or cells of embodiment 663, wherein the linker comprises or consists of SEQ ID NO:119 or 121.

[0721] Embodiment 665 is the cell or cells of embodiment 661, wherein the linker comprises SEQ ID NO:120.

[0722] Embodiment 666 is the cell or cells of any one of embodiments 609 to 665, wherein the antigen binding domain comprises the amino acid sequence of one of SEQ ID NOS:35, 44, 53, 57, 61, 70, 79, 88, 97, or 106 or an amino acid sequence having at least 80% sequence identity to one of SEQ ID NOS:35, 44, 53, 57, 61, 70, 79, 88, 97, or 106.

[0723] Embodiment 667 is the cell or cells of any one of embodiments 564 to 666, further comprising a signal peptide.

[0724] Embodiment 668 is the cell or cells of embodiment 667, wherein the signal peptide comprises SEQ ID NO:122 or 123 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:122 or 123.

[0725] Embodiment 669 is the cell or cells of any one of embodiments 597 to 668, wherein the transmembrane domain is an α or β chain of the T cell receptor or a transmembrane domain from CD28, CD3-epsilon, CD45, CD4, CD5, CD8-α, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD123, CD134, CD137 or CD154.

[0726] Embodiment 670 is the cell or cells of embodiment 669, wherein the transmembrane domain is a CD28 transmembrane domain.

[0727] Embodiment 671 is the cell or cells of any one of embodiments 597 to 670, wherein the transmembrane domain comprises SEQ ID NO:130 or 131 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:130 or 131.

[0728] Embodiment 672 is the cell or cells of any one of embodiments 597 to 671, wherein the intracellular signaling domain comprises a primary signaling domain.

[0729] Embodiment 673 is the cell or cells of embodiment 672, wherein the primary signaling domain is a CD3ζ (zeta) signaling domain.202666783.1 - 99 -

[0730] Embodiment 674 is the cell or cells of embodiment 672 or embodiment 673, wherein the primary signaling domain comprises SEQ ID NO:134 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:134.

[0731] Embodiment 675 is the cell or cells of any one of embodiments 597 to 674, wherein the intracellular signaling domain further comprises a costimulatory domain.

[0732] Embodiment 676 is the cell or cells of embodiment 675, wherein the costimulatory domain comprises a signaling domain from 4-1BB (CD137), CD28, IL-15Rα, OX40, CD2, CD27, CDS, ICAM-1, LFA-1 (CD11a / CD18), or ICOS (CD278).

[0733] Embodiment 677 is the cell or cells of embodiment 676, wherein the costimulatory domain comprises a CD28 or 4-1BB signaling domain.

[0734] Embodiment 678 is the cell or cells of any of embodiments 675 to 677, wherein the costimulatory domain comprises SEQ ID NO:132 or 133 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:132 or 133.

[0735] Embodiment 679 is the cell or cells of any of embodiments 597 to 678, wherein the antigen binding domain and the transmembrane domain are separated by a hinge region.

[0736] Embodiment 680 is the cell or cells of embodiment 679, wherein the hinge region comprises an IgG4 hinge, a CD8α hinge, an IgG1 hinge, or a CD34 hinge.

[0737] Embodiment 681 is the cell or cells of embodiment 679 or embodiment 680, wherein the hinge region is between 8 and 300 amino acids in length.

[0738] Embodiment 682 is the cell or cells of any one of embodiments 679 to 681, wherein the hinge region comprises one of SEQ ID NOS:126-129 or an amino acid sequence having at least 80% sequence identity to one of SEQ ID NOS:126-129.

[0739] Embodiment 683 is the cell or cells of any one of embodiments 572 to 683, wherein the viral vector has integrated into the cell’s genome.

[0740] Embodiment 684 is the cell or cells of any one of embodiments 562 to 683, wherein the cell(s) comprise a T cell, a natural killer (NK) cell, a natural killer T cell (NKT), an invariant natural killer T cell (iNKT), stem cell, lymphoid progenitor cell, peripheral blood mononuclear cell (PBMC), bone marrow cell, fetal liver cell, embryonic stem cell, cord blood cell, induced pluripotent stem cell (iPS cell).

[0741] Embodiment 685 is the cell or cells of embodiment 684, wherein the cell is a T cell or an NK cell.

[0742] Embodiment 686 is the cell or cells of embodiment 685, wherein the T cell comprises a naïve memory T cell.202666783.1 - 100 -

[0743] Embodiment 687 is the cell or cells of embodiment 686, wherein the naïve memory T cell comprises a CD4+ or CD8+ T cell.

[0744] Embodiment 688 is the cell or cells of any one of embodiments 685 to 687, wherein the T cell comprises a T cell from a population of CD14 depleted, CD25 depleted, and / or CD62L enriched PBMCs.

[0745] Embodiment 689 is the cell or cells of any one of embodiments 562 to 688, wherein the cells are a population of cells comprising 103-1012cells.

[0746] Embodiment 690 is a composition comprising the cells of any one of embodiments 562 to 688 or population of cells of embodiment 689, wherein the composition is a pharmaceutically acceptable formulation.

[0747] Embodiment 691 is a method of making a cell comprising introducing into a cell the nucleic acid of any of embodiments 436 to 560.

[0748] Embodiment 692 is the method of embodiment 691, wherein the cell is infected with a virus comprising the nucleic acid.

[0749] Embodiment 693 is the method of embodiment 692, wherein the virus comprises lentivirus or a lentiviral-derived virus or vector.

[0750] Embodiment 694 is the method of any one of embodiments 691 to 693, wherein the cell is a T cell, a natural killer (NK) cell, a natural killer T cell (NKT), an invariant natural killer T cell (iNKT), stem cell, lymphoid progenitor cell, peripheral blood mononuclear cell (PBMC), bone marrow cell, fetal liver cell, embryonic stem cell, cord blood cell, induced pluripotent stem cell (iPS cell).

[0751] Embodiment 695 is the method of embodiment 694, wherein the cell is a T cell or an NK cell.

[0752] Embodiment 696 is the method of embodiment 695, wherein the T cell comprises a naïve memory T cell.

[0753] Embodiment 697 is the method of embodiment 696, wherein the naïve memory T cell comprises a CD4+ or CD8+ T cell.

[0754] Embodiment 698 is the method of any one of embodiments 695 to 697, wherein the T cell comprises a T cell from a population of CD14 depleted, CD25 depleted, and / or CD62L enriched PBMCs.

[0755] Embodiment 699 is the method of any one of embodiments 695 to 697, wherein the cell is not yet a T cell or NK cell, the method further comprising culturing the cell under conditions that promote the differentiation of the cell into a T cell or an NK cell.202666783.1 - 101 -

[0756] Embodiment 700 is the method of any of embodiments 691 to 699, further comprising culturing the cell under conditions to expand the cell before and or after introducing the nucleic acid into the cell.

[0757] Embodiment 701 is the method of embodiment 700, wherein the cell is cultured with serum-free medium.

[0758] Embodiment 702 is a method of treating a patient with cancer comprising administering to the patient an effective amount of one or more compositions according to embodiments 516, 359, and 690.

[0759] Embodiment 703 is a method for increasing anti-tumor immunity and / or tumor cell cytotoxicity in a patient having cancer, the method comprising administering to the patient an effective amount of one or more compositions according to embodiments 516, 359, and 690.

[0760] Embodiment 704 is the method of embodiment 702 or embodiment 703, wherein the patient has ovarian, melanoma, glioma, glioblastoma, sarcoma, hepatocellular carcinoma, lung cancer, non-small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, breast cancer, neuroblastoma, thyroid cancer, gastric cancer, ovarian cancer, testis cancer, or uterine cancer.

[0761] Embodiment 705 is the method of any one of embodiments 702 to 704, further comprising administering an additional therapy to the patient.

[0762] Embodiment 706 is the method of embodiment 705, wherein the additional therapy comprises an immunotherapy, anti-angiogenic therapy, chemotherapy, surgery, radiotherapy, neoantigen therapy, or vaccination.

[0763] Embodiment 707 is the method of embodiment 705, wherein the additional therapy comprises immunotherapy and wherein the immunotherapy comprises immune checkpoint inhibitor therapy or bispecific T cell engagers.

[0764] Embodiment 708 is the method of any one of embodiments 702 to 707, wherein the cells are autologous.

[0765] Throughout this application, the term “about” is used to indicate that a value includes the inherent variation of error for the measurement or quantitation method.

[0766] The use of the word “a” or “an” when used in conjunction with the term “comprising” may mean “one,” but it is also consistent with the meaning of “one or more,” “at least one,” and “one or more than one.”

[0767] The phrase “and / or” means “and” or “or”. To illustrate, A, B, and / or C includes: A alone, B alone, C alone, a combination of A and B, a combination of A and C, a combination202666783.1 - 102 -of B and C, or a combination of A, B, and C. In other words, “and / or” operates as an inclusive or.

[0768] The words “comprising” (and any form of comprising, such as “comprise” and “comprises”), “having” (and any form of having, such as “have” and “has”), “including” (and any form of including, such as “includes” and “include”) or “containing” (and any form of containing, such as “contains” and “contain”) are inclusive or open-ended and do not exclude additional, unrecited elements or method steps.

