EP4569091A1 - Hypoimmunogenic modified cells - Google Patents

Abstract

The present disclosure relates to modified cells, e.g., chimeric antigen receptor (CAR) T cells and/or exogenous T cell receptor (eTCR) cells, which have reduced in vivo immunogenicity, and the use thereof.

Description

HYPOIMMUNOGENIC MODIFIED CELLS

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority benefit of U.S. Provisional Application No. 63/371,060, filed August 10, 2022, which is incorporated by reference herein in its entirety.

REFERENCE TO SEQUENCE LISTING SUBMITTED

ELECTRONICALLY VIA EFS-WEB

[0002] The content of the electronically submitted sequence listing (Name:

3817_134PC01_SequenceListing_ST26.xml, Size: 69,591 bytes; and Date of Creation: August 9, 2023) submitted in this application is incorporated herein by reference in its entirety.

FIELD

[0003] The present disclosure relates to modified cells, e.g., chimeric antigen receptor (CAR) immune cells and/or exogenous T cell receptor (eTCR) immune cells, that have reduced in vivo immunogenicity, and the uses thereof.

BACKGROUND

[0004] Cell based immunotherapy is a rapidly developing field of research for the development of novel and improved methods of treating various diseases, including cancer. Various therapies have been approved comprising isolating T cells from a subject, modifying the T cells to express a chimeric antigen receptor (CAR) or an exogenous T cell receptor (eTCR) that is capable of targeting the T cell to a particular antigen, and administering the modified T cells back to the same subject. Though autologous CAR-T or eTCR therapy has proven to be quite promising, this process can be time consuming and expensive, with inconsistent results at least in part due to donor variability. [0005] One means of streamlining the process is by creating "off-the-shelf¹ modified immune cells, which are derived from induced pluripotent stem cells (iPSCs). However, administering iPSC-derived immune cells that are not HLA-matched to the patient will likely lead to an immune response in the patient, resulting in the patient rejecting the cell therapy. As such, there remains a need in the art for developing iPSC-derived cell therapies that can evade a patient's immune response.

BRIEF SUMMARY

[0006] The present disclosure provides, at least in part, novel methods and modified cells that are capable of evading killing by a patient’s immune system, when the cells (e.g., a population of immune cells or induced pluripotent cells transduced with a CAR or a TCR and containing certain modifications described herein) are administered to the patient as a cell therapy product. The present invention is based, at least in part, on the discovery that reducing expression of endogenous MHC-I HLAs and MHC-II HLAs, while expressing a Fas dominant negative (Fas-DN) and an HLA-E in cells (such as, for example, immune cells) results in cells that are surprisingly able to have longer persistence in vivo not only by evading natural killer (NK) cell-mediated killing and T cell-mediated killing but also by avoiding Fas-mediated apoptosis. Such a combination of modifications helps to clear a major hurdle of off-the-shelf immune cell therapies.

[0007] Some aspects of the present disclosure are directed to a modified cell, comprising: (i) a chimeric antigen receptor (CAR) and/or an exogenous T cell receptor (eTCR); (ii) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; and (iv) a polypeptide comprising a dominant-negative Fas (Fas-DN), a Fas-CD27 chimeric polypeptide (Fas-CD27), a Fas-4-lBB chimeric polypeptide (Fas-BB), a Fas-OX40 chimeric polypeptide (Fas-OX40), or any combination thereof.

[0008] In some aspects, the MHC-I human leukocyte antigens are HLA- A, HLA-B and HLA-C. In some aspects, the MHC-II human leukocyte antigens are HLA-DP, HLA-DQ and HLA-DR. In some aspects, the reduced expression of the MHC-I human leukocyte antigens results from a mutation in, or a deletion of, one or more endogenous genes encoding beta-2-microglobulin (B2M). In some aspects, the reduced expression of the endogenous MHC-II human leukocyte antigens results from a mutation in, or a deletion of, the one or more endogenous genes encoding class II major histocompatibility complex transactivator (CIITA).

[0009] In some aspects, the cell further comprises a reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type. In some aspects, the reduced expression of the CD58 results from a mutation in, or a deletion of, one or more endogenous genes encoding CD58. In some aspects, the cell comprises a nucleic acid encoding Fas-DN. In some aspects, the cell comprises a nucleic acid encoding Fas-CD27. In some aspects, the cell comprises a nucleic acid encoding Fas-BB. In some aspects, the cell comprises a nucleic acid encoding Fas-OX40.

[0010] Some aspects of the present disclosure are directed to a modified cell comprising: (i) a CAR or an eTCR; (ii) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; and (iv) a Fas-DN.

[0011] Some aspects of the present disclosure are directed to a cell comprising: (i) a CAR or an eTCR; (ii) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; and (iv) a Fas-CD27.

[0012] Some aspects of the present disclosure are directed to a cell comprising: (i) a CAR or an eTCR; (ii) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; and (iv) a Fas-BB.

[0013] Some aspects of the present disclosure are directed to a cell comprising: (i) a CAR or an eTCR; (ii) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; and (iv) a Fas-OX40.

[0014] In some aspects, the cell further comprises a reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type. In some aspects, the reduced expression of the CD58 results from, a mutation in, or a deletion of, one or more endogenous genes encoding CD58.

[0015] In some aspects, the cell further comprises a reduced expression of endogenous poliovirus receptor (PVR) relative to a wild-type cell of the same cell type. In some aspects, the reduced expression of the PVR results from a mutation in, or a deletion of, one or more endogenous genes encoding PVR.

[0016] In some aspects, the cell further comprises an increased expression of HLA-E relative to a wild-type cell of the same cell type. In some aspects, the increased expression of the HLA-E results from one of: (i) introducing into the cells, a nucleic acid encoding an HLA-E polypeptide; and (ii) modifying an endogenous gene encoding an HLA-E polypeptide, wherein the modification increases the expression of the endogenous HLA-E polypeptide; or (iii) both (i) and (ii). In some aspects, the HLA-E polypeptide is a chimeric polypeptide comprising an HLA-E polypeptide linked to a B2M polypeptide.

[0017] In some aspects, the method further comprises transfecting the cell with a nucleic acid encoding an interleukin 15 (IL15) polypeptide. In some aspects, the IL15 polypeptide is a membrane bound IL15/IL15Ralpha fusion polypeptide (mIL15/Ra).

[0018] In some aspects, the method further comprises introducing into the cell a heterologous nucleic acid encoding a human chemokine (C-C motif) ligand 19 (CCL19) polypeptide.

[0019] In some aspects, the cell is an immune cell, an induced pluripotent stem cell (iPSC) or a cell differentiated from iPSC. In some aspects, the cell is an immune cell or a hematopoietic stem cell differentiated from an iPSC. In some aspects, the cell comprises a T cell, an NK cell, an NKT cell, or a tumor-infiltrating lymphocyte.

[0020] In some aspects, the CAR or the eTCR comprises an antigen-binding domain that specifically binds a tumor antigen. In some aspects, the tumor antigen comprises CD 19, CD20, ROR1, CD22, carcinoembryonic antigen, alphafetoprotein, CA-125, 5T4, MUC-1, epithelial tumor antigen, prostate-specific antigen, melanoma-associated antigen, mutated p53, mutated ras, HER2/Neu, folate binding protein, HIV-1 envelope glycoprotein gpl20, HIV-1 envelope glycoprotein gp41, GD2, CD123, CD33, CD138, CD23, CD30, CD56, c- Met, mesothelin, GD3, HERV-K, IL-llRalpha, kappa chain, lambda chain, CSPG4, ERBB2, EGFRvIII, VEGFR2, HER2-HER3 in combination, HER1-HER2 in combination, NY-ESO-1, synovial sarcoma X breakpoint 2 (SSX2), melanoma antigen (MAGE), melanoma antigen recognized by T cells 1 (MART-1), gplOO, prostate specific antigen (PSA), prostate specific membrane antigen (PSMA), prostate stem cell antigen (PSCA), GPC3, BCMA, GCC, ADGRE, claudin or any combination thereof.

[0021] Some aspects of the present disclosure are directed to a cell prepared according to a method disclosed herein. In some aspects, the cell has increased persistence in vivo relative to a wild-type cell of the same cell type.

[0022] Some aspects of the present disclosure are directed to a population of cells comprising a cell disclosed herein.

[0023] Some aspects of the present disclosure are directed to a cell population, wherein at least 50% of the cells in the population comprises a cell disclosed herein.

[0024] Some aspects of the present disclosure are directed to a method of treating a subject in need thereof, comprising administering to the subject a cell or a population of cells disclosed herein.

[0025] In some aspects, the subject is afflicted with a cancer. In some aspects, the cancer comprises bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, Hodgkin's Disease, non-Hodgkin's lymphoma, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, chronic or acute leukemia, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, solid tumors of childhood, lymphocytic lymphoma, cancer of the bladder, cancer of the kidney or ureter, carcinoma of the renal pelvis, neoplasm of the central nervous system (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid cancer, squamous cell cancer, T-cell lymphoma, environmentally induced cancers including those induced by asbestos, or any combination thereof.

[0026] Some aspects of the present disclosure are directed to a guide RNA capable of hybridizing to a human CD58 gene, comprising a nucleic acid sequence selected from SEQ ID NOs: 1-20 and 41. In some aspects, the guide RNA comprises the nucleic acid sequence set forth in SEQ ID NO: 1. In some aspects, the guide RNA consists of the nucleic acid sequence set forth in SEQ ID NO: 1. In some aspects, the guide RNA comprises the nucleic acid sequence set forth in SEQ ID NO: 2. In some aspects, the guide RNA consists of the nucleic acid sequence set forth in SEQ ID NO: 2. In some aspects, the guide RNA comprises the nucleic acid sequence set forth in SEQ ID NO: 3. In some aspects, the guide RNA consists of the nucleic acid sequence set forth in SEQ ID NO: 3. In some aspects, the guide RNA comprises the nucleic acid sequence set forth in SEQ ID NO: 4. In some aspects, the guide RNA consists of the nucleic acid sequence set forth in SEQ ID NO: 4.

[0027] Some aspects of the present disclosure are directed to a method of inactivating a human CD58 gene in a cell, comprising contacting the cell with a CD58 guide RNA disclosed herein or a nucleic acid encoding the guide RNA and a DNA endonuclease or a nucleic acid encoding the DNA endonuclease. In some aspects, the DNA endonuclease comprises CRISPR/Cas9.

[0028] Some aspects of the present disclosure are directed to a dominant-negative Fas (Fas-DN), comprising an amino acid sequence set forth in SEQ ID NO: 27.

[0029] Some aspects of the present disclosure are directed to a Fas-CD27 chimeric polypeptide (Fas-CD27), comprising an amino acid sequence set forth in SEQ ID NO: 22.

[0030] Some aspects of the present disclosure are directed to a Fas-4-lBB chimeric polypeptide (Fas-4-lBB), comprising an amino acid sequence set forth in SEQ ID NO: 23.

[0031] Some aspects of the present disclosure are directed to a Fas-OX40 chimeric polypeptide (Fas-OX40), comprising an amino acid sequence set forth in SEQ ID NO: 24, SEQ ID NO: 25, or SEQ ID NO: 26.

[0032] In some aspects, the eTCR is a gamma-delta TCR. In some aspects, the gammadelta TCR is V gamma 9-V delta 2 TCR (g9d2TCR).

[0033] Some aspects of the present disclosure are directed to a modified cell comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; and (iii) a nucleic acid encoding a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type, a nucleic acid encoding a suicide gene or any combination thereof.

[0034] Some aspects of the present disclosure are directed to a modified cell comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); and (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof.

[0035] Some aspects of the present disclosure are directed to a modified cell comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; and (iv) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof.

[0036] Some aspects of the present disclosure are directed to a method of engineering a human cell, comprising: (i) inactivating one or more endogenous genes encoding MHC-I human leukocyte antigens, wherein the inactivation results in a reduced expression of MHC-I human leukocyte antigens relative to a wild-type cell of the same cell type; (ii) inactivating one or more endogenous genes encoding MHC-II human leukocyte antigens, wherein the inactivation results in a reduced expression of MHC-II human leukocyte antigens relative to a wild-type cell of the same cell type; and (iii) transfecting the cell with a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof.

[0037] Some aspects of the present disclosure are directed to a method of engineering a human cell, comprising: (i) inactivating one or more endogenous genes encoding MHC-I human leukocyte antigens, wherein the inactivation results in a reduced expression of MHC-I human leukocyte antigens relative to a wild-type cell of the same cell type; (ii) inactivating one or more endogenous genes encoding MHC-II human leukocyte antigens, wherein the inactivation results in a reduced expression of MHC-II human leukocyte antigens relative to a wild-type cell of the same cell type; (iii) inactivating one or more endogenous genes encoding CD58, wherein the inactivation results in a reduced expression of CD58 relative to a wild-type cell of the same cell type; and (iv) transfecting the cell with a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof. BRIEF DESCRIPTION OF THE DRAWINGS/FIGURES

[0038] FIGs. 1A-1D provide cell surface phenotype (CTV-CD25+) comparison of HLA mis-matched peripheral blood mononuclear cells ("PBMC"; FIGs. 1A-1B) and HLA matched PBMCs (FIGs. 1C-1D) that were co-cultured with either wild-type ("WT") meso induced chimeric antigen receptor ("iCAR")-T cells (FIGs. 1A and 1C) or MHC-I/MHC- II double knock-out ("dKO") meso iCAR-T (FIGs. IB and ID), as analyzed by flow cytometry. FIG. IE is a bar graph showing the percent of CD25+ dividing T cells from either HLA-matching donor or HLA-mismatch donor after co-culture with WT meso iCAR-T or MHC-I/MHC-II dKO meso iCAR-T cells , as indicated.