[0769] The compositions and methods for their use can “comprise,” “consist essentially of,” or “consist of” any of the ingredients or steps disclosed throughout the specification. Compositions and methods “consisting essentially of” any of the ingredients or steps disclosed limits the scope of the claim to the specified materials or steps which do not materially affect the basic and novel characteristic of the claimed invention. As used in this specification and claim(s), the words “comprising” (and any form of comprising, such as “comprise” and “comprises”), “having” (and any form of having, such as “have” and “has”), “including” (and any form of including, such as “includes” and “include”) or “containing” (and any form of containing, such as “contains” and “contain”) are inclusive or open-ended and do not exclude additional, unrecited elements or method steps. It is contemplated that embodiments described herein in the context of the term “comprising” may also be implemented in the context of the term “consisting of” or “consisting essentially of.”

[0770] It is specifically contemplated that any limitation discussed with respect to one embodiment of the invention may apply to any other embodiment of the invention. Furthermore, any composition of the invention may be used in any method of the invention, and any method of the invention may be used to produce or to utilize any composition of the invention. Aspects of an embodiment set forth in the Examples are also embodiments that may be implemented in the context of embodiments discussed elsewhere in a different Example or elsewhere in the application, such as in the Summary of Invention, Detailed Description of the Embodiments, Claims, and description of Figure Legends.

[0771] Other objects, features, and advantages of the present invention will become apparent from the following detailed description. It should be understood, however, that the detailed description and the specific examples, while indicating specific embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.202666783.1 - 103 -BRIEF DESCRIPTION OF THE DRAWINGS

[0772] The following drawings form part of the present specification and are included to further demonstrate certain aspects of the present invention. The invention may be better understood by reference to one or more of these drawings in combination with the detailed description of specific embodiments presented herein.

[0773] FIG.1A-1B. A panel of MSLN CARs (constructs 1-4 in invention report) utilizing either scFvs or nanobodies were expressed in primary human T cells via retroviral transduction. (A) MSLN CAR expression on retrovirally transduced primary human T cells. Representative flow cytometry plots showing FLAG (CAR) and EGFRt expression. (B) Comparison of MSLN CAR expression on retrovirally transduced primary human T cells. Transduction efficiency (EGFRt expression) and CAR (FLAG) expression comparison among panel of MSLN CARs. The CAR utilizing the NbC6 nanobody was the most well-expressed based on both percentage of retrovirally-transduced T cells expressing the CAR and median fluorescent intensity (MFI) of CAR+ T cells. Thus, the CAR utilizing this NbC6 Nb, hereby referred to as just the MSLN CAR, was selected for subsequent studies. Data are mean ± standard deviation with n = 3 technical replicates. RV**** in (B) corresponds to the labels shown in (A).

[0774] FIG.2A-2C. The MSLN CAR can recognize both murine and human forms of the MSLN antigen. (A) Human T cells expressing the MSLN CAR (MSLN CAR-huT cells) divide in response to murine MSLN. CellTrace Violet (CTV)-dye–stained MSLN CAR-huT cells were co-cultured with MSLN+ ID8 cells (mouse ovarian carcinoma cells). Reduction in CTV fluorescent intensity, indicative of T-cell division, was only observed when MSLN CAR-T cells were co-cultured with ID8 cells. CTV dilution was not observed when MSLN CAR-T cells were cultured in the absence of target cells or when mock-transduced T cells were co- cultured with ID8 target cells, demonstrating cell division by MSLN CAR-T cells in the presence of ID8 is an antigen-specific response. (B) MSLN CAR-huT cells are activated in response to murine MSLN. T-cell activation markers (CD25 and CD137) were upregulated in MSLN CAR-huT cells after a 24-hour co-culture with murine ID8 cells, indicating antigen- specific activation of the CAR-T cells. Effector-to-target (E:T) ratios of 4:1 and 8:1 were tested. Percent positivity (top) and fold-change in median fluorescence intensity (MFI) of the activation markers compared to MFI prior to co-culture (bottom) are shown. (C) MSLN CAR- huT cells are activated in response to human MSLN. T-cell activation markers (CD25 and202666783.1 - 104 -CD137) were upregulated in MSLN CAR-huT cells after a 26.5-hour co-culture with MSLN+ Panc 04.03 (human pancreatic adenocarcinoma) tumor cells indicating antigen-specific activation of the CAR-T cells. E:T ratios of 4:1 and 8:1 were tested. Percent positivity (top) and fold-change in MFI of the activation markers compared to MFI prior to co-culture is shown. Data are mean ± 1 standard deviation with n = 3 (B, C).

[0775] FIG. 3. MSLN CAR-huT cells lyse both human and murine MSLN+ tumor cells. The MSLN CAR enables antigen-specific lysis of MSLN+ human and murine tumor cells in vitro. MSLN CAR-huT cells were co-cultured with murine ID8 tumor cells, human SKOV-3 (human ovarian adenocarcinoma) tumor cells, and human Panc 04.03 tumor cells for 94-, 94-, and 70-hours, respectively. E:T ratios of 4:1 and 8:1 were tested. % lysis is calculated by normalizing the CAR-T cell cytotoxicity to that of the mock-transduced T cells. Data are mean ± 1 standard deviation with n = 3.

[0776] FIG.4A-4C. The MSLN CAR can be expressed and is functional in murine T cells (MSLN CAR-mT cells). (A) MSLN CAR can be expressed in murine T cells. Representative flow cytometry plot demonstrating FLAG (MSLN CAR) expression in bulk CD3+ T cells isolated from the spleen of a C57BL / 6J mouse via retroviral transduction. (B) MSLN CAR- mT cells are activated in response to MSLN+ tumor cells. T-cell activation markers (CD25 and CD137) were upregulated in MSLN CAR-mT cells when co-cultured with ID8 tumor cells at an E:T ratio of 8:1. The fold-change in MFI of the activation markers prior to co-culture is shown. (C) MSLN CAR-mT cells lyse MSLN+ tumor cells. MSLN CAR-mT cells were co- cultured with ID8 tumor cells for 96 hours and % lysis was calculated. E:T ratios of 4:1 and 8:1 were tested. % lysis is calculated by normalizing the CAR-T cell cytotoxicity to that of the mock-transduced T cells. Data are mean ± 1 standard deviation with n = 3 (B, C).

[0777] FIG.5A-5C. MSLN CAR-mT cells show anti-tumor activity in vivo. (A) Schematic of in vivo study.5e6 CAR+ MSLN CAR-mT cells were administered intraperitoneally (i.p.) 6 days after i.p. injection with a mixed dose of 8.5e6 ID8 and 0.5e6 ID8VEGF tumor cells. ID8VEGF tumor cells are ID8 ovarian cancer cells that have been retrovirally transduced to overexpress vascular endothelial growth factor (VEGF), which increases the aggressiveness of the tumor. The tumor cells express firefly luciferase, and tumor burden was monitored via luciferin injection and bioluminescent imaging (BLI) using the IVIS system. (B) MSLN CAR- mT cells lyse tumor cells in vivo. MSLN CAR-mT cells can substantially delay tumor progression in vivo as measured by BLI. Untransduced (UTD) T cells were used as the negative control. (C) MSLN CAR-mT cells significantly improve overall mouse survival compared to202666783.1 - 105 -the UTD control treatment. n = 10 for UTD and n = 15 for MSLN CAR-mT cell treatment. Log-rank test was used to determine significance.

[0778] FIG.6. MSLN CAR-huT cells are capable of lysing patient-derived primary ovarian cancer cells in vitro. MSLN CAR-huT cells were co-cultured with primary ovarian cancer cells harvested from a patient with recurrent and platinum-resistant ovarian cancer for 70 hours at E:T ratios ranging from 1:8 to 8:1. % lysis is calculated by normalizing the CAR-T cell cytotoxicity to that of the non-antigen targeting control CAR-T cells. Data are mean ± standard deviation with n = 3.

[0779] FIG.7. Armored CAR-T cell supernatants reprogram murine macrophages in vitro. To evaluate the ability of the CAR-T cell produced immunomodulatory proteins to positively influence murine macrophages in vitro, murine T cells were transduced with a panel of constructs encoding the IL-13Rα2 CAR and different immunomodulatory proteins. After in vitro culturing, under conditions that stimulate the inducible promoter, supernatants were collected. In parallel, in vitro differentiated murine bone marrow derived macrophages were polarized to an immunosuppressive phenotype with the cytokines IL-4 and IL-13, and tumor- conditioned media (supernatant taken from culturing GL261 cells, a mouse glioblastoma multiform cell line). Subsequently, the immunosuppressive macrophages were cultured in the supernatant collected from the CAR-T cells, such that they would be influenced by factors secreted by the CAR-T cells. After 48 hrs, the phenotype of the macrophages was assessed for upregulation of markers associated with a pro-inflammatory response. As a negative control, “null” refers to a CAR construct that does not encode for any secreted immunomodulatory protein. Averages of triplicates are shown, with error bars representing ± 1 standard deviation.

[0780] FIG.8A-8B. Armored CAR-T cell supernatants enhance murine NK cell activation and tumor cell lysis in vitro. The ability of the immunomodulatory proteins to positively impact primary murine Natural Killer (NK) cell activity in vitro was assessed. Murine T cells were transduced with a panel of constructs encoding the TYRP1 CAR and different immunomodulatory proteins. After in vitro culturing, under conditions that stimulate the inducible promoter, supernatants were collected. In parallel, primary murine NK cells were isolated by negative selection from C57BL / 6J spleens. Subsequently, the NK cells were co- cultured with B16F10 (murine melanoma cell line) cells in the supernatants collected from the CAR-T cells. After 48 hours, (A) activation of the natural killer cells (by CD69 and 4-1BB expression) and (B) killing of the tumor cells were assessed. Averages of triplicates are shown, with error bars representing ± 1 standard deviation.202666783.1 - 106 -

[0781] FIG.9. IL-12 / DR-18 / anti–IL-10 armored TYRP-1.28z CAR-T cells are efficacious against orthotopic syngeneic melanoma tumors in vivo. Ten pooled combinations of TYPR-1 CAR-T cells were pitted against syngeneic B16F10 melanoma tumors orthotopically in vivo. C57BL / 6J mice were implanted with 1e5 B16F10 melanoma cells subcutaneously. After 6 days, mice were given 2e6 pooled CAR+ T cells intravenously (IV). To generate the treatment, primary murine T cells were transduced with constructs that encode the TYPR-1 CAR and 1 immunomodulatory protein. After ex vivo expansion, and prior to IV injection, these CAR-T cells were mixed together at a 1:1:1 ratio, forming triple combinations. Tumor progression was tracked via caliper measurements. Mice were euthanized when tumors exceeded 1500 mm3. As a negative control, “scFv-less” CAR-T cells were injected, wherein the targeting domain of the TYRP-1 CAR has been removed from the null-output construct. Kaplan-Meier survival curve is shown. Statistics are calculated by log-rank analysis, with Bonferroni correction for multiple comparisons.

[0782] FIG. 10A–10B. IL-12 / DR-18 / anti–IL-10 and IL-12 / IFN-β / CXCL9 armored IL- 13Rα2.BBz CAR-T cells are efficacious against orthotopic syngeneic huIL-13Rα2 + CT-2A GBM tumors in vivo. Pooled combinations of CAR-T cells were similarly evaluated against CT-2A murine glioblastoma multiforme (GBM) tumors expressing human IL-13Rα2 orthotopically in vivo. C57BL / 6J mice were implanted intracranially with 1e5 CT-2A cells. After 5 days, mice were injected intratumorally with 0.5e6 pooled CAR+ T cells (n = 5 per group). CAR-T cells were generated as described in FIG.9, except a CAR construct targeting human IL-13Rα2 was integrated. Tumor progression was tracked via in vivo imaging of radiance generated by firefly luciferase integrated into the CT-2As. The study was done in two sets (A) and (B) to cover all the combinations of interest; the IL-12 / IFN-β / CXCL9 was repeated in both studies to enable cross-comparison. Tumor radiance curves for individual mice in each treatment group are shown.

[0783] FIG.11. IL-13Rα2.BBz CAR-T cells engineered to simultaneously secrete IL-12, DR-18, and anti–IL-10 outperform those secreting IL-12, IFN-β, and CXCL9 against orthotopic syngeneic huIL-13Rα2 + CT-2A GBM tumors in vivo. IL-13Rα2.BBz CAR-T cells were engineered to secrete IL-12, DR-18, anti-IL-10, IFN-β, or CXCL9 individually, the three- output IL-12 / DR-18 / anti–IL-10 simultaneously, or the three-output IL-12 / IFN-β / CXCL9 simultaneously. C57BL / 6J mice were implanted intracranially with 1e5 CT-2A cells expressing human IL-13Rα2. After 5 days, mice were injected intratumorally with a total of 0.5e6 CAR+ T cells consisting either of a pooled population of single-output cells (“pooled”), or one population of triple-output cells (“cloned”) (n = 6 per group). Tumor progression was202666783.1 - 107 -tracked via in vivo imaging of radiance generated by firefly luciferase integrated into the CT- 2As. Rumor radiance curves for individual mice in each treatment group are shown. Results indicate the IL-12 / DR-18 / anti–IL-10 combination is superior to the IL-12 / IFN-β / CXCL9 combination. Furthermore, CAR-T cells can be engineered to simultaneously produce all three outputs and achieve effective tumor control

[0784] FIG. 12. IL-12 / DR-18 armoring is as effective as IL-12 / DR-18 / anti–IL-10 armor with IL-13Rα2.BBz CAR-T against orthotopic syngeneic muIL-13Rα2+ CT-2A GBM tumors in vivo. IL-13Rα2.BBz CAR-T cells were engineered to express IL-12, DR-18, and / or anti–IL- 10 in single-output, dual-output, or triple-output combinations. C57BL / 6J mice were implanted intracranially with 1e5 CT-2A cells expressing murine IL-13Rα2. After 5 days, mice were injected intratumorally with 0.5e6 CAR+ T cells (n = 6 per group). Tumor progression was tracked via in vivo imaging of radiance generated by firefly luciferase integrated into the CT- 2As. Tumor radiance curves for individual mice in each treatment group are shown. Results indicate IL-12 and DR-18 as a dual-output construct is sufficient to achieve robust anti-tumor efficacy.

[0785] FIG. 13A–13B. IL-12 / DR-18 armored IL-13Rα2.BBz CAR-mT are highly efficacious against orthotopic syngeneic muIL-13Rα2+ CT-2A GBM tumors in vivo. The lead combo IL-12 / DR-18 was further evaluated in vivo against the murine GBM tumor CT-2A expressing murine IL-13Rα2. C57BL / 6J mice were implanted intracranially with 1e5 CT-2A cells expressing murine IL-13Rα2. After 5 days, mice were injected intratumorally with 0.5e6 CAR+ T cells (n = 12 per group). Tumor progression was tracked via in vivo imaging of radiance generated by firefly luciferase integrated into the CT-2As. Shown here are (A) individual mouse tumor radiance values over time in each treatment group, and (B) Kaplan- Meier survival curve for each treatment group with statistics calculated by log-rank analysis, including Bonferroni correction for multiple comparisons. Results demonstrate robust anti- tumor efficacy from IL-13Rα2.BBz CAR-T cells armored with IL-12 and DR-18. Furthermore, results confirm that murine IL-13Rα2 expression does not cause the spontaneous tumor regression in negative-control groups previously observed with tumor cells expressing human IL-13Rα2.

[0786] FIG.14. IL-12 / DR-18 armoring is also effective against orthotopic syngeneic huIL- 13Ra2+ GL261 GBM tumors in vivo. The construct encoding the IL-13Ra2 CAR + IL-12 + DR-18 was further evaluated in the GL261 syngeneic GBM model. C57BL / 6J mice were implanted intracranially with 1e5 GL261 cells expressing human IL-13Rα2. After 5 days, mice were injected intratumorally with 0.5e6 IL-13Rα2 CAR+ T cells that are not armored (null-202666783.1 - 108 -output), armored with IL-12 and DR-18, or armored with IFN-γ (n = 16 per treatment group). Kaplan-Meier survival curve is shown with statistics calculated by log-rank analysis, including Bonferroni correction for multiple comparisons.

[0787] FIG.15A-15C. A phage-displayed human naïve scFv library was used to screen for phages that bind to hVEGF-A-His recombinant protein (biopanning). (A) Phage recovery from a phage-display human naïve scFv library. After the fourth round of biopanning, the recovery rate of phages had increased 25-fold over that of first round. (B) Comparison of binding by selected phage clones to human and mouse VEGF-A-His protein (mVEGF-A-His) based on ELISA at 1 × 109cfu phage titer. (C) Comparison of binding by four phage clones to hVEGF- A-His and mVEGF-A-His protein at 1 × 109cfu phage titer. The results showed that pc4-67 and pc5-76 had the two best average binding potencies for human and mouse VEGF-A-His. Ctrl-p, control phage used as a negative control; OD, optical density. Note: pc4-67 and pc5-76 correspond to VEGF 4-67 scFv and VEGF 5-76 scFv, respectively.

[0788] FIG. 16A-16E. T cells were retrovirally transduced to express anti-angiogenic CARs. Each CAR was fused to an N-terminal FLAG tag to enable surface detection by antibody staining. On day 7 of culture, transduced human T cells were stained for surface expression of FLAG-tagged (A) MSLN, (B) CLDN6, and (C) GPC CARs. On day 5 of culture, transduced murine T cells were stained for surface expression of FLAG-tagged (D) TYRP-1 and (E) IL-13Rα2 CARs. Data are mean ± 1 standard deviation with n = 3 technical replicates. Note: MSLN CARs refer to CARs utilizing the MSLN Nb C6. All data from FIG.16 onward utilize this MSLN Nb C6 CAR unless otherwise noted.