[0039] FIG. 2 is a bar graph showing the cytotoxic effect of alloreactive ("allo")-T cells on meso iCAR-T cells that were (i) edited (WT); (ii) B2M/CIITA double KO; (iii) HLA- ABC/CIITA double KO; (iv) or HLA-ABC/CIITA double KO that further over-express HLA-G and HLA-E. Cells were incubated at 2: 1 and 1 : 1 effectortarget (ET) ratios for 24 hours, as indicated.

[0040] FIGs. 3A-3D are line graphs illustrating the effects of NK cell-mediated killing of meso iCAR-T cells. NK cells were obtained from four donors (donor 1 : FIGs. 3A-3D; donor 2: FIGs. 3E-3H, donor 3: FIGs. 3I-3L; and donor 4: FIGs. 3M-3P). meso iCAR-T cells were labeled with carbonxyfluorescein diacetate succinimidyl ester (CFSE; Life Technologies) prior to co-incubation with NK cells, meso iCAR-T cells were DKO (FIGs. 3 A, 3E, 31, and 3M); DKO and PVR knock-out (FIGs. 3B, 3F, 3 J, and 3N); DKO and CD58 knock-out (FIGs. 3C, 3G, 3K, and 30); and DKO, PVR knock-out, and CD58 knock-out (FIGs. 3D, 3H, 3L, and 3P). Cells were incubated at a ratio of 5: 1, 2: 1, and 0: 1 effector: target (E:T) ratio for 24 hours. Percent dead cells were evaluated by FACS and analyzed using FlowJo software. "DKO" refers to B2M/CIITA double knock-out.

[0041] FIG. 4 is a line graph illustrating the effects of NK cell-mediated killing of meso iCAR-T cells, meso iCAR-T cells were labeled with carbonxyfluorescein diacetate succinimidyl ester (CFSE; Life Technologies) prior to co-incubation with NK cell lines at a ratio of 4: 1, 2: 1, and 1 : 1 effector: target (E:T) ratio for 24 hours. Percent dead meso iCAR-T cells were evaluated by FACS and analyzed using FlowJo software. "DKO" refers to B2M/CIITA double knock-out. "TKO" refers to B2M/CIITA/CD58 triple knockout. “+E” referes to further overexpression of HLA-E. “+F” referes to further expression of FasDN. “+FasBB” refers to further expression of Fas-41BB switch receptor. “+HSVTK” refers to further expression of HSVTK.

[0042] FIGs. 5A-5C are line graphs illustrating the effects of NK cell-mediated killing of meso iCAR-T cells. NK cells were obtained from three donors (donor 1 : FIG. 5 A; donor 2: FIG. 5B; and donor 3: FIG. 5C). meso iCAR-T cells were labeled with carbonxyfluorescein diacetate succinimidyl ester (CFSE; Life Technologies) prior to coincubation with NK cell lines at a ratio of 5: 1, and 1.5: 1 effector: target (E:T) ratio for 24 hours. Percent dead meso iCAR-T cells were evaluated by FACS and analyzed using FlowJo software.

[0043] FIG. 6 is a line graph illustrating the tumor cell killing effects of meso iCAR-T cells, meso iCAR-T cells and GSU tumor cells were co-cultured at a ratio of 10: 1, 3: 1, 1 : 1 and 0.3 : 1 effector: target (E:T) ratio for 24 hours. The number of viable cells were assessed by Cell Titer-Gio Luminescent Viability Assay which results in cell lysis and generation of a luminescent signal proportional to the amount of ATP present. The luminescent signal was measured using the Pherastar plate reader.

[0044] FIGs. 7A-7C are line graphs showing NK cell-mediated killing of meso iCAR-T cells over time. FIG. 7A shows the normalized time course where NSG-MHC I/II DKO animals were first dosed with 1.5xl0⁶ NK cells and then dosed with WT (Wild Type, positive control), B2M and CIITA KO (untransduced or UTD), B2M/CIITA KO with overexpression of HLA-E and FasDN, HLA-ABC KO with overexpression of HLA-G, or HLA-ABC knock-out with overexpression of HLA-E and FasDN iCAR T cells. iCAR T cells were administered at a dose of IxlO⁶ cells per animal. FIGs. 7B-7C show normalized time course data where NSG-MHC Eli DKO animals were first dosed with 2.0xl0⁶ NK cells from two donors (FIG. 7B and FIG. 7C). After 1 hour the animals were dosed with WT (Wild Type), B2M/CIITA KO (untransduced or UTD), B2M/CIITA KO with overexpression of HLA-E and FasDN, B2M/CIITA/PVR KO with overexpression of HLA-E and FasDN, B2M/CIITA/CD58 KO with overexpression of HLA-E and FasDN, or B2M/CIITA/CD58/PVR KO with overexpression of HLA-E and FasDN iCART cells at a dose of 1x10⁶ cells per animal.

[0045] FIGs. 8A-8D are line graphs illustrating the persistence of iCART cells over time. FIG. 8A shows NK cell-mediated killing of iCART cells further modified to overexpress FasBB and/or the HSVTK kill switch. NSG-MHC Eli DKO animals were first dosed with 2.0xl0⁶ NK cells. After 1 hour, the animals were dosed with WT (Wild Type); B2M/CIITA KO (untransduced or UTD; DKO-UTD); B2M/CIITA/PVR KO (TKO- UTD); B2M/CIITA/PVR KO (TKO) with overexpression of HLA-E and FasDN; B2M/CIITA/CD58 KO (TKO) with overexpression of HLA-E and HSVTK; B2M/CIITA/ PVR KO (TKO) with overexpression of HLA-E, FasDN, and HSVTK; B2M/CIITA/ PVR KO (TKO) with overexpression of HLA-E, FasBB, and HSVTK; wherein each iCART is administered at a dose of IxlO⁶ cells per animal. FIG. 8B shows data from an alloreactive T cell rejection assay. NSG-MHC Eli DKO mice were intraperitoneally given 200 uL of freshly prepared alloreactive T cells. One hour later, mice were administered 200 uL of iCAR T cells (WT, DKO-UTD, DKO with overexpression of HLA-E and FASDN; DKO with PVR KO and overexpression of HLA-E and FASDN; DKO with CD58 KO and overexpression of HLA-E and FASDN; or DKO with CD58 and PVR KO, and overexpression of HLA-E and FASDN) was administered. Animals were imaged on Day 1, Day 3, and Day 6 on the IVIS Spectrum for a bioluminescent signal. This signal was then quantified as total flux (photons/second) within the region of interest (ROI). FIG. 8C-8D illustrate GSU tumor cell killing efficacy, wherein NSG- MHC I/II DKO mice were first given IxlO⁶ luciferized GSU cells followed by WT, B2M/ CIITA KO, B2M/ CIITA KO with overexpresson of HLA-E and FasDN, or B2M/CIITA/CD58 KO with overexpression of HLA-E and FasDN, at a dose of IxlO⁶ cells per animal.

[0046] FIGS. 9A-9B are schematic representations of the use of a Fas receptor in modified immune cells to affect apoptosis.

[0047] FIG. 10A-10F are graphical representations of cell proliferation of cells expressing mIL15/Ra (5BB) (FIG. 10A), dnFAS (FIG. 10B), FAS-4-1BB (FIG. 10C), and FAS-OX40 (FIG. 10D) following stimulation with FAS super ligand. FIG. 10E is a bar graph showing the fold change in cell counts following addition of super FAS-ligand at 0.1 ng/ml, 1 ng/ml, 10 ng/ml, and 100 ng/ml. FIG. 10F shows fold expansion during activation in iCAR T cells and cells transduced with 5BB, dnFAS, FAS-4-1BB, and FAS- 0X40 constructs.

[0048] FIGs. 11 A-l IB are line graphs illustrating cell proliferation (FIG. 11 A) and percent cell lysis of target cells (FIG. 1 IB) for (i) untransduced cells prepared with boost and non-boost protocols (UTD, UTD-Boost), (ii) iCAR T cells expressing 5BB prepared with boost and non-boost protocols (5BB, 5BB-Boost), (iii) dnFAS/5BB, and (iv) FAS- OX40/5BB. [0049] FIGs. 12A-12C are line graphs showing in vivo efficacy data of modified iCART cells in a mouse model. GSU-RFluc cells were intraperitoneally introduced to NSG-MHC I/II DKO mice at a dose of IxlO⁶ cells per animal. On day 4, iCAR-T cells were introduced at a dose of IxlO⁶ cells per animal and cell lysis was analyzed based on mean fluorescence intensity. FIG. 12A shows cell lysis over time in 7 study groups (PBS, UTD+IL-15, BBp+IL-15, BB-FasDN+IL-15, BB-Fas/41BB+IL-15, BB-Fas/CD27+IL- 15, and BB-Fas/Ox40+IL-15). FIGs. 12B-12C show individual animal responses to PBS vs 5BB (FIG. 12B) FAS-Ox40 comparison (FIG. 12C).

[0050] FIGs. 13A-13F are graphical representations of flow cytometry of HLA-E expression in HLA-E knock-in iPSCs (FIG. 13 A) and wild-type cells (FIG. 13B), FASDN expression in FASDN knock-in iPSCs (FIG. 13C) and wild-type cells (FIG. 13D), and tEGFR expression in tEGFR knock-in iPSCs (FIG. 13E) and wild-type cells (FIG. 13F).

DETAILED DESCRIPTION

[0051] Some aspects of the present disclosure are directed to modified cells, comprising: (i) a chimeric antigen receptor (CAR) and/or an exogenous T cell receptor (eTCR); (ii) reduced expression of MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; (iii) reduced expression of MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; and (iv) a polypeptide comprising a dominant-negative Fas (Fas-DN), a Fas-CD27 chimeric polypeptide (Fas-CD27), a Fas- 4-1BB chimeric polypeptide (Fas-BB), a Fas-OX40 chimeric polypeptide (Fas-OX40), or any combination thereof.

[0052] Some aspects of the present disclosure are directed to modified cells comprising: (i) a CAR or an eTCR; (ii) reduced expression of MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; (iii) reduced expression of MHC- II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; and (iv) a Fas-DN.

[0053] Some aspects of the present disclosure are directed to modified cells comprising: (i) a CAR or an eTCR; (ii) reduced expression of MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; (iii) reduced expression of MHC- II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; and (iv) a Fas-CD27.

[0054] Some aspects of the present disclosure are directed to modified cells comprising: (i) a CAR or an eTCR; (ii) reduced expression of MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; (iii) reduced expression of MHC- II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; and (iv) a Fas-BB.

[0055] Some aspects of the present disclosure are directed to modified cells comprising: (i) a CAR or an eTCR; (ii) reduced expression of MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; (iii) reduced expression of MHC- II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; and (iv) a Fas-OX40.

[0056] In some aspects, the MHC-I human leukocyte antigens are HLA- A, HLA-B and HLA-C. In some aspects, the reduced expression of the MHC-I human leukocyte antigens or inactivated endogenous gene encoding MHC-I human leukocyte antigens results from a mutation in, insertion (knock-in) of a transgene to or a deletion (knock-out) of one or more endogenous genes encoding beta-2-microglobulin (B2M). In some aspects, the reduced expression of the MHC-I human leukocyte antigens or inactivated endogenous gene encoding MHC-I human leukocyte antigens results from an insertion (knock-in) of a transgene encoding HLA-E or protein comprising HLA-E as a whole or in part to one or more endogenous genes encoding beta-2-microglobulin (B2M). In some aspects, a transgene encoding HLA-E or protein comprising HLA-E as a whole or in part may be inserted to the site of exon 1, 2 or 3 of the endogenous gene encoding beta-2- microglobulin (B2M), preferably to the site of exon 1. In some aspects, the MHC-II human leukocyte antigens are HLA-DP, HLA-DQ and HLA-DR. In some aspects, the reduced expression of the endogenous MHC-II human leukocyte antigens or inactivated endogenous gene encoding MHC-II human leukocyte antigens results from a mutation in, insertion of a transgene into or a deletion (knock-out) of the one or more endogenous genes encoding class II major histocompatibility complex transactivator (CIITA). In some aspects, the reduced expression of the MHC-II human leukocyte antigens or inactivated endogenous gene encoding MHC-II human leukocyte antigens results from an insertion (knock-in) of a transgene encoding a Fas (e.g., Fas-DN), a suicide gene (e.g., HSV-TK or the variant thereof) and/or tag protein (e.g., EGFR, LNGFR or the variant thereof) as a whole or in part to one or more endogenous genes encoding CIITA. In some aspects, a transgene encoding a Fas (e.g., Fas-DN), a suicide gene (e.g., HSV-TK or the variant thereof) and/or tag protein (e.g., EGFR, LNGFR or the variant thereof) as a whole or in part may be inserted to the site of exon 1, 2 or 3 of the endogenous gene encoding CIITA, preferably to the site of exon 3. The transgene may comprise genes encoding one or more proteins comprising a Fas (e.g., Fas-DN), a suicide gene (e.g., HSV-TK or the variant thereof) and/or tag protein (e.g., EGFR, LNGFR or the variant thereof) as a whole or in part. In some aspects, the reduced expression of CD58 or inactivated endogenous gene encoding CD58 results from a mutation in, insertion (knock-in) of a transgene to or a deletion (knock-out) of one or more endogenous genes encoding CD58. The site of a deletion (knock-out) of one or more endogenous genes encoding CD58 can be exon 1, 2 and/or 3 of the endogenous gene encoding CD58, preferably exon 3.