[0789] FIG. 17A–17L. VEGF 4-67 scFv expression does not impair CAR expression or antigen response of human MSLN CAR-T cells in vitro. (A) Human T cells can express and secrete the VEGF 4-67 and VEGF 5-76 scFvs. Fifteen million human CAR-T cells secreting MSLN, MSLN-Hisx6 VEGF 5-76, and MSLN-Hisx6 VEGF 4-67 were seeded on day 13 of culture. Supernatants were harvested 24 and 49 hours later and concentrated. A western blot was performed on the concentrated supernatant to probe for the poly-histidine tag. Secretion of both VEGF 5-76 and VEGF 4-67 scFvs was confirmed. VEGF 4-67 scFv was secreted at a higher level than VEGF 5-76 scFv in human T cells. (B) MSLN CAR surface expression (no antigen stimulation) in human T cells from multiple donors. Human T cells achieve similar CAR surface expression levels with or without VEGF 4-67 scFv secretion. (C–D) Human MSLN CAR-T cells show similar (C) activation marker expression and (D) tumor-cell killing upon co-incubation with MSLN+ ID8 tumor cells. (E) Human T cells expand similarly in culture upon activation with CD3 / CD28 beads regardless of VEGF 4-67 scFv secretion. Data202666783.1 - 109 -are mean ± 1 standard deviation. Paired t-test was performed on panels (B), (D), and (E). FIGS. 17F-17L are updated and include at least some of the data presented in FIGS. 17B-17E. (F) Human T cells exhibit similar CAR surface expression (%) with or without VEGF 4-67 scFv secretion but exhibit higher CAR surface expression levels (MFI). (G-H) Human MSLN CAR- T cells (G) have similar phenotypes and (H) achieve similar ex vivo T-cell fold-expansion with or without VEGF 4-67 scFv secretion. (I-K) Human MSLN CAR-T cells show similar (I) activation marker expression, (J) effector cytokine expression, and (K) tumor-cell killing upon co-incubation with MSLN+ID8 tumor cells. (L) Human MSLN CAR-T cells show similar tumor-cell killing upon co-incubation with MSLN+patient-derived PDX7 ovarian cancer cells. Data are mean ± 1 standard deviation. Each data point is representative of one independent donor for panels (F-K) and n = 6 (F, G), n = 7 (H), n = 4 (I, K), and n = 3 (J). n = 3 technical triplicates for panel (L). Two-tailed paired t-tests were performed on panels (F-H), (K), and (L). Paired one-way ANOVAs were performed on panels (I-J). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

[0790] FIG. 18A-18K. VEGF 4-67 scFv expression does not impair CAR expression or antigen response of murine MSLN CAR-T cells in vitro. (A) Murine T cells can express and secrete the VEGF 4-67 and VEGF 5-76 scFvs. Fifteen million murine CAR-T cells secreting MSLN, MSLN-Hisx6 VEGF 5-76, and MSLN-Hisx6 VEGF 4-67 were seeded on day 5 of culture. Supernatants were harvested 24 and 48 hours later and concentrated. A western blot was performed on the concentrated supernatant to probe for the poly-histidine tag. Secretion of both VEGF 5-76 and VEGF 4-67 scFvs was confirmed. VEGF 4-67 scFv was secreted at a higher level than VEGF 5-76 scFv in murine T cells. (B) Murine T cells achieve similar CAR surface expression levels with or without VEGF 4-67 scFv secretion. (C–D) Human MSLN CAR-T cells show similar (C) activation marker expression and (D) tumor-cell killing upon co-incubation with MSLN+ ID8 tumor cells. (E) Murine T cells expand similarly in culture upon activation with CD3 / CD28 beads regardless of VEGF 4-67 scFv secretion. Data are mean ± 1 standard deviation. Paired t-test was performed on panels (B), (D), and (E). FIGS. 18F- 18K are updated and include at least some of the data presented in FIGS.18B-18E. (F) Murine T cells exhibit similar CAR surface expression (%) with or without VEGF 4-67 scFv secretion but exhibit higher CAR surface expression levels (MFI). (G-H) Murine MSLN CAR-T cells (G) have similar phenotypes and (H) achieve similar ex vivo T-cell fold-expansion with or without VEGF 4-67 scFv secretion. (I-K) Murine MSLN CAR-T cells show similar (I) activation marker expression, (J) effector cytokine expression, and (K) tumor-cell killing upon co-incubation with MSLN+ID8 tumor cells. Data are mean ± 1 standard deviation. Each data202666783.1 - 110 -point is representative of one biological replicate and n = 12 (F), n = 4 (G), n = 8 (H), n = 3 (I- K). Two-tailed paired t-tests were performed on panels (F-H) and (K). Paired one-way ANOVAs were performed on panels (I-J). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. Note: “MSLN.BBz” is the same as “MSLN” CAR-T cells.

[0791] FIG. 19A-19D. VEGF-scFv 4-67 secretion significantly improves murine MSLN CAR-T cell anti-tumor efficacy against the ID8VEGF tumor. ID8VEGF is a syngeneic murine ovarian cancer model that stably overexpresses VEGF164 (mouse VEGF-A) and endogenously expresses MSLN. The tumor cells are also engineered to express firefly luciferase to facilitate tumor burden measurements via bioluminescent imaging. In the disseminated ID8VEGF tumor model, a mixed tumor dose of 0.5e6 ID8VEGF+ 8.5e6 ID8 cells (9e6 total) was injected i.p. and 3e6 CAR+ T cells were injected i.p. 6 days later. MSLN-VEGF 4-67 CAR-T cells showed improved tumor control compared to MSLN and MSLN-VEGF 5-76 CAR-T cells based on both (A) tumor burden and (B) survival. (C, D) Theses results were replicated when the CAR+ T cell dose was increased to 5e6. n = 8 (A, B) and n = 10-15 (C, D). UTD: untransduced T cells. Log-rank test was used to determine significance.

[0792] FIG.20A–20E. Bispecific MSLN / VEGF and VEGF / MSLN CAR-T cells respond to both tumor antigen and soluble VEGF. (A) Schematic of bispecific CARs. VEGF 4-67 or VEGF 5-76 scFv was used as the anti-binding domain of the CAR. (B–C) Human T cells expressing single-input VEGF CARs respond to both human and murine VEGF-A. Human T cells expressing a non-signaling transduction marker (control) or single-input VEGF CARs made with VEGF 4-67 or VEGF 5-76 scFv were cultured in the presence of exogenous human VEGF165 or murine VEGF164 for 24 hours. Surface expression of activation markers (B) CD25 and (C) CD137 were quantified by flow cytometry. (D–E) Human T cells expressing bispecific MSLN / VEGF CARs upregulate CD25 expression in response to (D) both human and murine VEGF-A and (E) MSLN+ ID8 and ID8VEGF tumor cells after a 24-hour stimulation. MFI in (D) and (E) are normalized to T-cell–only controls. TCM: tumor- conditioned media. Data in (B)–(E) are means of triplicates ± 1 standard deviation.

[0793] FIG. 21A-21F. VEGF 4-67 scFv expression does not impair CAR expression or antigen response of human IL-13Rα2 CAR-T cells in vitro. (A) Human T cells exhibit similar CAR surface expression (%) with or without VEGF 4-67 scFv secretion but exhibit higher CAR surface expression levels (MFI). (B-C) Human IL-13Rα2 CAR-T cells (B) have similar phenotypes and (C) achieve similar ex vivo T-cell fold-expansion with or without VEGF 4-67 scFv secretion. (D-F) Human IL-13Rα2 CAR-T cells show similar (D) activation marker expression, (E) effector cytokine expression, and (F) tumor-cell killing upon co-incubation202666783.1 - 111 -with IL-13Rα2+CT-2A tumor cells. Data are mean ± 1 standard deviation. Each data point is representative of one independent donor and n = 4 (A-C) and n = 3 (D-F). Two-tailed paired t- tests were performed on panels (A-C) and (F). Paired one-way ANOVAs were performed on panels (D-E). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

[0794] FIG. 22A-22F. VEGF 4-67 scFv expression does not impair CAR expression or antigen response of murine IL-13Rα2 CAR-T cells in vitro. (A) Murine T cells exhibit similar CAR surface expression (%) with or without VEGF 4-67 scFv secretion but exhibit higher CAR surface expression levels (MFI). (B-C) Murine IL-13Rα2 CAR-T cells (B) have similar phenotypes and (C) achieve similar ex vivo T-cell fold-expansion with or without VEGF 4-67 scFv secretion. (D-F) Murine IL-13Rα2 CAR-T cells show similar (D) activation marker expression, (E) effector cytokine expression, and (F) tumor-cell killing upon co-incubation with IL-13Rα2+CT-2A tumor cells. Data are mean ± 1 standard deviation. Each data point is representative of one biological replicate and n = 11 (A), n = 5 (B), n = 8 (C), and n = 3 (D-F). Two-tailed paired t-tests were performed on panels (A-C) and (F). Paired one-way ANOVAs were performed on panels (D-E). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

[0795] FIG. 23A–23B. VEGF4-67 -scFv secretion significantly improves murine IL13- Rα2 CAR-T cell anti-tumor efficacy against the CT-2A tumor. CT-2A is a syngeneic murine glioma tumor model. In this experiment, the CT-2A cells were engineered to overexpress the murine IL13-Rα2 antigen, and engineered to express firefly luciferase to facilitate tumor burden measurements via bioluminescent imaging. In this tumor model, 1e5 CT-2A tumor cells were injected intracranially (i.c.) and 5e5 CAR+ T cells were injected i.c. 6 days later. As a negative control, T cells were engineered to express a truncated epidermal growth factor receptor (EGFRt) incapable of targeting the CT-2A tumor cells. Anti-VEGF scFv secretion improves the anti-tumor efficacy of murine IL13Rα2 CAR-T cells based on both (A) tumor burden and (B) survival. n = 12 for EGFRt and IL13Rα2, and n = 8 for IL13Rα2-VEGF 4-67. Log-rank test was used to determine significance.

[0796] FIG.24. VEGF 4-67 scFv secretion results in reduced VEGF concentration in vivo. C57BL / 6 mice were injected i.c. with 1e5 CT-2A tumor cells. Six days later, tumor-bearing mice were either left untreated or injected i.c. with 5e5 murine T cells expressing the indicated construct. Two days after CAR-T cell injection, mice were euthanized and brains were harvested. These brains were enzymatically and mechanically digested to achieve a single-cell suspension. The remaining supernatant was analyzed via ELISA to quantify VEGF concentrations. Anti-VEGF scFv secretion by T cells reduces VEGF concentrations in the brains of CT-2A tumor bearing mice. Data is mean ± 1 standard deviation. n = 4 for every202666783.1 - 112 -treatment. One-way ANOVA was performed. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

[0797] FIG. 25A-25E. VEGF 4-67 scFvs can modulate CT-2A tumor vasculature. C57BL / 6 mice were injected i.c. with 1e5 CT-2A tumor cells. Nine days later, tumor-bearing mice were injected i.c. with 5e5 murine T cells expressing the indicated construct. Two days after CAR-T cell injection, mice were euthanized and brains were harvested and prepared for confocal microscopy. (A-B) Representative images of CT-2A vasculature as visualized by CD31 staining and either (A) VEGF or (B) pimonidazole (hypoxia marker) staining. Scale bar = 50 μm. (C-E) Quantifications of (C) % CD31+ area (D) % VEGF+ area, and (E) % pimonidazole+ area show that CAR-T cell treatment significantly increases intratumoral vessel density, VEGF expression, and hypoxia, whereas aberrations in each of these measures were corrected by anti-VEGF scFv secretion, indicating that VEGF 4-67 scFvs can prevent excessive tumor angiogenesis and modulate tumor vessels. Bars show population mean ± 1 standard deviation. Each data point represents the mean value from 3 regions of each individual brain section, and multiple sections were taken from each of 3 brains per treatment. n = 18 for panel (C), with 6 sections from each of 3 brains. n = 9 for panels (D) and (E), with 3 sections from each of the 3 brains. One-way ANOVAs were performed on panels (C-E). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

[0798] FIG. 26. VEGF 4-67 scFv secretion significantly reduces peripheral myeloid cell infiltration into CT-2A tumor-bearing brains that occurs in response to murine IL-13Rα2 CAR- T cell treatment. C57BL / 6 mice were injected i.c. with 1e5 CT-2A tumor cells. Six days later, tumor-bearing mice were either left untreated or injected i.c. with 5e5 murine T cells expressing the indicated construct. Two days after CAR-T cell injection, mice were euthanized and brains were harvested. These brains were enzymatically and mechanically digested to achieve a single-cell suspension and analyzed via flow cytometry to quantify the number of monocytes and macrophages per brain. Data are mean ± 1 standard deviation. n = 3 for every treatment. One-way ANOVA was performed. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

[0799] FIG. 27A-D. VEGF 4-67 scFv secretion skews CD8 T cells towards highly proliferative and chemotactic subtypes, and remodels monocytes and endothelial cells to be more pro-inflammatory. C57BL / 6 mice were injected i.c. with 1e5 CT-2A tumor cells. Six days later, tumor-bearing mice were injected i.c. with either PBS 5e5 murine T cells expressing the indicated construct. A control group of “naïve” mice that were given neither tumor nor T cells was also included. Two days after CAR-T cell injection, mice were euthanized and brains were harvested. These brains were enzymatically and mechanically digested to achieve a202666783.1 - 113 -single-cell suspension and subjected to single-cell RNA sequencing. (A) Distribution of CD8 T-cell subtypes. In both EGFRt and IL-13Rα2 CAR-T cells, VEGF 4-67 scFv secretion en...

Claims

WHAT IS CLAIMED:

1. A chimeric polypeptide comprising: I. (a) an antigen binding domain comprising a nanobody, wherein the nanobody comprises a complementarity determining region (CDR) 1, CDR2, and CDR3 having at least 80% sequence identity to the CDR1, CDR2, CDR3 from the nanobody of SEQ ID NO:57; (b) a transmembrane domain; and (c) an intracellular signaling domain; II (a) an antigen binding domain comprising a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:36 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:37; (b) a transmembrane domain; and (c) an intracellular signaling domain; III. (a) an antigen binding domain comprising a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:45 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:46; (b) a transmembrane domain; and (c) an intracellular signaling domain; or IV. (a) an antigen binding domain comprising a nanobody, wherein the nanobody comprises a complementarity determining region (CDR) 1, CDR2, and CDR3 having at least 80% sequence identity to the CDR1, CDR2, CDR3 from the nanobody of SEQ ID NO:53;202666783.1 - 243 -(b) a transmembrane domain; and (c) an intracellular signaling domain.

2. The chimeric polypeptide of claim 1, wherein: I. the nanobody comprises a CDR1, CDR2, and CDR3 having the amino acid sequence of the CDR1, CDR2, and CDR3, respectively, from the nanobody of SEQ ID NO:57; II. the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:36 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:37; III. the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:45 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:46; or IV. the nanobody comprises a CDR1, CDR2, and CDR3 having the amino acid sequence of the CDR1, CDR2, and CDR3, respectively, from the nanobody of SEQ ID NO:

53.

3. The chimeric polypeptide of claim 1 or 2, wherein: I. the CDR1, CDR2, and CDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:58-60, respectively; II. the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:38-43, respectively; III. the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:47-52, respectively; or IV. the CDR1, CDR2, and CDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:54-56, respectively.

4. The chimeric polypeptide of claim 3, wherein:202666783.1 - 244 -I. the CDR1, CDR2, and CDR3 each comprise the amino acid sequence of SEQ ID NOS:58-60, respectively; II. the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:38-43, respectively; III. the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:47-52, respectively; or IV. the CDR1, CDR2, and CDR3 each comprise the amino acid sequence of SEQ ID NOS:54-56, respectively.

5. The chimeric polypeptide of any one of claims 1-4, wherein: I. the antigen binding domain and / or nanobody comprises the amino acid sequence of SEQ ID NO:57 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:57; II. the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:36 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:37; or the VH comprises the amino acid sequence of SEQ ID NO:36 and / or the VL comprises the amino acid sequence of SEQ ID NO:37; III. the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:45 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:46; or the VH comprises the amino acid sequence of SEQ ID NO:45 and / or the VL comprises the amino acid sequence of SEQ ID NO:46; or IV. the antigen binding domain and / or nanobody comprises the amino acid sequence of SEQ ID NO:53 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:

53.

6. A polypeptide comprising the amino acid sequence of any one of SEQ ID NOS:138- 207 or an amino acid sequence having at least 80% sequence identity to the amino acid sequences of SEQ ID NOS:138-207.

7. An anti-VEGF antibody or antigen binding fragment comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein: (a) the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:2 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1,202666783.1 - 245 -LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:3; or (b) the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:19 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:

20.

8. The antibody or antigen binding fragment of claim 7, wherein: (a) the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:2 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:3; or (b) the VH comprises a HCDR1, HCDR2, and HCDR3 having the amino acid sequence of a HCDR1, HCDR2, and HCDR3, respectively, from a VH comprising the amino acid sequence of SEQ ID NO:19 and wherein the VL comprises a LCDR1, LCDR2, and LCDR3 having the amino acid sequence of the LCDR1, LCDR2, and LCDR3, respectively, from a VL comprising the amino acid sequence of SEQ ID NO:

20.

9. The antibody or antigen binding fragment of claim 7 or 8, wherein: (a) the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:4-9, respectively; or the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:4-9, respectively; or (b) the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:21-26, respectively; or the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:21-26, respectively.

10. The antibody or antigen binding fragment of any one of claims 7-9, wherein: (a) the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:2 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:3; or the VH comprises the amino acid sequence of SEQ ID NO:2 and / or the VL comprises the amino acid sequence of SEQ ID NO:3; or202666783.1 - 246 -(b) the VH comprises an amino acid sequence with at least 80% sequence identity to SEQ ID NO:19 and / or the VL comprises an amino acid sequence with at least 80% sequence identity to SEQ ID NO:20; or the VH comprises the amino acid sequence of SEQ ID NO:19 and / or the VL comprises the amino acid sequence of SEQ ID NO:

20.

11. A polypeptide comprising the antigen binding fragment of any one of claims 7-10.

12. The polypeptide of claim 11, wherein the polypeptide: (i) comprises at least two antigen binding fragments, wherein each antigen binding fragment is independently selected from an antigen binding fragment of any one of claims 7- 11; (ii) the polypeptide is multivalent; and / or (iii) the polypeptide is bispecific.

13. A chimeric polypeptide comprising: (a) an antigen binding fragment of any one of claims 7-11; (b) a transmembrane domain; and (c) an intracellular signaling domain.

14. The chimeric polypeptide of claim 13, wherein the antigen binding fragment comprises SEQ ID NO:1 or SEQ ID NO:18 or an amino acid sequence having at least 70% sequence identity to SEQ ID NO:1 or SEQ ID NO:

18.

15. The chimeric polypeptide of any one of claims 1-5 or 14, further comprising a signal peptide and optionally wherein the signal peptide comprises SEQ ID NO:122 or 123 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:122 or 123.