[0057] Before the present disclosure is described in greater detail, it is to be understood that this disclosure is not limited to the particular compositions or process steps described, as such can, of course, vary. As will be apparent to those of skill in the art upon reading this disclosure, each of the individual aspects described and illustrated herein has discrete components and features which can be readily separated from or combined with the features of any of the other several aspects without departing from the scope or spirit of the present disclosure. Any recited method can be carried out in the order of events recited or in any other order that is logically possible.

[0058] The headings provided herein are not limitations of the various aspects of the disclosure, which can be defined by reference to the specification as a whole. It is also to be understood that the terminology used herein is for the purpose of describing particular aspects only, and is not intended to be limiting.

I. Terms

[0059] In order that the present disclosure can be more readily understood, certain terms are first defined. As used in this application, except as otherwise expressly provided herein, each of the following terms shall have the meaning set forth below. Additional definitions are set forth throughout the application.

[0060] As described herein, any concentration range, percentage range, ratio range or integer range is to be understood to include the value of any integer within the recited range and, when appropriate, fractions thereof (such as one tenth and one hundredth of an integer), unless otherwise indicated.

[0061] Throughout this disclosure, the term "a" or "an" entity refers to one or more of that entity; for example, "a chimeric polypeptide," is understood to represent one or more chimeric polypeptides. As such, the terms "a" (or "an"), "one or more," and "at least one" can be used interchangeably herein.

[0062] Furthermore, "and/or" where used herein is to be taken as specific disclosure of each of the two specified features or components with or without the other. Thus, the term "and/or" as used in a phrase such as "A and/or B" herein is intended to include "A and B," "A or B," "A" (alone), and "B" (alone). Likewise, the term "and/or" as used in a phrase such as "A, B, and/or C" is intended to encompass each of the following aspects: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone). In addition, "or" is used mean an open list of the components in the list. For example, "wherein X comprises A or B" means X comprises A, X comprises B, X comprises A and B, or X comprises A or B and any other components.

[0063] The terms "about" or "comprising essentially of' refer to a value or composition that is within an acceptable error range for the particular value or composition as determined by one of ordinary skill in the art, which will depend in part on how the value or composition is measured or determined, /.< ., the limitations of the measurement system. For example, "about" or "comprising essentially of can mean within 1 or more than 1 standard deviation per the practice in the art. Alternatively, "about" or "comprising essentially of can mean a range of up to 10%. Furthermore, particularly with respect to biological systems or processes, the terms can mean up to an order of magnitude or up to 5-fold of a value. When particular values or compositions are provided in the application and claims, unless otherwise stated, the meaning of "about" or "comprising essentially of should be assumed to be within an acceptable error range for that particular value or composition.

[0064] The terms "activated immune cells," "activated T cells," and "activated NK cells" refer to, among other things, immune cells, e.g., T cells and/or NK cells, that are undergoing cell division.

[0065] An "antigen" refers to any molecule, e.g., a peptide, that provokes an immune response or is capable of being bound by a TCR. The immune response may involve antibody production, the activation of specific immunologically-competent cells, or a combination thereof. A person of skill in the art would readily understand that any macromolecule, including virtually all proteins or peptides, can serve as an antigen. An antigen can be endogenously expressed, i.e. expressed by genomic DNA, or can be recombinantly expressed. An antigen and/or an epitope can be specific to a certain tissue, such as a cancer cell, or it can be broadly expressed. In addition, fragments of larger molecules can act as antigens. In one aspect, antigens are tumor antigens.

[0066] An "antigen-presenting cell" or "APC," as used herein, refers to a cell or a celllike antigen-presenting surface that expresses one or more antigen. In some aspects, the antigen is displayed on the surface of the APC.

[0067] An "anti -turn or effect" as used herein, refers to a biological effect that can present as a decrease in tumor volume, a decrease in the number of tumor cells, a decrease in tumor cell proliferation, a decrease in the number of metastases, an increase in overall or progression-free survival of a patient, an increase in life expectancy of a patient, or amelioration of various physiological symptoms in a patient associated with the tumor. An anti-tumor effect can also refer to the prevention of the occurrence of a tumor, e.g., a vaccine.

[0068] As used herein, the term "approximately," as applied to one or more values of interest, refers to a value that is similar to a stated reference value. In certain aspects, the term "approximately" refers to a range of values that fall within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value).

[0069] The term "autologous" refers to any material, e.g., an immune cell, derived from the same individual to which it is later to be re-introduced. For example, an autologous T cell therapy comprises administering to a subject a T cell that was isolated from the same subject. The term "allogeneic" refers to any material derived from one individual which is then introduced to another individual of the same species. For example, an allogeneic T cell transplantation comprises administering to a subject a T cell that was obtained from a donor other than the subject.

[0070] A "cancer" refers to a broad group of various diseases characterized by the uncontrolled growth of abnormal cells in the body. Unregulated cell division and growth results in the formation of malignant tumors that invade neighboring tissues and may also metastasize to distant parts of the body through the lymphatic system or bloodstream. A "cancer" or "cancer tissue" can include a tumor. Examples of cancers that can be treated by the methods of the present invention include, but are not limited to, cancers of the immune system including lymphoma, leukemia, and other leukocyte malignancies. In some embodiments, the methods of the present invention can be used to reduce the tumor size of a tumor derived from, for example, the cancer comprises bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, breast cancer, prostate cancer, lung cancer (e.g. , non-small cell lung cancer (NSCLC) or small cell lung cancer (SCLC)), Hodgkin's Disease, nonHodgkin's lymphoma, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, chronic or acute leukemia, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, solid tumors of childhood, lymphocytic lymphoma, cancer of the bladder, cancer of the kidney or ureter, carcinoma of the renal pelvis, neoplasm of the central nervous system (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid cancer, squamous cell cancer, T-cell lymphoma, environmentally induced cancers including those induced by asbestos, or any combination thereof. The particular cancer can be responsive to chemo- or radiation therapy or the cancer can be refractory. A refractory cancer refers to a cancer that is not amendable to surgical intervention, and the cancer is either initially unresponsive to chemo- or radiation therapy or the cancer becomes unresponsive over time.

[0071] It is understood that wherever aspects are described herein with the language "comprising," otherwise analogous aspects described in terms of "consisting of and/or "consisting essentially of' are also provided.

[0072] A "cytokine," as used herein, refers to a non-antibody protein that is released by one cell in response to contact with a specific antigen, wherein the cytokine interacts with a second cell to mediate a response in the second cell. A cytokine can be endogenously expressed by a cell, added to a cell in culture, administered to a subject, or any combination thereof. Cytokines may be released by immune cells, including macrophages, B cells, T cells, and mast cells to propagate an immune response. Cytokines can induce various responses in the recipient cell. Cytokines can include homeostatic cytokines, chemokines, pro-inflammatory cytokines, effectors, and acutephase proteins. For example, homeostatic cytokines, including interleukin (IL) 7 and IL- 15, promote immune cell survival and proliferation, and pro-inflammatory cytokines can promote an inflammatory response. Examples of homeostatic cytokines include, but are not limited to, IL-2, IL-4, IL-5, IL-7, IL-10, IL-12p40, IL-12p70, IL-15, IL-21, and interferon (IFN) gamma. Examples of pro-inflammatory cytokines include, but are not limited to, IL-la, IL-lb, IL-6, IL-13, IL-17a, tumor necrosis factor (TNF)-alpha, TNF- beta, fibroblast growth factor (FGF) 2, granulocyte macrophage colony-stimulating factor (GM-CSF), soluble intercellular adhesion molecule 1 (sICAM-1), soluble vascular adhesion molecule 1 (sVCAM-1), vascular endothelial growth factor (VEGF), VEGF-C, VEGF-D, and placental growth factor (PLGF). Examples of effectors include, but are not limited to, granzyme A, granzyme B, soluble Fas ligand (sFasL), and perforin. Examples of acute phase-proteins include, but are not limited to, C-reactive protein (CRP) and serum amyloid A (SAA).

[0073] Chemokines" are a type of cytokine that mediates cell chemotaxis, or directional movement. Examples of chemokines include, but are not limited to, IL-8, IL- 16, eotaxin, eotaxin-3, macrophage-derived chemokine (MDC or CCL22), monocyte chemotactic protein 1 (MCP-1 or CCL2), MCP-4, macrophage inflammatory protein la (MIP-la, MIP- la), MIP-Ib (MIP-lb), gamma-induced protein 10 (IP- 10), and thymus and activation regulated chemokine (TARC or CCL17).

[0074] Other examples of cytokines include, but are not limited to chemokine (C-C motif) ligand (CCL) 1, CCL5, monocyte-specific chemokine 3 (MCP3 or CCL7), monocyte chemoattractant protein 2 (MCP-2 or CCL8), CCL 13, IL-1, IL-3, IL-9, IL-11, IL- 12, IL- 14, IL- 17, IL-20, IL-21, granulocyte colony- stimulating factor (G-CSF), leukemia inhibitory factor (LIF), oncostatin M (OSM), CD 154, lymphotoxin (LT) beta, 4- IBB ligand (4-1BBL), a proliferation-inducing ligand (APRIL), CD70, CD153, CD 178, glucocorticoid-induced TNFR-related ligand (GITRL), tumor necrosis factor superfamily member 14 (TNFSF14), OX40L, TNF- and ApoL-related leukocyte- expressed ligand 1 (TALL-1), or TNF-related apoptosis-inducing ligand (TRAIL).

[0075] The term "engineered Autologous Cell Therapy," which can be abbreviated as "eACT™," also known as adoptive cell transfer, is a process by which a patient's own immune cells, e.g., T cells and/or NK cells, are collected and subsequently genetically altered to recognize and target one or more antigens expressed on the cell surface of one or more specific tumor cells or malignancies. Immune cells, e.g., T cells and/or NK cells, can be engineered to express, for example, chimeric antigen receptors (CAR) or T cell receptor (TCR). CAR positive (+) immune cells, e.g. T cells or immune cells, are engineered to express an extracellular single chain variable fragment (scFv) with specificity for a particular tumor antigen linked to an intracellular signaling part comprising a costimulatory domain and an activating domain. The costimulatory domain can be derived from, e.g., CD28, and the activating domain can be derived from, e.g., CD3-zeta (figure 1). In certain embodiments, the CAR is designed to have two, three, four, or more costimulatory domains. The CAR scFv can be designed to target, for example, CD 19, which is a transmembrane protein expressed by cells in the B cell lineage, including all normal B cells and B cell malignances, including but not limited to NHL, CLL, and non-T cell ALL. Example CAR+ T cell therapies and constructs are described in U.S. Patent Publication Nos. 2013/0287748, 2014/0227237, 2014/0099309, and 2014/0050708, and these references are incorporated by reference in their entirety.

[0076] The term "Fas" as used herein refers to the Fas receptor protein or a portion thereof. The Fas receptor (also known as Fas, FasR, apoptosis antigen 1, APO-1, APT, CD95, and TNFRSF6) is a cell surface receptor protein for Fas ligand. The canonical amino acid sequence for the human Fas receptor is shown in Table 1 (UniProt P25445; SEQ ID NO: 30). Binding of Fas ligand to Fas receptor on a cell leads to apoptosis of the cell. Some aspects of the present disclosure are directed to immune cells modified to express one or more chimeric polypeptide comprising the extracellular ligand-binding domain of the Fas receptor (Fas ECD) (e.g., SEQ ID NO: 31) linked to a heterologous intracellular domain, e.g., a CD27 intracellular domain (e.g., SEQ ID NO: 22), a 4-1BB intracellular domain (e.g., SEQ ID NO: 23), or an 0X40 intracellular domain (e.g., SEQ ID NOs: 24-26). Non-limiting examples of chimeric Fas proteins that can be used in the compositions and methods disclosed herein can be found in Table 1. In some aspects, the Fas is a Fas dominant negative, or "FasDN." As used herein, "FasDN" refers to a Fas receptor that is modified to comprise a truncated Fas intracellular domain. An example of a FasDN sequence that can be used in the compositions and methods disclosed herein is presented in Table 1 (SEQ ID NO: 27). In some aspects, the chimeric Fas polypeptide further comprises a signal peptide. Any signal peptide capable of facilitating expression of the chimeric Fas polypeptide can be used in the compositions and methods disclosed herein. In some aspects, the signal peptide comprises the amino acid sequences set forth in SEQ ID NO: 28. In some aspects, the signal peptide comprises the amino acid sequences set forth in SEQ ID NO: 29.

Table 1. Example Fas Constructs [0077] An "immune response" is as understood in the art, and generally refers to a biological response within a vertebrate against foreign agents or abnormal, e.g., cancerous cells, which response protects the organism against these agents and diseases caused by them. An immune response is mediated by the action of one or more cells of the immune system (for example, a T lymphocyte, B lymphocyte, natural killer (NK) cell, macrophage, eosinophil, mast cell, dendritic cell or neutrophil) and soluble macromolecules produced by any of these cells or the liver (including antibodies, cytokines, and complement) that results in selective targeting, binding to, damage to, destruction of, and/or elimination from the vertebrate's body of invading pathogens, cells or tissues infected with pathogens, cancerous or other abnormal cells, or, in cases of autoimmunity or pathological inflammation, normal human cells or tissues. An immune reaction includes, e.g., activation or inhibition of a T cell, e.g., an effector T cell, a Th cell, a CD4⁺ cell, a CD8⁺ T cell, or a Treg cell, or activation or inhibition of any other cell of the immune system, e.g., NK cell. In some aspects, an immune response refers to NK cell-mediated killing of a foreign cell, e.g., an allogeneic T cell therapy.