16. The chimeric polypeptide of any one of claims 1-5 or 14-15, wherein: (i) the transmembrane domain is an α or β chain of the T cell receptor or a transmembrane domain from CD28, CD3-epsilon, CD45, CD4, CD5, CD8-α, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD123, CD134, CD137 or CD154; optionally wherein the transmembrane domain is a CD28 transmembrane domain; and / or (ii) the transmembrane domain comprises SEQ ID NO:130 or 131 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:130 or 131; and / or (iii) the intracellular signaling domain comprises a primary signaling domain; optionally wherein the primary signaling domain is a CD3ζ (zeta) signaling domain; optionally wherein the primary signaling domain comprises SEQ ID NO:134 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:134; and / or (iv) the intracellular signaling domain further comprises a costimulatory domain; optionally wherein the costimulatory domain comprises a signaling domain from 4-1BB202666783.1 - 247 -(CD137), CD28, IL-15Rα, OX40, CD2, CD27, CDS, ICAM-1, LFA-1 (CD11a / CD18), or ICOS (CD278); optionally wherein the costimulatory domain comprises a CD28 or 4-1BB signaling domain; optionally wherein the costimulatory domain comprises SEQ ID NO:132 or 133 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:132 or 133; (v) . the antigen binding domain and the transmembrane domain are separated by a hinge region; optionally wherein the hinge region comprises an IgG4 hinge, a CD8α hinge, an IgG1 hinge, or a CD34 hinge; optionally wherein the hinge region is between 8 and 300 amino acids in length; optionally wherein the hinge region comprises one of SEQ ID NOS:126-129 or an amino acid sequence having at least 80% sequence identity to one of SEQ ID NOS:126- 129.

17. A fusion protein comprising the antigen binding fragment of any one of claims 7-11 and a CD3 binding region and optionally wherein the CD3 binding region comprises a CD3- specific fragment antigen binding (Fab), single chain variable fragment (scFv), single domain antibody, or single chain antibody.

18. One or more nucleic acid(s) comprising a sequence encoding the polypeptide, protein, antigen, or antigen binding fragment of any of claims 1-17.

19. A nucleic acid comprising a sequence encoding: I. (i) a chimeric antigen receptor (CAR) or engineered T-cell receptor (TCR); and (ii) an IL-12 polypeptide; and / or (iii) an IL-18 polypeptide; or II. (i) a chimeric antigen receptor (CAR) or engineered T-cell receptor (TCR); and (ii) an IL-12 polypeptide; (iii) an IFN-β polypeptide; and / or (iv) a CXCL9 polypeptide.

20. The nucleic acid of claim 18 or 19, wherein: (a) the nucleic acid further encodes an IL-12 polypeptide and an IL-18 polypeptide; and / or (b) the nucleic acid further encodes an IL-12, IFN-β, and / or CXCL9 polypeptide.

21. The nucleic acid(s) of any one of claims 18-20, wherein: (a) the IL-12 polypeptide comprises a p40 and / or p35 subunit; and / or (b) the IL-12 polypeptide comprises a single-chain IL-12 polypeptide; and / or202666783.1 - 248 -(c) the p40 subunit is amino-proximal to the p35 subunit or the p35 subunit is amino- proximal to the p40 subunit; and / or (d) the nucleic acid further encodes for a suicide gene product and optionally wherein the suicide gene product is herpes simplex virus thymidine kinase (HSV-TK), purine nucleoside phosphorylase (PNP), cytosine deaminase (CD), carboxypetidase G2, cytochrome P450, linamarase, β-lactamase, nitroreductase (NTR), carboxypeptidase A, split Cas9, truncated EGFR, or inducible caspase 9; and / or (e) the nucleic acid comprises one or more promoter(s) that direct the expression of the suicide gene product, CAR, IL-12 polypeptide, IFN-β polypeptide, CXCL9 polypeptide, and / or IL-18 polypeptide. (f) the IL-12, IFN-β, CXCL9, and / or IL-18 polypeptide is derived from a human IL-12, human IFN-β, human CXCL9, and / or human IL-18 polypeptide; and / or (g) the IL-18 polypeptide comprises a decoy resistant IL-18 polypeptide; and / or (h) the p40 polypeptide comprises the amino acid sequence of SEQ ID NO:236 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:236; and / or (i) the p35 polypeptide comprises the amino acid sequence of SEQ ID NO:237 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:237; and / or (j) the IL-12 polypeptide comprises the amino acid sequence of SEQ ID NO:230 or 234 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:230 or 234; and / or (k) the IL-18 polypeptide comprises the amino acid sequence of SEQ ID NO:229, 232, or 238-240 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:229, 232, or 238-240; and / or (l) the IFN-β polypeptide comprises the amino acid sequence of SEQ ID NO:253 or 255 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:253 or 255; and / or (m) the CXCL9 polypeptide comprises the amino acid sequence of SEQ ID NO:257 or 259 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:257 or 259; and / or (n) the nucleic acid encodes for a signal sequence at the amino-terminus of the IL-12, IFN-β, CXCL9 and / or IL-18 polypeptide and optionally wherein the signal sequence comprises SEQ ID NO:122, 123, 235, 241, 254, 256, 258, and / or 260; and / or202666783.1 - 249 -(o) the IL-12 polypeptide and the IL-18 polypeptide are expressed from one nucleic acid or the IL-12 polypeptide, IFN-β polypeptide, and the CXCL9 polypeptide are expressed from one nucleic acid; and / or (p) the nucleic acid encodes for a self-cleaving peptide or internal ribosome entry sequence (IRES), wherein the self-cleaving peptide or IRES is between the IL-12, IL-18, CXCL9, and / or IFN-β polypeptides and optionally wherein the self-cleaving polypeptide comprises SEQ ID NO:135 or 136.

22. The nucleic acid of any one of claims 18-21, wherein the nucleic acid encodes for a chimeric antigen receptor (CAR), wherein the CAR comprises: (a) an antigen binding domain; (b) a transmembrane domain; and (c) an intracellular signaling domain; and wherein the antigen binding domain comprises an anti-TYRP-1, anti-IL-13Rα2, anti-TGF-β, anti-CLDN6, anti-GPC3, or anti-MSLN antigen binding domain.

23. The nucleic acid of claim 22, wherein the antigen binding domain comprises an anti- IL-13Rα2 comprising an IL-13 mutein polypeptide and optionally wherein the IL-13 mutein polypeptide comprises the amino acid sequence of SEQ ID NO:115 or 233, or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:115 or 233.

24. The nucleic acid of claim 22, wherein the antigen binding domain comprises anti- MSLN and optionally wherein the antigen binding domain comprises: (a) a nanobody, wherein the nanobody comprises a complementarity determining region (CDR) 1, CDR2, and CDR3 from the nanobody of SEQ ID NO:57 or a CDR1, CDR2, and CDR3 each having at least 80% sequence identity to the CDR1, CDR2, CDR3 from the nanobody of SEQ ID NO:57; (b) a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 from SEQ ID NO:36 or a HCDR1, HCDR2, and HCDR3 each having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:36 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 from a VL of SEQ ID NO:37 or a LCDR1, LCDR2, and LCDR3 each having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:37; (c) a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and202666783.1 - 250 -HCDR3 from the VH of SEQ ID NO:45 or a HCDR1, HCDR2, and HCDR3 each having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:45 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 from the VL of SEQ ID NO:46 or a LCDR1, LCDR2, and LCDR3, each having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:46; or (d) a nanobody, wherein the nanobody comprises a complementarity determining region (CDR) 1, CDR2, and CDR3 from the nanobody of SEQ ID NO:53 or a CDR1, CDR2, and CD3 having at least 80% sequence identity to the CDR1, CDR2, CDR3 from the nanobody of SEQ ID NO:

53.

25. The nucleic acid of claim 24, wherein: (a) the CDR1, CDR2, and CDR3 each comprise the amino acid sequence of SEQ ID NOS:58-60, respectively or the CDR1, CDR2, and CDR3 have at least 80% sequence identity to SEQ ID NOS:58-60, respectively; (b) the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:38-43, respectively or an amino acid sequence that has at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:38-43, respectively; (c) the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:47-52, respectively or an amino acid sequence that has at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:47-52, respectively; or (d) the CDR1, CDR2, and CDR3 each comprise the amino acid sequence of SEQ ID NOS:54-56, respectively or or an amino acid sequence that has at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:54-56, respectively.

26. The nucleic acid of claim 22, wherein the antigen binding domain comprises anti- TYRP-1 and wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 from a VH having the amino acid sequence of SEQ ID NO:62 or having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:62 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1,202666783.1 - 251 -LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:63 or having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:63; and optionally wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:64-69, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:64-69, respectively; and / or wherein the VH comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:62 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:63, or wherein the VH comprises the amino acid sequence of SEQ ID NO:62 and / or the VL comprises the amino acid sequence of SEQ ID NO:

63.

27. The nucleic acid of claim 22, wherein the antigen binding domain comprises anti-TGF- β; and wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein: (a) the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 from a VH having the amino acid sequence of SEQ ID NO:71 or having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:71 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:72 or having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:72; (b) the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 from a VH having the amino acid sequence of SEQ ID NO:80 or having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:80 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:81 or having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:81; or (c) the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 from a VH having the amino acid sequence of SEQ ID NO:89 or having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:89 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 from a VL having202666783.1 - 252 -the amino acid sequence of SEQ ID NO:90 or having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:

90.

28. The nucleic acid of claim 27, wherein: (a) the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:73-78, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:73-78, respectively; (b) the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:82-87, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:82-87, respectively; or (c) wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:91- 96, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:91-96, respectively.

29. The nucleic acid of claim 22, wherein the antigen binding domain comprises anti- CLDN6 and wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 from a VH having the amino acid sequence of SEQ ID NO:98 or a HCDR1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:98 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:99 or a LCDR1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:99; and optionally wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:100-103, the sequence TSN, and SEQ ID NO:105, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:100-103, the sequence TSN, and SEQ ID NO:105, respectively.