[0078] "Immunotherapy" refers to the treatment of a subject afflicted with, or at risk of contracting or suffering a recurrence of, a disease by a method comprising inducing, enhancing, suppressing or otherwise modifying the immune system or an immune response.

[0079] As used herein, the term "inactivating" or "inactivation," e.g., in reference to a gene or protein, refers to a measure that can induce the reduced expression of the protein. In some aspects, inactivation can be achieved by a deletion or mutation of all or a part of the coding region of a gene or all or part of a non-coding region of a gene that results in decreased expression of the gene or the protein encoded by the gene. In some aspects, inactivation is achieved by deletion of the entire coding region of a gene. In some aspects, inactivation is achieved by partial deletion of a coding region of a gene. In some aspects, inactivation is achieved by deletion of one or more regulatory elements that facilitate expression of the gene. In some aspects, inactivation is achieved by a mutation in one or more regulatory elements that results in decreased expression or loss of expression of the gene. In some aspects, inactivation is achieved by mutation of one or more nucleic acid that results in the expression of a non-functional protein. In some aspects, inactivation is achieved by a missense mutation that results in the expression of a non-functional protein. In some aspects, inactivation is achieved by interference of the transcription or translation of a gene that results in the reduced expression of the protein. In some aspects, decreased expression is relative to the expression of the target gene in the cell prior to modification (e.g., deletion or mutation). In some aspects, the expression of the gene is measured prior to modification, then the cell is modified, and then the expression of the gene is measured following modification.

[0080] As used herein, the term "introducing" or "introduction" refers to expressing a heterologous polynucleotide and/or polypeptide in a cell. In some aspects, introduction is achieved by transfecting the cell with a polynucleotide of interest. In some aspects, introduction is achieved by genetically modifying the cell to express a heterologous sequence, e.g., using a gene editing tool including, but not limited to, CRISPR/Cas, CRISPR/Cas9, CRISPR/Cas 12, CRISPR/Cas 12a, CRISPR/Cpfl, zinc finger, TALEN, Closver-Cas or a variant thereof. In some aspects, introduction is achieved by contacting the cell with an mRNA encoding a polypeptide of interest, such that the mRNA enters into the cell or the nucleus of the cell. In some aspects, introduction comprises transfecting or transducing a cell with a polynucleotide encododing a popypeptide.

[0081] As used herein, the term "iPS cell" or "iPSC" refers to a cell that has been dedifferentiated (or reprogrammed) into a more naive, e.g., pluripotent, state. Various methods of de-differentiating a cell are known, including, but not limited to, overexpressing Oct3/4, Sox2, Klf4, and c-Myc (the '"Yamanaka factors") in the cell (see, e.g., Takahashi and Yamanaka, Cell 126.663-^r16 (2006)). In some aspects, the iPS cell is a pluripotent cell, e.g, capable of differentiation into a limited number of cell types. In some aspects, the iPS cell is a totipotent cell, e.g, capable of differentiation into any cell type. In some aspects, an iPS cell can be re- differentiated into a specific type of cell, e.g., an immune cell.

[0082] The term "lymphocyte" as used herein includes natural killer (NK) cells, T cells, or B cells. NK cells are a type of cytotoxic (cell toxic) lymphocyte that represent a major component of the inherent immune system. NK cells reject tumors and cells infected by viruses by inducing apoptosis or programmed cell death in the target cell. They were termed "natural killers" because NK cells do not require activation in order to kill a target cell. T-cells play a major role in cell-mediated-immunity. T-cell receptors (TCR) expressed on the surface of T cells differentiate T cells from other lymphocyte types. The thymus, a specialized organ of the immune system, is primarily responsible for T cell maturation. There are six types of T-cells, namely: Helper T-cells (e.g. CD4+ cells); Cytotoxic T-cells (also known as TC, cytotoxic T lymphocyte, CTL, T-killer cell, cytolytic T cell, CD8+ T-cells or killer T cell); Memory T-cells ((i) stem memory TSCM cells, like naive cells, are CD45RO-, CCR7+, CD45RA+, CD62L+ (L-selectin), CD27+, CD28+ and IL-7Ra+, but they also express large amounts of CD95, IL-2R.p, CXCR3, and LFA-1, and show numerous functional attributes distinctive of memory cells); (ii) central memory TCM cells express L-selectin and the CCR7, they secrete IL-2, but not IFNy or IL-4, and (iii) effector memory TEM cells, however, do not express L-selectin or CCR7 but produce effector cytokines like IFNy and IL-4); Regulatory T-cells (Tregs, suppressor T cells, or CD4+CD25+ regulatory T cells); Natural Killer T-cells (NKT); and Gamma Delta T-cells.

[0083] B-cells play a principal role in humoral immunity (with antibody involvement). A B cell makes antibodies and antigens and performs the role of antigen-presenting cells (APCs) and turns into memory B-cells after activation by antigen interaction. In mammals, immature B-cells are formed in the bone marrow, where its name is derived from.

[0084] As used herein, an "MHC class I molecule" refers to a protein product of a wildtype or variant HLA class I gene encoding an MHC class I molecule. Accordingly, "HLA class I molecule" and "MHC class I molecule" can be used interchangeably. The MHC Class I molecule comprises two protein chains: the alpha chain and the p2-microglobulin (P2m) chain. Human P2m is encoded by the B2M gene. The amino acid sequence of P2m is set forth in SEQ ID NO: 21 (Table 2). The alpha chain of the MHC Class I molecule is encoded by the HLA gene complex. The HLA complex is located within the 6p21.3 region on the short arm of human chromosome 6 and contains more than 220 genes of diverse function. The HLA gene are highly variant, with over 20,000 HLA alleles and related alleles, including over 15,000 HLA Class I alleles, known in the art, encoding thousands of HLA proteins, including over 10,000 HLA Class I proteins. There are at least three genes in the HLA complex that encode an MHC Class I alpha chain protein: HLA-A, HLA-B, and HLA-C. In addition, HLA-E, HLA-F, and HLA-G encode proteins that associate with the MHC Class I molecule.

Table 2. Amino Acid Sequence of Human P2m [0085] As used herein, an "MHC class II molecule" refers to a protein product of a wildtype or variant HLA class II gene encoding an MHC class II molecule. Accordingly, "MHC class II molecule" can be used interchangeably with an "HLA class II molecule." A typical MHC Class II molecule comprises two protein chains: an alpha chain and a beta chain. In general, naturally occurring alpha chains and beta chains each comprise a transmembrane domain, which anchors the alpha/beta chain to the cell surface, and an extracellular domain, which carries the antigen and interacts with a TCR and/or CD4 expressed on a T cell. Both the MHC Class II alpha and beta chains are encoded by the HLA gene complex. The HLA complex is located within the 6p21.3 region on the short arm of human chromosome 6 and contains more than 220 genes of diverse function. The HLA gene complex is highly variant, with over 20,000 HLA alleles and related alleles, including over 250 MHC class II alpha chain alleles and 5,000 MHC class II beta chain alleles, known in the art, encoding thousands of MHC class II proteins. Three loci in the HLA complex encode MHC Class II proteins: HLA-DP, HLA-DQ, and HLA-DR. HLA- DO and HLA-DM encode proteins that associate with the MHC class II molecule and support its configuration and function.

[0086] As used herein, "pharmaceutically acceptable carrier" includes any and all aqueous solvents (e.g., water, alcoholic/aqueous solutions, saline solutions, parenteral vehicles, such as sodium chloride, Ringer's dextrose, etc.), non-aqueous solvents (e.g., propylene glycol, polyethylene glycol, vegetable oil, and injectable organic esters, such as ethyloleate), dispersion media, coatings, surfactants, antioxidants, preservatives (e.g., antibacterial or antifungal agents, anti-oxidants, chelating agents, and inert gases), isotonic agents, absorption delaying agents, salts, drugs, drug stabilizers, gels, binders, excipients, disintegration agents, lubricants, sweetening agents, flavoring agents, dyes, fluid and nutrient replenishers, such like materials and combinations thereof, as would be known to one of ordinary skill in the art. The pH and exact concentration of the various components in a pharmaceutical composition are adjusted according to well-known parameters.

[0087] "Potentiating an endogenous immune response" means increasing the effectiveness or potency of an existing immune response in a subject. This increase in effectiveness and potency can be achieved, for example, by overcoming mechanisms that suppress the endogenous host immune response or by stimulating mechanisms that enhance the endogenous host immune response. [0088] The term "recombinant" or "modified" cell, as used herein, is intended to refer to a cell, e.g., an immune cell, that comprises a nucleic acid that is not naturally present in the cell, and can be a cell into which a recombinant expression vector has been introduced. It should be understood that such terms are intended to refer not only to the particular subject cell but also to the progeny of such a cell. Though certain modifications can occur in succeeding generations due to either mutation or environmental influences, such progeny are still included within the scope of the term "recombinant" or "modified" as used herein.

[0089] As used herein, the terms "reduced expression" and "increased expression" refer to the expression of a particular gene or protein in a cell relative to a control, e.g., the expression of a particular gene in a modified cell as compared to the expression of the gene in a wild-type (unmodified) cell. The relative expression can be based on mRNA levels and/or protein levels. Any means of measuring the level of mRNA and/or protein can be used to determine whether a gene or protein has reduced or increased expression, including but not limited to immunohistochemistry and PCR-based techniques.

[0090] In some aspects, a cell that has "reduced expression" of a particular gene or protein has an expression level that is at less than about 99%, less than about 98%, less than about 97%, less than about 96%, less than about 95%, less than about 90%, less than about 85%, less than about 80%, less than about 75%, less than about 70%, less than about 65%, less than about 60%, less than about 55%, less than about 50%, less than about 45%, less than about 40%, less than about 35%, less than about 30%, less than about 25%, less than about 20%, less than about 15%, less than about 10%, or less than about 5% that of the expression of the gene or protein in an unmodified cell, e.g., wildtype cell of the same cell type. In some aspects, the unmodified cell, e.g., wild-type cell of the same cell type, expresses the particular gene or protein, and the modified cell has no detectable level of expression of the protein or gene, e.g., a "knock-out" of the gene or protein. As such, the term "knock-out" refers to the complete ablation of expression of a particular gene or protein, such that there is no detectable level of expression of the gene or protein in the cell.

[0091] In some aspects, a cell that has "increased expression" or “overexpression” of a particular gene or protein has an expression level that is more than about 105%, more than about 110%, more than about 115%, more than about 120%, more than about 125%, more than about 130%, more than about 140%, more than about 150%, more than about 160%, more than about 170%, more than about 180%, more than about 190% more than about 200%, more than about 225%, more than about 250%, more than about 275%, more than about 300%, more than about 350%, more than about 400%, more than about 450%, more than about 500%, more than about 600%, more than about 700%, more than about 800%, more than about 900%, or more than about 1000% that of the expression of the gene or protein in an unmodified cell, e.g., a wild-type cell of the same cell type or population of cells or the same cell or population of cells prior to the modification, wherein 100% expression corresponds to that observed in case of the wild-type cell. In some aspects, a cell that has "increased expression" or “overexpression” of a particular gene or protein has an expression level that at least about 5% higher, at least about 10% higher, at least about 15% higher, at least about 20% higher, at least about 25% higher, at least about 30% higher, at least about 35% higher, at least about 40% higher, at least about 45% higher, at least about 50% higher, at least about 55% higher, at least about 60% higher, at least about 65% higher, at least about 70% higher, at least about 75% higher, at least about 80% higher, at least about 85% higher, at least about 90% higher, at least about 95% higher, at least about 100% higher, at least about 110% higher, at least about 120% higher, at least about 130% higher, at least about 140% higher, at least about 150% higher, at least about 160% higher, at least about 170% higher, at least about 180% higher, at least about 190% higher, at least about 200% higher, at least about 250% higher, at least about 300% higher, at least about 350% higher, at least about 400% higher, at least about 450% higher, at least about 500% higher, at least about 600% higher, at least about 700% higher, at least about 800% higher, at least about 900% higher, or at least about 1000% higher than the expression of the gene or protein in an unmodified cell, e.g., a wild-type cell of the same cell type or population of cells or the same cell or population of cells prior to the modification. In some aspects, the unmodified cell, e.g., wild-type cell of the same cell type or the same cell or population of cells prior to the modification, has no expression of the particular gene or protein, and the increased expression is any expression of the gene or protein.

[0092] Increased expression of a particular gene or protein can be achieved by any method. In some aspects, the expression of a gene or polypeptide is increased by introducing into the cell, a molecule, signal, element or modification that results in increased expression of a gene or polypeptide in the cell. In some aspects, the expression of a gene or polypeptide is increased by transfecting the cell with a nucleic acid molecule encoding a protein. In some aspects, a nucleic acid of interest is introduced into the cell via eletroporation. In some aspects, the nucleic acid is a vector. In some aspects, the nucleic acid comprises an mRNA. In some aspects, the expression of a gene or protein is increased by modifying an endogenous regulatory element or inserting a heterolgous regulatory element into an endogenous gene thereby to increase expression of the endogenous gene encoding a polypeptide of interest. In some aspects, the expression of a gene or protein is increased by knocking-in a heterolgous coding region that encodes the polypeptide of interest. In some aspects, modification of an endogenous sequence is achieved using a gene editing tool, such as CRISPR.

[0093] As used herein, the terms "subject" and "patient" are used interchangeably and refer to either a human or a non-human, such as primates, mammals, and vertebrates. In particular aspects, the subject is a human.

[0094] The term "chimeric antigen receptor" or "CAR," as used herein, refers to a recombinant fusion protein that has an antigen-specific extracellular domain coupled to an intracellular domain that directs the cell to perform a specialized function upon binding of an antigen to the extracellular domain. In some aspects, a chimeric antigen receptor disclosed herein comprises a chimeric polypeptide of the present disclosure.

[0095] The term "T cell receptor" (TCR), as used herein, refers to a heteromeric cellsurface receptor capable of specifically interacting with a target antigen. As used herein, "TCR" includes but is not limited to naturally occurring and non-naturally occurring TCRs; full-length TCRs and antigen binding portions thereof; chimeric TCRs; TCR fusion constructs; and synthetic TCRs. In human, TCRs are expressed on the surface of T cells, and they are responsible for T cell recognition and targeting of antigen presenting cells. Antigen presenting cells (APCs) display fragments of foreign proteins (antigens) complexed with the major histocompatibility complex (MHC; also referred to herein as complexed with an HLA molecule, e.g., an HLA class 1 molecule). A TCR recognizes and binds to the antigen-HLA complex and recruits CD3 (expressed by T cells), activating the TCR. The activated TCR initiates downstream signaling and an immune response, including the destruction of the antigen-presenting cell.

[0096] An "exogenous TCR" or an "eTCR," as used herein, refers to a TCR that is heterologous to the cell which expresses the TCR. As used herein, the term "heterologous" refers to something that is not native to or naturally found in, e.g., the paticular cell. [0097] In general, a TCR can comprise two chains, (i) an alpha chain and a beta chain (alpha-beta TCR) for alpha-beta T cells, or (ii) a gamma chain and a delta chain (gammadelta TCR) for gamma-delta T cells, interconnected by disulfide bonds. Each chain comprises a variable domain (alpha chain variable domain, beta chain variable domain, gamma chain variable domain, and delta chain variable domain) and a constant region (alpha chain constant region, beta chain constant region, gamma chain constant region, and delta chain constant region). The variable domain is located distal to the cell membrane, and the variable domain interacts with an antigen. The constant region is located proximal to the cell membrane. A TCR can further comprises a transmembrane region and a short cytoplasmic tail. As used herein, the term "constant region" encompasses the transmembrane region and the cytoplasmic tail, when present, as well as the traditional "constant region."

[0098] The variable domains can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDRs), interspersed with regions that are more conserved, termed framework regions (FR). Each alpha chain variable domain and beta chain variable domain comprises three CDRs and four FRs: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. Each variable domain contains a binding domain that interacts with an antigen. Though all three CDRs on each chain are involved in antigen binding, CDR3 is believed to be the primary antigen binding region. CDR1 is also interacts with the antigen, while CD2 is believed to primarily recognize the ELLA complex. In one embodiment, the gamma chain variable domain include V gamma 1, V gamma 2, V gamma 3, V gamma 4, V gamma 5, V gamma 6, V gamma 7, V gamma 8 and V gamma 9, and examples of delta chain variable domain include V delta 1, V delta 2, V delta 3, V delta 4, V delta 5, V delta 6, V delta 7, V delta 8 and V delta 9. While the combination of specific gamma chain variable domain and delta chain variable domain in the TCR is not limited, for example, eTCR is any of V gamma 3-V delta 1 TCR (g3dlTCR), V gamma 4-V delta 1 TCR (g4dlTCR), V gamma 9-V delta 1 TCR (g9dlTCR) and V gamma 9-V delta 2 TCR (g9d2TCR).

[0099] Where not expressly stated, and unless the context indicates otherwise, the term "TCR" also includes an antigen-binding fragment or an antigen-binding portion of any TCR disclosed herein, and includes a monovalent and a divalent fragment or portion, and a single chain TCR. The term "TCR" is not limited to naturally occurring TCRs bound to the surface of a T cell. As used herein, the term "TCR" further refers to a TCR described herein that is expressed on the surface of a cell other than a T cell (e.g., a cell that naturally expresses or that is modified to express CD3, as described herein), or a TCR described herein that is free from a cell membrane (e.g., an isolated TCR or a soluble TCR).

[0100] A "TCR fragment," "antigen binding molecule," or "portion of a TCR" refers to any portion of a TCR less than the whole. An antigen binding molecule can include the antigenic complementarity determining regions (CDRs).

[0101] A "suicide gene" refers to a gene that causes a cell to kill itself. In some embodiments, the suicide gene causes a cell to kill itself through apoptosis. Non-limiting examples of suicide genes include viral thymidine kinase, cytosine deaminases, intracellular antibody against antioxidative enzymes (AOEs), bacterial nitroreductase, caspase and DNase.

[0102] In one embodiment, the suicide gene is viral thymidine kinase (TK). Thymidine kinase is an ATP -thymidine 5 '-phosphotransferase that converts deoxythymidine intodeoxythymidien 5'-monophosphate, which is further phosphorylated to deoxythymidine diphosphate and thereafter to deoxythymidine triphosphate by viral thymidine kinase and nucleoside diphosphate kinase respectively. Deoxythymidine triphosphate is incorporated into the synthesized DNA molecule by DNA polymerase. Some dNTP analogs, such as Ganciclovir (GCV), a synthetic analogue of 2'-deoxy- guanosine, have the ability to terminate the DNA synthesis upon their incorporation into synthesized DNA. Termination of synthesis triggers the apoptotic signaling cascades. While GCV is not recognized by human thymidine kinase, it is recognized as a substrate for some viral thymidine kinase, such as Herpes Simplex Virus- 1 thymidine kinase (HSV-TK). As a result, a human cell expressing HSV-TK converts GCV into GCV phosphate, which is further phosphorylated and incorporated into the synthesized DNA, leading to the termination of synthesis and apoptosis. While the variant of HSV-TK is not limited, HSV-TK is, for example, TK007 (see Preuss et al., Hum Gene Ther. 2010 Aug; 21(8): 929-41).

[0103] In some embodiments, the suicide gene is cytosine deaminase. Cytosine deaminase hydrolyze cytosine to uracile with release of ammonia. In physiological conditions, the modified site is recognized by endonucleases, then the phophodiester bond in the DNA is broken, initiating repair by incorporation of a new cytosine. However, cytosine deaminase can also converts 5-fluorocytosine into 5 -fluorouracil (5-FU). Therefore, upon provision of non-toxic prodrug 5-FC, cytosine deaminase converts it into highly toxic 5-FU (a suicide inhibitor of thymidylate synthetase), leading to the inhibition of cell growth and apoptosis.

[0104] The term "therapeutic benefit" or "therapeutically effective," as used herein, refers to anything that promotes or enhances the well-being of the subject with respect to the medical treatment of this condition. This includes, but is not limited to, a reduction in the frequency or severity of the signs or symptoms of a disease.

[0105] As used herein, the term "treating" or "treatment" of a disease or condition refers to executing a protocol, which may include administering one or more therapies to a patient, in an effort to alleviate signs or symptoms of the disease. In some aspects, a treatment decreases the rate of disease progression, ameliorates or palliates the disease state, and/or facilitates remission or improved prognosis. Alleviation can occur prior to signs or symptoms of the disease or condition appearing, as well as after their appearance. Thus, in some aspects, "treating" or "treatment" includes "preventing" or "prevention" of a disease or an undesirable condition. However, "treating" or "treatment" does not require complete alleviation of all signs and/or symptoms, does not require a cure, and specifically includes protocols that have only a marginal effect on the patient.

[0106] In various aspects, a subject in need thereof may be treated for a disease or for alleviating symptoms associated with a disease (e.g., a cancer) using a population of modified cells described herein. In some aspects, the modified cells are immune cells or iPS cells that are transduced with a CAR or a TCR and are further modified to exhibit reduced expression of endogenous MHC class I and MHC class II HLAs genes in combination with expression or increased expression of certain heterologous genes in the cell, such that the modified cells are less susceptible to killing by the subject’s immune system when such cells are administered to the subject .

[0107] As used herein, the terms "ug" and "uM" are used interchangeably with "pg" and "pM," respectively.

[0108] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure is related. For example, the Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd ed., 2002, CRC Press; The Dictionary of Cell and Molecular Biology, 3rd ed., 1999, Academic Press; and the Oxford Dictionary of Biochemistry and Molecular Biology, Revised, 2000, Oxford University Press, provide one of skill with a general dictionary of many of the terms used in this disclosure.

[0109] Units, prefixes, and symbols are denoted in their Systeme International de Unites (SI) accepted form. Numeric ranges are inclusive of the numbers defining the range. As described herein, any concentration range, percentage range, ratio range or integer range is to be understood to include the value of any integer within the recited range and, when appropriate, fractions thereof (such as one-tenth and one-hundredth of an integer), unless otherwise indicated.

[0110] Abbreviations used herein are defined throughout the present disclosure. Various aspects of the disclosure are described in further detail in the following subsections.

[OHl] Various aspects described herein are described in further detail in the following subsections.

II. Compositions of the Disclosure

[0112] Some aspects of the present disclosure are directed to modified cells, comprising: (i) a chimeric antigen receptor (CAR) and/or an exogenous T cell receptor (eTCR); (ii) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; and (iv) a polypeptide comprising a dominant-negative Fas (Fas-DN), a Fas-CD27 chimeric polypeptide (Fas-CD27), a Fas-4-lBB chimeric polypeptide (Fas-BB), a Fas-OX40 chimeric polypeptide (Fas-OX40), or any combination thereof.

[0113] Other aspects of the present disclosure are directed to a modified cell comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN); (iv) increased expression of HLA-E relative to a wild-type cell of the same cell type; (v) a nucleic acid encoding a suicide gene; or (vi) any combination thereof.

[0114] Other aspects of the present disclosure are directed to a modified cell comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); and (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene, or any combination thereof.

[0115] Other aspects of the present disclosure are directed to a modified cell comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); and

(iii) inactivated endogenous gene encoding CD58.

[0116] Other aspects of the present disclosure are directed to a modified cell comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; and (iv) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof.

[0117] Other aspects of the present disclosure are directed to a modified cell comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; and (iv) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide and a nucleic acid encoding a suicide gene.

[0118] Other aspects of the present disclosure are directed to a dominant-negative Fas (Fas-DN), a Fas-CD27 chimeric polypeptide (Fas-CD27), a Fas-4-lBB chimeric polypeptide (Fas-BB), or a Fas-OX40 chimeric polypeptide (Fas-OX40).

[0119] Other aspects of the present disclosure are directed to a guide RNA capable of hybridizing to a human CD58 gene.

II. A. Modified Cells

[0120] Some aspects of the present disclosure are directed to modified cells, comprising: (i) a chimeric antigen receptor (CAR) and/or an exogenous T cell receptor (eTCR); (ii) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; and

(iv) a polypeptide comprising a dominant-negative Fas (Fas-DN), a Fas-CD27 chimeric polypeptide (Fas-CD27), a Fas-4-lBB chimeric polypeptide (Fas-BB), a Fas-OX40 chimeric polypeptide (Fas-OX40), or any combination thereof. [0121] In some aspects, the cells disclosed herein have has increased persistence in vivo relative to a wild-type cell of the same cell type. In some aspects, the cell disclosed herein are less immunogenic when administered to a human subject. In some aspects, the human subject has less of an immune response against the cells following administration. In some aspects, the human subject has reduced NK cell killing of the cells following administration. In some aspects, NK cell mediated killing of the cells following administration is reduced by at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% relative to a wild-type cell of the same cell type administered to a human subject.

[0122] In some aspects, the MHC-I human leukocyte antigens are HLA-A, HLA-B and HLA-C. In some aspects, the expression of the MHC-I human leukocyte antigen is less than about 99%, less than about 98%, less than about 97%, less than about 96%, less than about 95%, less than about 90%, less than about 85%, less than about 80%, less than about 75%, less than about 70%, less than about 65%, less than about 60%, less than about 55%, less than about 50%, less than about 45%, less than about 40%, less than about 35%, less than about 30%, less than about 25%, less than about 20%, less than about 15%, less than about 10%, or less than about 5% the MHC-I human leukocyte antigen expression in a wild-type cell of the same cell type. In some aspects, the cell has no detectable expression of the MHC-I human leukocyte antigen.

[0123] In some aspects, the reduced expression of the endogenous MHC-I human leukocyte antigens results from a mutation in, or a deletion of one or more endogenous genes encoding beta-2-microglobulin (B2M). In some aspects, the expression of the B2M is less than about 99%, less than about 98%, less than about 97%, less than about 96%, less than about 95%, less than about 90%, less than about 85%, less than about 80%, less than about 75%, less than about 70%, less than about 65%, less than about 60%, less than about 55%, less than about 50%, less than about 45%, less than about 40%, less than about 35%, less than about 30%, less than about 25%, less than about 20%, less than about 15%, less than about 10%, or less than about 5% the B2M expression in a wild-type cell of the same cell type. In some aspects, the cell has no detectable expression of the B2M. [0124] In some aspects, the MHC-II human leukocyte antigens are HLA-DP, HLA-DQ and HLA-DR. In some aspects, the expression of the MHC-II human leukocyte antigen is less than about 99%, less than about 98%, less than about 97%, less than about 96%, less than about 95%, less than about 90%, less than about 85%, less than about 80%, less than about 75%, less than about 70%, less than about 65%, less than about 60%, less than about 55%, less than about 50%, less than about 45%, less than about 40%, less than about 35%, less than about 30%, less than about 25%, less than about 20%, less than about 15%, less than about 10%, or less than about 5% the MHC-II human leukocyte antigen expression in a wild-type cell of the same cell type. In some aspects, the cell has no detectable expression of the MHC-II human leukocyte antigen.