30. The nucleic acid of claim 22, wherein the antigen binding domain comprises anti-GPC3 and wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 from a VH having the amino acid sequence of SEQ202666783.1 - 253 -ID NO:107 or a HCDR1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:107 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:108 or a LCDR1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:108; and optionally the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:109-114, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:109- 114, respectively.

31. The nucleic acid of any one of claims 24-30, wherein: (a) the VH is amino proximal to the VL or the VL is amino proximal to the VH; and / or (b) the antigen binding domain comprises a linker between the VH and VL; optionally wherein the linker comprises a glycine serine linker; optionally wherein the linker comprises (G4S)n, wherein n is an integer selected from 1-10; and / or optionally wherein the linker comprises or consists of SEQ ID NO:119, 120, or 121; and / or (c) the antigen binding domain comprises the amino acid sequence of one of SEQ ID NOS:35, 44, 53, 57, 61, 70, 79, 88, 97, or 106 or an amino acid sequence having at least 80% sequence identity to one of SEQ ID NOS:35, 44, 53, 57, 61, 70, 79, 88, 97, or 106; and / or (d) the nucleic acid is DNA or is RNA; and / or (e) the nucleic acid further comprises a signal peptide; optionally wherein the signal peptide comprises SEQ ID NO:122 or 123 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:122 or 123; and / or (f) the transmembrane domain is an α or β chain of the T cell receptor or a transmembrane domain from CD28, CD3-epsilon, CD45, CD4, CD5, CD8-α, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD123, CD134, CD137 or CD154; optionally wherein the transmembrane domain is a CD28 transmembrane domain; optionally wherein the transmembrane domain comprises SEQ ID NO:130 or 131 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:130 or 131; and / or (g) the intracellular signaling domain comprises a primary signaling domain; optionally wherein the primary signaling domain is a CD3ζ (zeta) signaling domain; optionally wherein202666783.1 - 254 -the primary signaling domain comprises SEQ ID NO:134 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:134; and / or (h) wherein the intracellular signaling domain further comprises a costimulatory domain; optionally wherein the costimulatory domain comprises a signaling domain from 4- 1BB (CD137), CD28, IL-15Rα, OX40, CD2, CD27, CDS, ICAM-1, LFA-1 (CD11a / CD18), or ICOS (CD278); optionally wherein the costimulatory domain comprises a CD28 or 4-1BB signaling domain; and optionally wherein the costimulatory domain comprises SEQ ID NO:132 or 133 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:132 or 133; and / or (i) wherein the antigen binding domain and the transmembrane domain are separated by a hinge region; optionally wherein the hinge region comprises an IgG4 hinge, a CD8α hinge, an IgG1 hinge, or a CD34 hinge; optionally wherein the hinge region is between 8 and 300 amino acids in length; optionally wherein the hinge region comprises one of SEQ ID NOS:126- 129 or an amino acid sequence having at least 80% sequence identity to one of SEQ ID NOS:126-129.

32. A nucleic acid comprising a sequence encoding a polypeptide comprising the amino acid sequence of one of SEQ ID NOS:138-207 or an amino acid sequence having at least 80% sequence identity to one of SEQ ID NOS:138-207.

33. A polypeptide expressed from the nucleic acid of any one of claims 18-32.

34. A cell or population of cells comprising the nucleic acid of any of claims 18-32; optionally wherein the cell expresses the nucleic acid of any one of claims 18-32.

35. A cell or population of cells comprising: (a) a heterologous nucleic acid encoding an IL-12 polypeptide and a heterologous nucleic acid encoding an IL-18 polypeptide; or (b) a heterologous nucleic acid encoding an IL-12 polypeptide, a heterologous nucleic acid encoding an IFN-β polypeptide and a heterologous nucleic acid encoding a CXCL9 polypeptide.

36. The cell(s) of claim 34 or 35, wherein: (a) the cell is a T cell, a natural killer (NK) cell, a natural killer T cell (NKT), an invariant natural killer T cell (iNKT), stem cell, lymphoid progenitor cell, peripheral blood mononuclear cell (PBMC), bone marrow cell, fetal liver cell, embryonic stem cell, cord blood cell, induced pluripotent stem cell (iPS cell), naïve memory T cell, CD4+ T cell, and / or CD8+ T cell; and / or202666783.1 - 255 -(b) the cell comprises a T cell from a population of CD14 depleted, CD25 depleted, and / or CD62L enriched PBMCs; and / or (c) the population comprises 103-108cells.

37. The cell(s) of any one of claims 34-36, wherein the cell(s) further comprises: (a) a heterologous nucleic acid encoding an IL-12 polypeptide and a heterologous nucleic acid encoding an IL-18 polypeptide; or (b) a heterologous nucleic acid encoding an IL-12 polypeptide, a heterologous nucleic acid encoding an IFN-β polypeptide and a heterologous nucleic acid encoding a CXCL9 polypeptide.

38. The cell(s) of any one of claims 34-37 wherein: (a) the IL-12 polypeptide comprises a p40 and / or p35 subunit; and / or (b) the IL-12 polypeptide comprises a single-chain IL-12 polypeptide; and / or (c) the p40 subunit is amino-proximal to the p35 subunit or the p35 subunit is amino- proximal to the p40 subunit; and / or (d) the IL-12 polypeptide comprises a linker between the p40 and p35 subunit; and / or (e) the nucleic acid encoding the IL-12, IFN-β, CXCL9 and / or IL-18 polypeptide is an expression construct; optionally wherein the expression construct is a plasmid or viral vector; optionally wherein the viral vector is a vector derived from a retrovirus or a vector derived from a lentivirus; and / or (f) the cell comprises a nucleic acid encoding for a suicide gene product; optionally wherein the suicide gene product is herpes simplex virus thymidine kinase (HSV-TK), purine nucleoside phosphorylase (PNP), cytosine deaminase (CD), carboxypetidase G2, cytochrome P450, linamarase, β-lactamase, nitroreductase (NTR), carboxypeptidase A, split Cas9, truncated EGFR, or inducible caspase 9; and / or (g) wherein the nucleic acid comprises one or more promoter(s) that direct the expression of the suicide gene product, IL-12, IFN-β, CXCL9, and / or IL-18 polypeptide; optionally wherein the promoter is constitutive or conditional; optionally wherein the promoter responds positively to at least one cytokine or to T-cell activation; optionally wherein the at least one cytokine is TGF- β, IL-6, TNF α, IFN-γ, IL-1β, IL-2, IL-8, or IL-10 or the promoter responds positively to NFAT or NF-κB; and / or (h) the IL-12, IFN- β, CXCL9, and / or IL-18 polypeptide is derived from a human IL- 12, human IFN- β, human CXCL9, and / or human IL-18 polypeptide; optionally wherein the IL-18 polypeptide comprises a decoy resistant IL-18 polypeptide; and / or202666783.1 - 256 -(i) the p40 polypeptide comprises the amino acid sequence of SEQ ID NO:236 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:236; and / or (j) the p35 polypeptide comprises the amino acid sequence of SEQ ID NO:237 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:237; and / or (k) the IL-12 polypeptide comprises the amino acid sequence of SEQ ID NO:230 or 234 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:230 or 234; and / or (l) the IL-18 polypeptide comprises the amino acid sequence of SEQ ID NO:229, 232, or 238-240 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:229, 232, or 238-240; and / or (m) the IFN-β polypeptide comprises the amino acid sequence of SEQ ID NO:253 or 255 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:253 or 255; and / or (n) the CXCL9 polypeptide comprises the amino acid sequence of SEQ ID NO:257 or 259 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:257 or 259; and / or (o) the nucleic acid encodes for a signal sequence at the amino-terminus of the IL-12, IFN-β, CXCL9 and / or IL-18 polypeptide; optionally wherein the signal sequence comprises SEQ ID NO:122, 123, 235, 241, 254, 256, 258, and / or 260; and / or (p) the IL-12 polypeptide, the IL-18 polypeptide, IFN-β polypeptide, CXCL9 polypeptide, and / or anti-VEGF antigen binding fragment are expressed from one nucleic acid; and / or (q) the nucleic acid expresses a polypeptide comprising the IL-12 polypeptide, IL-18 polypeptide, IFN-β polypeptide, CXCL9 polypeptide, and / or anti-VEGF antigen binding fragment and / or wherein the IL-12 polypeptide, IL-18 polypeptide, IFN-β polypeptide, CXCL9 polypeptide, and / or anti-VEGF antigen binding fragment are expressed from one nucleic acid; and / or (r) the nucleic acid encodes for a self-cleaving peptide or internal ribosome entry sequence (IRES), wherein the self-cleaving peptide or IRES is between the IL-12 polypeptide, IL-18 polypeptide, IFN-β, polypeptide, CXCL9 polypeptide, and / or anti-VEGF antigen binding fragment; optionally wherein the self-cleaving polypeptide comprises SEQ ID NO:135 or 136; and / or (s) the cell further comprises a nucleic acid encoding a CAR or TCR.202666783.1 - 257 -39. The cell(s) of claim 38, wherein the cells comprise a nucleic acid encoding a CAR and wherein the the CAR comprises: (a) an antigen binding domain; (b) a transmembrane domain; and (c) an intracellular signaling domain; wherein the antigen binding domain comprises an anti-TYRP-1, anti-IL-13Rα2, anti-TGF-β, anti-CLDN6, anti-GPC3, or anti-MSLN antigen binding domain.

40. The cell(s) of claim 39, wherein the antigen binding domain comprises an anti-IL- 13Rα2 comprising an IL-13 mutein polypeptide and optionally wherein the IL-13 mutein polypeptide comprises the amino acid sequence of SEQ ID NO:115 or 233, or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:115 or 233.

41. The cell(s) of claim 39, wherein the antigen binding domain comprises anti-MSLN and optionally wherein the antigen binding domain comprises: (a) a nanobody, wherein the nanobody comprises a complementarity determining region (CDR) 1, CDR2, and CDR3 from the nanobody of SEQ ID NO:57 or a CDR1, CDR2, and CDR3 each having at least 80% sequence identity to the CDR1, CDR2, CDR3 from the nanobody of SEQ ID NO:57; (b) a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 from SEQ ID NO:36 or a HCDR1, HCDR2, and HCDR3 each having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:36 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 from a VL of SEQ ID NO:37 or a LCDR1, LCDR2, and LCDR3 each having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:37; (c) a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 from the VH of SEQ ID NO:45 or a HCDR1, HCDR2, and HCDR3 each having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:45 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 from the VL of SEQ ID NO:46 or a LCDR1, LCDR2, and LCDR3, each having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:46; or202666783.1 - 258 -(d) a nanobody, wherein the nanobody comprises a complementarity determining region (CDR) 1, CDR2, and CDR3 from the nanobody of SEQ ID NO:53 or a CDR1, CDR2, and CD3 having at least 80% sequence identity to the CDR1, CDR2, CDR3 from the nanobody of SEQ ID NO:

53.

42. The cell(s) of claim 41, wherein: (a) the CDR1, CDR2, and CDR3 each comprise the amino acid sequence of SEQ ID NOS:58-60, respectively or the CDR1, CDR2, and CDR3 have at least 80% sequence identity to SEQ ID NOS:58-60, respectively; (b) the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:38-43, respectively or an amino acid sequence that has at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:38-43, respectively; (c) the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:47-52, respectively or an amino acid sequence that has at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:47-52, respectively; or (d) the CDR1, CDR2, and CDR3 each comprise the amino acid sequence of SEQ ID NOS:54-56, respectively or or an amino acid sequence that has at least 80% sequence identity to the amino acid sequence of SEQ ID NOS:54-56, respectively.

43. The cell(s) of claim 39, wherein the antigen binding domain comprises anti-TYRP-1 and wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 from a VH having the amino acid sequence of SEQ ID NO:62 or having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:62 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:63 or having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:63; and optionally wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:64-69, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:64-69, respectively; and / or wherein the VH comprises an amino acid sequence with at least 70%202666783.1 - 259 -sequence identity to SEQ ID NO:62 and / or the VL comprises an amino acid sequence with at least 70% sequence identity to SEQ ID NO:63, or wherein the VH comprises the amino acid sequence of SEQ ID NO:62 and / or the VL comprises the amino acid sequence of SEQ ID NO:

63.

44. The cell(s) of claim 39, wherein the antigen binding domain comprises anti-TGF-β and wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein: (a) the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 from a VH having the amino acid sequence of SEQ ID NO:71 or having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:71 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:72 or having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:72; (b) the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 from a VH having the amino acid sequence of SEQ ID NO:80 or having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:80 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:81 or having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:81; or (c) the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 from a VH having the amino acid sequence of SEQ ID NO:89 or having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:89 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:90 or having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:

90.

45. The cell(s) of claim 44, wherein: (a) the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:73-78, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:73-78, respectively;202666783.1 - 260 -(b) the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:82-87, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:82-87, respectively; or (c) wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:91- 96, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:91-96, respectively.

46. The cell(s) of claim 39, wherein the antigen binding domain comprises anti-CLDN6 and wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 from a VH having the amino acid sequence of SEQ ID NO:98 or a HCDR1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:98 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:99 or a LCDR1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:99; and optionally wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:100- 103, the sequence TSN, and SEQ ID NO:105, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:100-103, the sequence TSN, and SEQ ID NO:105, respectively.

47. The cell(s) of claim 39, wherein the antigen binding domain comprises anti-GPC3 and wherein the antigen binding domain comprises a variable heavy chain (VH) and a variable light chain region (VL), wherein the VH comprises a heavy chain complementarity determining region (HCDR) 1, HCDR2, and HCDR3 from a VH having the amino acid sequence of SEQ ID NO:107 or a HCDR1, HCDR2, and HCDR3 having at least 80% sequence identity to the HCDR1, HCDR2, HCDR3 from a VH having the amino acid sequence of SEQ ID NO:107 and wherein the light chain variable region comprises a light chain complementarity determining region (LCDR) 1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:108 or a LCDR1, LCDR2, and LCDR3 having at least 80% sequence identity to the LCDR1, LCDR2, and LCDR3 from a VL having the amino acid sequence of SEQ ID NO:108; and optionally the antigen binding domain comprises a variable heavy chain (VH) and a202666783.1 - 261 -variable light chain region (VL), wherein the VH comprises a HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 having an amino acid sequence that has at least 80% sequence identity to SEQ ID NOS:109-114, respectively or wherein the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 each comprise the amino acid sequence of SEQ ID NOS:109- 114, respectively.

48. The cell(s) of any one of claims 41-47, wherein: (a) the VH is amino proximal to the VL or the VL is amino proximal to the VH; and / or (b) the antigen binding domain comprises a linker between the VH and VL; optionally wherein the linker comprises a glycine serine linker; optionally wherein the linker comprises (G4S)n, wherein n is an integer selected from 1-10; and / or optionally wherein the linker comprises or consists of SEQ ID NO:119, 120, or 121; and / or (c) the antigen binding domain comprises the amino acid sequence of one of SEQ ID NOS:35, 44, 53, 57, 61, 70, 79, 88, 97, or 106 or an amino acid sequence having at least 80% sequence identity to one of SEQ ID NOS:35, 44, 53, 57, 61, 70, 79, 88, 97, or 106; and / or (d) the nucleic acid further comprises a signal peptide; optionally wherein the signal peptide comprises SEQ ID NO:122 or 123 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:122 or 123; and / or (e) the transmembrane domain is an α or β chain of the T cell receptor or a transmembrane domain from CD28, CD3-epsilon, CD45, CD4, CD5, CD8-α, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD123, CD134, CD137 or CD154; optionally wherein the transmembrane domain is a CD28 transmembrane domain; optionally wherein the transmembrane domain comprises SEQ ID NO:130 or 131 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:130 or 131; and / or (f) the intracellular signaling domain comprises a primary signaling domain; optionally wherein the primary signaling domain is a CD3ζ (zeta) signaling domain; optionally wherein the primary signaling domain comprises SEQ ID NO:134 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:134; and / or (g) wherein the intracellular signaling domain further comprises a costimulatory domain; optionally wherein the costimulatory domain comprises a signaling domain from 4- 1BB (CD137), CD28, IL-15Rα, OX40, CD2, CD27, CDS, ICAM-1, LFA-1 (CD11a / CD18), or ICOS (CD278); optionally wherein the costimulatory domain comprises a CD28 or 4-1BB signaling domain; and optionally wherein the costimulatory domain comprises SEQ ID NO:132 or 133 or an amino acid sequence having at least 80% sequence identity to SEQ ID NO:132 or 133; and / or202666783.1 - 262 -(h) the antigen binding domain and the transmembrane domain are separated by a hinge region; optionally wherein the hinge region comprises an IgG4 hinge, a CD8α hinge, an IgG1 hinge, or a CD34 hinge; optionally wherein the hinge region is between 8 and 300 amino acids in length; optionally wherein the hinge region comprises one of SEQ ID NOS:126-129 or an amino acid sequence having at least 80% sequence identity to one of SEQ ID NOS:126-129; (i) the cells are a population of cells comprising 103-1013cells.

49. A composition comprising the polypeptide of any one of claims 1-6, 11-16, or 33, the anti-VEGF antibody or antigen binding fragment of any one of claims 7-10, the protein of claim 17, nucleic acids of any one of claims 18-32, or cells of any one of claims 34-48, wherein the composition is a pharmaceutically acceptable formulation.

50. A method of making a cell comprising introducing into a cell the nucleic acid of any of claims 18-32 into a cell.

51. A method of treating a patient with cancer or for increasing anti-tumor immunity and / or tumor cell cytotoxicity in a patient having cancer, the method comprising administering to the patient an effective amount of the composition of claim 49.

52. The method of claim 51, wherein the patient has and / or has been determined to have a MSLN+ cancer.

53. The method of claim 51 or 52, wherein the cancer comprise ovarian cancer, stomach cancer, colorectal cancer, endometrial cancer, uterine cancer, acute myeloid leukemia, pancreatic cancer, lung cancer, mesothelioma, melanoma, glioma, glioblastoma, sarcoma, hepatocellular carcinoma, lung cancer, non-small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, breast cancer, neuroblastoma, thyroid cancer, gastric cancer, testis cancer, or uterine cancer.

54. The method of any one of claims 51-53, further comprising administering an additional therapy to the patient, optionally wherein the additional therapy comprises an immunotherapy, anti-angiogenic therapy, chemotherapy, surgery, radiotherapy, neoantigen therapy, or vaccination; optionally wherein the additional therapy comprises immunotherapy and wherein the immunotherapy comprises immune checkpoint inhibitor therapy or bispecific T cell engagers.

55. The method of any one of claims 51-54, wherein the cells are autologous.202666783.1 - 263 -

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