[0125] In some aspects, the reduced expression of the endogenous endogenous MHC-II human leukocyte antigens results from a mutation in, or a deletion of the one or more endogenous genes encoding class II major histocompatibility complex transactivator (CIITA). In some aspects, the expression of the CIITA is less than about 99%, less than about 98%, less than about 97%, less than about 96%, less than about 95%, less than about 90%, less than about 85%, less than about 80%, less than about 75%, less than about 70%, less than about 65%, less than about 60%, less than about 55%, less than about 50%, less than about 45%, less than about 40%, less than about 35%, less than about 30%, less than about 25%, less than about 20%, less than about 15%, less than about 10%, or less than about 5% the CIITA expression in a wild-type cell of the same cell type. In some aspects, the cell has no detectable expression of the CIITA.

[0126] In some aspects, the cell further comprises a reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type. In some aspects, the reduced expression of the CD58 results from a mutation in, or a deletion of, one or more endogenous genes encoding CD58. In some aspects, the expression of CD58 is less than about 99%, less than about 98%, less than about 97%, less than about 96%, less than about 95%, less than about 90%, less than about 85%, less than about 80%, less than about 75%, less than about 70%, less than about 65%, less than about 60%, less than about 55%, less than about 50%, less than about 45%, less than about 40%, less than about 35%, less than about 30%, less than about 25%, less than about 20%, less than about 15%, less than about 10%, or less than about 5% the CD58 expression a wild-type cell of the same cell type. In some aspects, the cell has no detectable expression of functional CD58. II.A.l. Expression of Fas Constructs

[0127] In some aspects, the cell further comprises a nucleic acid encoding a Fas-DN, e.g., a Fas-DN disclosed herein. Accordingly, some aspects of the present disclosure are directed to a modified cell, e.g., immune cell, comprising: (i) a CAR or an eTCR; (ii) reduced expression of endogenous MHC-I human leukocyte antigens relative to a wildtype cell of the same cell type; (iii) reduced expression of endogenous MHC-II human leukocyte antigens relative to a wild-type cell of the same cell type; and (iv) a Fas-DN. In some aspects the modified cell, e.g., immune cell, comprises: (i) a CAR or an eTCR; (ii) reduced expression of endogenous B2M; (iii) reduced expression of endogenous CIITA; and (iv) a Fas-DN. In some aspects the modified cell, e.g., immune cell, comprises: (i) a CAR or an eTCR; (ii) reduced expression of endogenous B2M; (iii) reduced expression of endogenous CIITA; (iv) reduced expression of endogenous CD58 relative to a wildtype cell of the same cell type; and (v) a Fas-DN.

[0128] In some aspects, the Fas-DN is encoded by a nucleotide sequence having at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identity to the nucleotide sequence set forth in SEQ ID NO: 35, 36, or 40. In some aspects, the Fas-DN is encoded by a nucleotide set forth in SEQ ID NO: 35, 36, or 40.

Table 3: Coding Sequences

[0129] In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type; and (iv) a Fas-DN. In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of emdogenous CIITA relative to a wild-type cell of the same cell type; and (iv) a Fas-DN. In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type; and (v) a Fas-DN.

[0130] In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type; and (iv) a Fas-DN. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; and (iv) a Fas-DN. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type; and (v) a Fas-DN.

[0131] In some aspects, the cell further comprises a Fas-CD27, e.g., a Fas-CD27 disclosed herein. Accordingly, some aspects of the present disclosure are directed to a modified cell, e.g., immune cell, comprising: (i) a CAR or an eTCR; (ii) reduced expression of endogenous MHC-I human leukocyte antigens relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II human leukocyte antigens relative to a wild-type cell of the same cell type; and (iv) a Fas-CD27. In some aspects the modified cell, e.g., immune cell, comprises: (i) a CAR or an eTCR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; and (iv) a Fas-CD27. In some aspects the modified cell, e.g., immune cell, comprises: (i) a CAR or an eTCR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type; and (v) a Fas-CD27.

[0132] In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type; and (iv) a Fas-CD27. In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of CIITA; and (iv) a nucleic acid encoding Fas-CD27. In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of B2M; (iii) reduced expression of CIITA; (iv) reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type; and (v) a nucleic acid encoding Fas-CD27.

[0133] In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type; and (iv) a Fas-CD27. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; and (iv) a Fas-CD27. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type; and (v) a Fas-CD27.

[0134] In some aspects, the cell further comprises a Fas-4-lBB, e.g., a Fas-4-lBB disclosed herein. Accordingly, some aspects of the present disclosure are directed to a modified cell, e.g., immune cell, comprising: (i) a CAR or an eTCR; (ii) reduced expression of endogenous MHC-I human leukocyte antigens relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II human leukocyte antigens relative to a wild-type cell of the same cell type; and (iv) a Fas-4-lBB. In some aspects the modified cell, e.g., immune cell, comprises: (i) a CAR or an eTCR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; and (iv) a Fas-4-lBB. In some aspects the modified cell, e.g., immune cell, comprises: (i) a CAR or an eTCR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type; and (v) a Fas-4-lBB.

[0135] In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type; and (iv) a Fas-4-lBB. In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; and (iv) a Fas-4-lBB. In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type; and (v) a Fas-4-lBB.

[0136] In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type; and (iv) a Fas-4-lBB. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; and (iv) a Fas-4-lBB. In some aspects, the modified cells comprise: (i) an eTCR;

(ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type; and (v) a Fas-4-lBB.

[0137] In some aspects, the cell further comprises a Fas-OX40, e.g., a Fas-OX40 disclosed herein. Accordingly, some aspects of the present disclosure are directed to a modified cell, e.g., immune cell, comprising: (i) a CAR or an eTCR; (ii) reduced expression of endogenous MHC-I human leukocyte antigens relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II human leukocyte antigens relative to a wild-type cell of the same cell type; and (iv) a Fas-OX40. In some aspects the modified cell, e.g., immune cell, comprises: (i) a CAR or an eTCR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type;

(iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; and (iv) a Fas-OX40. In some aspects the modified cell, e.g., immune cell, comprises: (i) a CAR or an eTCR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type; and (v) a Fas-OX40. [0138] In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type; and (iv) a Fas-OX40. In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; and (iv) a Fas-OX40. In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type; and (v) a Fas-OX40.

[0139] In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type; and (iv) a Fas-OX40. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; and (iv) a Fas-OX40. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type; and (v) a Fas-OX40.

II.A.2. Modified Expression of Endogenous Poliovirus Receptor (PVR)

[0140] In some aspects, the modified cell further comprises reduced expression of an endogenous poliovirus receptor (PVR) relative to a wild-type cell of the same cell type. In some aspects, the reduced expression of the PVR results from a mutation in, or a deletion of, one or more endogenous genes encoding PVR. In some aspects, the expression of PVR is less than about 99%, less than about 98%, less than about 97%, less than about 96%, less than about 95%, less than about 90%, less than about 85%, less than about 80%, less than about 75%, less than about 70%, less than about 65%, less than about 60%, less than about 55%, less than about 50%, less than about 45%, less than about 40%, less than about 35%, less than about 30%, less than about 25%, less than about 20%, less than about 15%, less than about 10%, or less than about 5% the PVR expression a wild-type cell of the same cell type. In some aspects, the cell has no detectable expression of functional PVR. In some aspects, the cell has no detectable expression of PVR.

[0141] In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; and (v) a Fas-DN. In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; and (v) a Fas-DN. In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type;

(iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type; (v) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; and (vi) a Fas-DN.

[0142] In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type;

(iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; and (v) a Fas-DN. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; and (v) a Fas-DN. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type; (v) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; and (vi) a Fas-DN. [0143] In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; and (v) a Fas-CD27. In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type;

(iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; and (v) a Fas-CD27. In some aspects, the modified cells comprise: (i) a CAR;

(ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type; (v) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; and (vi) a Fas-CD27.

[0144] In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type;

(iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; and (v) a Fas-CD27. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type;

(iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; and (v) a Fas-CD27. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type; (v) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; and (vi) a Fas-CD27.

[0145] In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type;

(iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; and (v) a Fas-4-lBB. In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type;

(iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; and (v) a Fas-4-lBB. In some aspects, the modified cells comprise: (i) a CAR;

(ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type; (v) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; and (vi) a Fas-4-lBB.

[0146] In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type;

(iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; and (v) a Fas-4-lBB. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type;

(iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; and (v) a Fas-4-lBB. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenoud B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type; (v) reduced expression of endogenous PVR relative to a wildtype cell of the same cell type; and (vi) a Fas-4-lBB.

[0147] In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type;

(iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; and (v) a Fas-OX40. In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; and (v) a Fas-OX40. In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type; (v) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; and (vi) a Fas-OX40.

[0148] In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of MHC-II HLA relative to a wild-type cell of the same cell type; (iv) reduced expression of PVR; and (v) a nucleic acid encoding Fas-OX40. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of B2M; (iii) reduced expression of CIITA; (iv) reduced expression of PVR; and (v) a nucleic acid encoding Fas-OX40. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of B2M; (iii) reduced expression of CIITA; (iv) reduced expression of CD58; (v) reduced expression of PVR; and (vi) a nucleic acid encoding Fas-OX40.

II. A.3. Modified Expression ofHLA-E

[0149] In some aspects, the modified cell further comprises increased expression of HLA-E relative to a wild-type cell of the same cell type, e.g., the cell prior to modification. In some aspects, the increased expression of the HLA-E results from transfection of a nucleic acid encoding an HLA-E polypeptide. In some aspects, the increased expression of the HLA-E results from a modification to an endogenous gene encoding HLA-E polypeptide, wherein the modification increases the expression of the endogenous HLA-E polypeptide. In some aspects, the increased expression of the HLA-E results from (i) transfection of a nucleic acid encoding an HLA-E polypeptide and (ii) a modification to an endogenous gene encoding HLA-E polypeptide, wherein the modification increases the expression of the endogenous HLA-E polypeptide. In some aspects, the expression of the HLA-E is at least about 105%, at least about 110%, at least about 115%, at least about 120%, at least about 125%, at least about 130%, at least about 140%, at least about 150%, at least about 160%, at least about 170%, at least about 180%, at least about 190% at least about 200%, at least about 225%, at least about 250%, at least about 275%, at least about 300%, at least about 350%, at least about 400%, at least about 450%, at least about 500%, at least about 600%, at least about 700%, at least about 800%, at least about 900%, or at least about 1000% that of the expression of HLA-E in a wildtype cell of the same cell type. [0150] In some aspects, the modified cell further comprises HLA-E. In some aspect, the expression of the HLA-E is at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90% at least about 100% that of the expression of HLA-E in a wild-type cell of the same cell type.

[0151] In some aspects, the HLA-E is a human HLA-E polypeptide. In some aspects, the HLA-E polypeptide is a chimeric polypeptide comprising a human HLA-E polypeptide or a portion thereof. In some aspects, the HLA-E polypeptide is a chimeric polypeptide comprising a HLA-E polypeptide linked to a human B2M polypeptide. In some aspects, the HLA-E polypeptide comprises an amino acid sequence having at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 32.

[0152] In some aspects, the HLA-E is encoded by a nucleotide sequence having at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identity to the nucleotide sequence set forth in SEQ ID NO: 33, 36, or 38. In some aspects, the HLA-E is encoded by a nucleotide sequence set forth in SEQ ID NO: 33, 36, or 38.

[0153] In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type;

(iv) increased expression of HLA-E relative to a wild-type cell of the same cell type; and

(v) a Fas-DN. In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; (v) increased expression of HLA-E relative to a wild-type cell of the same cell type; and (vi) a Fas-DN. In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; (v) increased expression of HLA-E relative to a wild-type cell of the same cell type; and (vi) a Fas-DN. In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type; (v) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; (vi) increased expression of HLA-E relative to a wild-type cell of the same cell type; and (vii) a Fas-DN.

[0154] In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type;

(iv) increased expression of HLA-E relative to a wild-type cell of the same cell type; and

(v) a Fas-DN. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; (v) increased expression of HLA-E relative to a wild-type cell of the same cell type; and (vi) a Fas-DN. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; (v) increased expression of HLA-E relative to a wild-type cell of the same cell type; and (vi) a nucleic acid encoding Fas-DN. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type; (v) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; (vi) increased expression of HLA-E relative to a wild-type cell of the same cell type; and (vii) a Fas- DN.

[0155] In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type; (iv) increased expression of HLA-E relative to a wild-type cell of the same cell type; and

(v) a Fas-CD27. In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type;

(iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; (v) increased expression of HLA-E relative to a wild-type cell of the same cell type; and (vi) a Fas-CD27. In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; (v) increased expression of HLA-E relative to a wild-type cell of the same cell type; and (vi) a Fas-CD27. In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type; (v) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; (vi) increased expression of HLA-E relative to a wild-type cell of the same cell type; and (vii) a Fas-CD27.

[0156] In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type;

(iv) increased expression of HLA-E relative to a wild-type cell of the same cell type; and

(v) a Fas-CD27. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; (v) increased expression of HLA-E relative to a wild-type cell of the same cell type; and (vi) a Fas-CD27. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; (v) increased expression of HLA-E relative to a wild-type cell of the same cell type; and (vi) a Fas-CD27. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type; (v) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; (vi) increased expression of HLA-E relative to a wild-type cell of the same cell type; and (vii) a Fas-CD27.

[0157] In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type;

(iv) increased expression of HLA-E relative to a wild-type cell of the same cell type; and

(v) a Fas-4-lBB. In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type;

(iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; (v) increased expression of HLA-E relative to a wild-type cell of the same cell type; and (vi) a Fas-4-lBB. In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; (v) increased expression of HLA-E relative to a wild-type cell of the same cell type; and (vi) a Fas-4-lBB. In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type; (v) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; (vi) increased expression of HLA-E relative to a wild-type cell of the same cell type; and (vii) a nucleic acid encoding Fas-4-lBB.

[0158] In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type;

(iv) increased expression of HLA-E relative to a wild-type cell of the same cell type; and

(v) a Fas-4-lBB. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; (v) increased expression of HLA-E relative to a wild-type cell of the same cell type; and (vi) a Fas-4-lBB. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; (v) increased expression of HLA-E relative to a wild-type cell of the same cell type; and (vi) a Fas-4-lBB. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type; (v) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; (vi) increased expression of HLA-E relative to a wild-type cell of the same cell type; and (vii) a Fas-4-lBB.

[0159] In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type;

(iv) increased expression of HLA-E relative to a wild-type cell of the same cell type; and

(v) a Fas-OX40. In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; (v) increased expression of HLA-E relative to a wild-type cell of the same cell type; and (vi) a Fas-OX40. In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; (v) increased expression of HLA-E relative to a wild-type cell of the same cell type; and (vi) a Fas-OX40. In some aspects, the modified cells comprise: (i) a CAR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type; (v) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; (vi) increased expression of HLA-E relative to a wild-type cell of the same cell type; and (vii) a Fas-OX40.

[0160] In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type;

(iv) increased expression of HLA-E relative to a wild-type cell of the same cell type; and

(v) a Fas-OX40. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous MHC-I HLA relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II HLA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; (v) increased expression of HLA-E relative to a wild-type cell of the same cell type; and (vi) a Fas-OX40. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; (v) increased expression of HLA-E relative to a wild-type cell of the same cell type; and (vi) a Fas-OX40. In some aspects, the modified cells comprise: (i) an eTCR; (ii) reduced expression of endogenous B2M relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous CIITA relative to a wild-type cell of the same cell type; (iv) reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type; (v) reduced expression of endogenous PVR relative to a wild-type cell of the same cell type; (vi) increased expression of HLA-E relative to a wild-type cell of the same cell type; and (vii) a Fas-OX40.

II. A.4. Additional Polypeptides

[0161] In some aspects, the modified cell further overexpresses one or more additional endogenous polypeptides. In some aspects, the modified cell further expresses one or more additional heterologous polypeptides. In some aspects, the modified cell comprises a human interleukin 15 (IL15) polypeptide. In some aspects, the modified cell overexpresses an endogenous IL 15 polypeptide. In some aspects, the modified cell expresses a heterologous IL15 polypeptide. In some aspects, the IL15 polypeptide is a membrane-bound IL 15 polypeptide. In some aspects, the IL 15 polypeptide is a membrane bound IL15/IL15Ralpha fusion polypeptide (mIL15/Ra). In some aspects, the IL 15 polypeptide comprises an IL15 sushi domain/IL15Ra fusion polypeptide (sushil5). In some aspects, the IL 15 polypeptide comprises a membrane bound-IL15/IL15Ra-LSP fusion (mIL15/Ra-LSP). In some aspects, the IL15 polypeptide comprises a short IL15 polypeptide (sIL15). In some aspects, the IL 15 polypeptide comprises a soluble IL 15 polypeptide.

[0162] In some aspects, the modified cell comprises a human chemokine (C-C motif) ligand 19 (CCL19) polypeptide. In some aspects, the modified cell overexpresses an endogenous CCL19 polypeptide. In some aspects, the modified cell expresses a heterologous CCL19 polypeptide.

[0163] In some aspects, the modified cell comprises an HSVTK polypeptide. In some aspects, the HSVTK polypeptide is encoded by a nucleotide sequence having at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identity to the nucleotide sequence set forth in SEQ ID NO: 34, 36, or 39. In some aspects, the HSVTK polypeptide is encoded by a nucleotide sequence set forth in SEQ ID NO: 34, 36, or 39.

II.A.5. Cells

[0164] Any cell type can be used in the compositions and methods disclosed herein. In some aspects, the cell is an immune cell. In some aspects, the cell is an induced pluripotent stem cell (iPSC). In some aspects, the cell is an embryonic stem cell (ESC). In some aspects, the cell is an immune cell selected from a T cell, an NK cell, an NKT cell, or a tumor-infiltrating lymphocyte. In some aspects, the cell is a T cell. In some aspects, the cell is an alpha-beta T cell or a gamma-delta T cell. In some aspects, the cell is a gamma delta T cell comprises V delta 1. In some aspects, the cell is a gamma delta T cell comprises V delta 2.

[0165] In some aspects, the cell is a cell differentiated from an iPSC. In some aspects, the cell is an immune cell differentiated from an iPSC. In some aspects, the cell is a T cell differentiated from an iPSC. In some aspects, the cell is an NK cell differentiated from an iPSC. In some aspects, the cell is an NKT cell differentiated from an iPSC.

[0166] In some aspects, the cell comprises a CAR or an eTCR, wherein the CAR or the eTCR comprises an antigen-binding domain that specifically binds a tumor antigen. In some aspects, the tumor antigen comprises CD 19, CD20, ROR1, CD22, carcinoembryonic antigen, alphafetoprotein, CA-125, 5T4, MUC-1, epithelial tumor antigen, prostate-specific antigen, melanoma-associated antigen, mutated p53, mutated ras, HER2/Neu, folate binding protein, HIV-1 envelope glycoprotein gpl20, HIV-1 envelope glycoprotein gp41, GD2, CD123, CD33, CD138, CD23, CD30, CD56, c-Met, mesothelin, GD3, HERV-K, IL-llRalpha, kappa chain, lambda chain, CSPG4, ERBB2, EGFRvIII, VEGFR2, HER2-HER3 in combination, HER1-HER2 in combination, NY- ESO-1, synovial sarcoma X breakpoint 2 (SSX2), melanoma antigen (MAGE), melanoma antigen recognized by T cells 1 (MART-1), gplOO, prostate specific antigen (PSA), prostate specific membrane antigen (PSMA), prostate stem cell antigen (PSCA), GPC3, EpCAM, BCMA, GCC, ADGRE2, claudin (e.g., CLDN18.2) , B7H3 or any combination thereof.

[0167] In some aspects, the cell comprises a CAR, wherein the CAR comprises an antigen-binding domain that specifically binds mesothelin. In some aspects, the cell comprises an eTCR, wherein the eTCR comprises an antigen-binding domain that specifically binds mesothelin. In some aspects, the cell comprises an eTCR wherein the eTCR is a gamma-delta TCR. In some aspects, the cell comprises an eTCR wherein the eTCR is a g9d2TCR.

[0168] In some aspects, the cell comprises a CAR, wherein the CAR comprises an antigen-binding domain that specifically binds CD 19. In some aspects, the cell comprises an eTCR, wherein the eTCR comprises an antigen-binding domain that specifically binds CD 19.

[0169] In some aspects, the cell comprises a CAR, wherein the CAR comprises an antigen-binding domain that specifically binds BCMA. In some aspects, the cell comprises an eTCR, wherein the eTCR comprises an antigen-binding domain that specifically binds BCMA.

[0170] Some aspects of the present disclosure are directed to a population of cells comprising one or more modified cell disclosed herein. In some aspects, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% of the population of cells comprise the modifications or combination of modifications disclosed herein. In other words, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% of cells in a population of cells are modified cells described herein. In some aspects, at least about 25% of the population of cells comprises the modified cell disclosed herein. In some aspects, at least about 30% of the population of cells comprises the modified cell disclosed herein. In some aspects, at least about 35% of the population of cells comprises the modified cell disclosed herein. In some aspects, at least about 40% of the population of cells comprises the modified cell disclosed herein. In some aspects, at least about 45% of the population of cells comprises the modified cell disclosed herein. In some aspects, at least about 50% of the population of cells comprises the modified cell disclosed herein. In some aspects, at least about 55% of the population of cells comprises the modified cell disclosed herein. In some aspects, at least about 60% of the population of cells comprises the modified cell disclosed herein. In some aspects, at least about 65% of the population of cells comprises the modified cell disclosed herein. In some aspects, at least about 70% of the population of cells comprises the modified cell disclosed herein. In some aspects, at least about 75% of the population of cells comprises the modified cell disclosed herein.

II. B. Fas Constructs

[0171] Some aspects of the present disclosure are directed to modified and chimeric polypeptides comprising a Fas receptor extracellular domain. Further aspects are directed to a modified cell, e.g., immune cell, e.g., iCAR-T cell, comprising a modified or chimeric polypeptide comprising a Fas receptor extracellular domain (Fas ECD).

II.B.l. Fas-DN

[0172] In some aspects, the modified polypeptide is a dominant-negative Fas (Fas-DN). As such, some aspects of the present disclosure are directed to a modified cell, e.g., immune cell, e.g., iCAR-T cell, comprising a Fas-DN. In some aspects, the Fas-DN comprises an extracellular domain of a Fas receptor and a modification (e.g., a deletion or substitution) of one or more amino acids in the intracellular (cytoplasmic) domain of the Fas receptor. In some aspects, the Fas-DN comprises an extracellular domain of a Fas receptor and a deletion of one or more amino acids in the intracellular domain of a Fas receptor. In some aspects, the modification (e.g., deletion or substitution) is in one or more amino acids selected from residues 191-335 of the Fas sequence. In some aspects, the Fas-DN has decreased or no interaction with Fas-ligand. In some aspects, the Fas-DN is not capable of recruiting and/or activating caspase 8.

[0173] In some aspects, the Fas-DN comprises an extracellular domain of Fas receptor and a truncated intracellular domain of the Fas receptor. In some aspects, the truncated Fas receptor comprises a deletion of one or more amino acids selected from residues 230 to 312 of the canonical Fas receptor amino acid sequence (SEQ ID NO: 30). In some aspects, the truncated Fas receptor comprises a deletion of at least about 5, at least about 10, at least about 15, at least about 20, at least about 25, at least about 30, at least about

35, at least about 40, at least about 45, at least about 50, at least about 55, at least about

60, at least about 65, at least about 70, at least about 75, at least about 80, at least about

85, or at least about 90 amino acids from residues 230 to 312 of the canonical Fas receptor amino acid sequence (SEQ ID NO: 30). In some aspects, the truncated Fas receptor comprises a deletion of residues 230 to 305, residues 230 to 306, residues 230 to 307, residues 230 to 308, residues 230 to 309, residues 230 to 310, residues 230 to 311, residues 230 to 312, residues 231 to 312, residues 232 to 312, residues 233 to 312, residues 234 to 312, residues 235 to 312, residues 236 to 312, residues 237 to 312, residues 238 to 312, residues 239 to 312, or residues 240 to 312 of the canonical Fas receptor amino acid sequence (SEQ ID NO: 30). In some aspects, the truncated Fas receptor comprises a deletion residues 230 to 312 of the canonical Fas receptor amino acid sequence (SEQ ID NO: 30) and deletion of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten additional amino acids. In some aspects, the truncated Fas receptor comprises a deletion of residues 230 to 313, residues 230 to 314, residues 230 to 315, residues 230 to 316, residues 230 to 317, residues 229 to 312, residues 228 to 312, residues 227 to 312, residues 226 to 312, or residues 225 to 312 of the canonical Fas receptor amino acid sequence (SEQ ID NO: 30). In some aspects, the truncated Fas receptor comprises a deletion residues 230 to 312 of the canonical Fas receptor amino acid sequence (SEQ ID NO: 30).

[0174] In some aspects, the Fas-DN comprises an amino acid sequence having at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 22. In some aspects, the chimeric polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 27. In some aspects, the Fas-DN comprises the amino acid sequence set forth in SEQ ID NO: 27. In some aspects, the Fas-DN further comprises a signal peptide. In some aspects, the Fas-DN comprises a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 28. In some aspects, the Fas-DN comprises a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 29.

II.B.2. Fas-CD27

[0175] In some aspects, the chimeric polypeptide is a Fas-CD27 chimeric polypeptide (Fas-CD27). As such, some aspects of the present disclosure are directed to a modified cell, e.g, immune cell, e.g, iCAR-T cell, comprising a Fas-CD27. In some aspects, the chimeric polypeptide comprises a Fas extracellular domain linked to a CD27 intracellular domain or a portion thereof. In some aspects, the chimeric polypeptide comprises a Fas extracellular domain linked to a portion of a CD27 intracellular domain, wherein the portion of the CD27 intracellular domain retains one or more intracellular signaling function of the full-length CD27.

[0176] In some aspects, the Fas-CD27 polypeptide comprises an amino acid sequence having at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 22. In some aspects, the Fas-CD27 polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 22. In some aspects, the Fas-CD27 further comprises a signal peptide. In some aspects, the Fas-CD27 comprises a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 28. In some aspects, the Fas-CD27 comprises a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 29.

II.B.3. Fas-4-lBB

[0177] In some aspects, the chimeric polypeptide is a Fas-4-lBB chimeric polypeptide (Fas-4-lBB). As such, some aspects of the present disclosure are directed to a modified cell, e.g., immune cell, e.g., iCAR-T cell, comprising a Fas-4-lBB. In some aspects, the chimeric polypeptide comprises a Fas extracellular domain linked to a 4-1BB intracellular domain or a portion thereof. In some aspects, the chimeric polypeptide comprises a Fas extracellular domain linked to a portion of a 4- IBB intracellular domain, wherein the portion of the 4- IBB intracellular domain retains one or more intracellular signaling function of the full-length 4-1BB.

[0178] In some aspects, the Fas-4-lBB polypeptide comprises an amino acid sequence having at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 23. In some aspects, the Fas-4-lBB polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 23. In some aspects, the Fas-4-lBB further comprises a signal peptide. In some aspects, the Fas-4-lBB comprises a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 28. In some aspects, the Fas-4-lBB comprises a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 29.

II.B.4. Fas-OX40

[0179] In some aspects, the chimeric polypeptide is a Fas-OX40 chimeric polypeptide (Fas-OX40). As such, some aspects of the present disclosure are directed to a modified cell, e.g., immune cell, e.g., iCAR-T cell, comprising a Fas-OX40. In some aspects, the chimeric polypeptide comprises a Fas extracellular domain linked to an 0X40 intracellular domain or a portion thereof. In some aspects, the chimeric polypeptide comprises a Fas extracellular domain linked to a portion of an 0X40 intracellular domain, wherein the portion of the 0X40 intracellular domain retains one or more intracellular signaling function of the full-length 0X40.

[0180] In some aspects, the Fas-OX40 polypeptide comprises an amino acid sequence having at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 24. In some aspects, the chimeric polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 24.

[0181] In some aspects, the Fas-OX40 polypeptide comprises an amino acid sequence having at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 25. In some aspects, the chimeric polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 25. [0182] In some aspects, the Fas-OX40 polypeptide comprises an amino acid sequence having at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 26. In some aspects, the chimeric polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 26.

[0183] In some aspects, the Fas-OX40 further comprises a signal peptide. In some aspects, the Fas-OX40 comprises a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 28. In some aspects, the Fas-OX40 comprises a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 29.

II. C. CD58-Specific Guide RNAs

[0184] Some aspects of the present disclosure are directed to a guide RNA capable of hybridizing to a human CD58 gene. In some aspects, wherein the guide RNA hybridizes to a nucleotide sequence in Exon 3 of the endogenous CD58 gene. In some aspects, the guide RNA comprises a nucleotide sequence selected from SEQ ID NOs: 1-20 and 41.

[0185] In some aspects, the guide RNA comprises the nucleotide sequence set forth in SEQ ID NO: 1. In some aspects, the guide RNA consists of the nucleotide sequence set forth in SEQ ID NO: 1. In some aspects, the guide RNA comprises a nucleotide sequence that hybridizes to a nucleotide sequence that overlaps with a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 1.

[0186] In some aspects, the guide RNA comprises the nucleotide sequence set forth in SEQ ID NO: 2. In some aspects, the guide RNA consists of the nucleotide sequence set forth in SEQ ID NO: 2. In some aspects, the guide RNA comprises a nucleotide sequence that hybridizes to a nucleotide sequence that overlaps with a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 2.

[0187] In some aspects, the guide RNA comprises the nucleotide sequence set forth in SEQ ID NO: 3. In some aspects, the guide RNA consists of the nucleotide sequence set forth in SEQ ID NO: 3. In some aspects, the guide RNA comprises a nucleotide sequence that hybridizes to a nucleotide sequence that overlaps with a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 3.

[0188] In some aspects, the guide RNA comprises the nucleotide sequence set forth in SEQ ID NO: 4. In some aspects, the guide RNA consists of the nucleotide sequence set forth in SEQ ID NO: 4. In some aspects, the guide RNA comprises a nucleotide sequence that hybridizes to a nucleotide sequence that overlaps with a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 4.

[0189] In some aspects, the guide RNA comprises the nucleotide sequence set forth in SEQ ID NO: 5. In some aspects, the guide RNA consists of the nucleotide sequence set forth in SEQ ID NO: 5. In some aspects, the guide RNA comprises a nucleotide sequence that hybridizes to a nucleotide sequence that overlaps with a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 5.

[0190] In some aspects, the guide RNA comprises the nucleotide sequence set forth in SEQ ID NO: 6. In some aspects, the guide RNA consists of the nucleotide sequence set forth in SEQ ID NO: 6. In some aspects, the guide RNA comprises a nucleotide sequence that hybridizes to a nucleotide sequence that overlaps with a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 6.

[0191] In some aspects, the guide RNA comprises the nucleotide sequence set forth in SEQ ID NO: 7. In some aspects, the guide RNA consists of the nucleotide sequence set forth in SEQ ID NO: 7. In some aspects, the guide RNA comprises a nucleotide sequence that hybridizes to a nucleotide sequence that overlaps with a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 7.

[0192] In some aspects, the guide RNA comprises the nucleotide sequence set forth in SEQ ID NO: 8. In some aspects, the guide RNA consists of the nucleotide sequence set forth in SEQ ID NO: 8. In some aspects, the guide RNA comprises a nucleotide sequence that hybridizes to a nucleotide sequence that overlaps with a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 8.

[0193] In some aspects, the guide RNA comprises the nucleotide sequence set forth in SEQ ID NO: 9. In some aspects, the guide RNA consists of the nucleotide sequence set forth in SEQ ID NO: 9. In some aspects, the guide RNA comprises a nucleotide sequence that hybridizes to a nucleotide sequence that overlaps with a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 9.

[0194] In some aspects, the guide RNA comprises the nucleotide sequence set forth in SEQ ID NO: 10. In some aspects, the guide RNA consists of the nucleotide sequence set forth in SEQ ID NO: 10. In some aspects, the guide RNA comprises a nucleotide sequence that hybridizes to a nucleotide sequence that overlaps with a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 10. [0195] In some aspects, the guide RNA comprises the nucleotide sequence set forth in SEQ ID NO: 11. In some aspects, the guide RNA consists of the nucleotide sequence set forth in SEQ ID NO: 11. In some aspects, the guide RNA comprises a nucleotide sequence that hybridizes to a nucleotide sequence that overlaps with a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 11.

[0196] In some aspects, the guide RNA comprises the nucleotide sequence set forth in SEQ ID NO: 12. In some aspects, the guide RNA consists of the nucleotide sequence set forth in SEQ ID NO: 12. In some aspects, the guide RNA comprises a nucleotide sequence that hybridizes to a nucleotide sequence that overlaps with a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 12.

[0197] In some aspects, the guide RNA comprises the nucleotide sequence set forth in SEQ ID NO: 13. In some aspects, the guide RNA consists of the nucleotide sequence set forth in SEQ ID NO: 13. In some aspects, the guide RNA comprises a nucleotide sequence that hybridizes to a nucleotide sequence that overlaps with a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 13.

[0198] In some aspects, the guide RNA comprises the nucleotide sequence set forth in SEQ ID NO: 14. In some aspects, the guide RNA consists of the nucleotide sequence set forth in SEQ ID NO: 14. In some aspects, the guide RNA comprises a nucleotide sequence that hybridizes to a nucleotide sequence that overlaps with a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 14.

[0199] In some aspects, the guide RNA comprises the nucleotide sequence set forth in SEQ ID NO: 15. In some aspects, the guide RNA consists of the nucleotide sequence set forth in SEQ ID NO: 15. In some aspects, the guide RNA comprises a nucleotide sequence that hybridizes to a nucleotide sequence that overlaps with a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 15.

[0200] In some aspects, the guide RNA comprises the nucleotide sequence set forth in SEQ ID NO: 16. In some aspects, the guide RNA consists of the nucleotide sequence set forth in SEQ ID NO: 16. In some aspects, the guide RNA comprises a nucleotide sequence that hybridizes to a nucleotide sequence that overlaps with a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 16.

[0201] In some aspects, the guide RNA comprises the nucleotide sequence set forth in SEQ ID NO: 17. In some aspects, the guide RNA consists of the nucleotide sequence set forth in SEQ ID NO: 17. In some aspects, the guide RNA comprises a nucleotide sequence that hybridizes to a nucleotide sequence that overlaps with a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 17.

[0202] In some aspects, the guide RNA comprises the nucleotide sequence set forth in SEQ ID NO: 18. In some aspects, the guide RNA consists of the nucleotide sequence set forth in SEQ ID NO: 18. In some aspects, the guide RNA comprises a nucleotide sequence that hybridizes to a nucleotide sequence that overlaps with a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 18.

[0203] In some aspects, the guide RNA comprises the nucleotide sequence set forth in SEQ ID NO: 19. In some aspects, the guide RNA consists of the nucleotide sequence set forth in SEQ ID NO: 19. In some aspects, the guide RNA comprises a nucleotide sequence that hybridizes to a nucleotide sequence that overlaps with a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 19.

[0204] In some aspects, the guide RNA comprises the nucleotide sequence set forth in SEQ ID NO: 20. In some aspects, the guide RNA consists of the nucleotide sequence set forth in SEQ ID NO: 20. In some aspects, the guide RNA comprises a nucleotide sequence that hybridizes to a nucleotide sequence that overlaps with a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 20.

[0205] In some aspects, the guide RNA comprises the nucleotide sequence set forth in SEQ ID NO: 41. In some aspects, the guide RNA consists of the nucleotide sequence set forth in SEQ ID NO: 41. In some aspects, the guide RNA comprises a nucleotide sequence that hybridizes to a nucleotide sequence that overlaps with a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 41.

[0206] Some aspects of the present disclosure are directed to a kit comprises (i) a guide RNA capable of hybridizing to a human CD58 gene disclosed herein and (ii) a gene editing tool. In some aspects, the gene editing tool comprises a CRISPR/Cas9, CRISPR/Casl2, TALEN, a zinc-finger endonuclease, or any combination thereof. In some aspects, the kit comprises (i) a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 1, and (ii) a CRISPR/Cas9. In some aspects, the kit comprises (i) a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 2, and (ii) a CRISPR/Cas9. In some aspects, the kit comprises (i) a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 3, and (ii) a CRISPR/Cas9. In some aspects, the kit comprises (i) a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 4, and (ii) a CRISPR/Cas9. In some aspects, the kit comprises (i) a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 5, and (ii) a CRISPR/Cas9. In some aspects, the kit comprises (i) a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 6, and (ii) a CRISPR/Cas9. In some aspects, the kit comprises (i) a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 7, and (ii) a CRISPR/Cas9. In some aspects, the kit comprises (i) a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 8, and (ii) a CRISPR/Cas9. In some aspects, the kit comprises (i) a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 9, and (ii) a CRISPR/Cas9. In some aspects, the kit comprises (i) a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 10, and (ii) a CRISPR/Cas9. In some aspects, the kit comprises (i) a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 11, and (ii) a CRISPR/Cas9. In some aspects, the kit comprises (i) a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 12, and (ii) a CRISPR/Cas9. In some aspects, the kit comprises (i) a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 13, and (ii) a CRISPR/Cas9. In some aspects, the kit comprises (i) a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 14, and (ii) a CRISPR/Cas9. In some aspects, the kit comprises (i) a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 15, and (ii) a CRISPR/Cas9. In some aspects, the kit comprises (i) a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 19, and (ii) a CRISPR/Cas9. In some aspects, the kit comprises (i) a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 17, and (ii) a CRISPR/Cas9. In some aspects, the kit comprises (i) a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 18, and (ii) a CRISPR/Cas9. In some aspects, the kit comprises (i) a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 19, and (ii) a CRISPR/Cas9. In some aspects, the kit comprises (i) a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 20, and (ii) a CRISPR/Cas9. In some aspects, the kit comprises (i) a guide RNA comprising the nucleotide sequence set forth in SEQ ID NO: 41, and (ii) a CRISPR/Cas9.

III. Methods of the Disclosure

III. A. Methods of Engineering

[0207] Some aspects of the present disclosure are directed to methods of engineering a human cell, comprising: (i) transfecting the cell with a nucleic acid encoding a CAR or an eTCR; (ii) mutating or deleting one or more endogenous genes encoding MHC-I human leukocyte antigens, wherein the mutation or deletion results in a reduced expression of MHC-I human leukocyte antigens relative to a wild-type cell of the same cell type; (iii) mutating or deleting one or more endogenous genes encoding MHC-II human leukocyte antigens, wherein the mutation or deletion results in a reduced expression of MHC-II human leukocyte antigens relative to a wild-type cell of the same cell type; and (iv) transfecting the cell with a nucleic acid encoding a polypeptide comprising Fas-DN, a Fas-CD27 chimeric polypeptide, a Fas-4-lBB chimeric polypeptide, a Fas-OX40 chimeric polypeptide, or any combination thereof.

[0208] Other aspects of the present disclosure are directed to methods of engineering a human cell, comprising: (i) inactivating one or more endogenous genes encoding MHC-I human leukocyte antigens, wherein the inactivation results in a reduced expression of MHC-I human leukocyte antigens relative to a wild-type cell of the same cell type; (ii) inactivating one or more endogenous genes encoding MHC-II human leukocyte antigens, wherein the inactivation results in a reduced expression of MHC-II human leukocyte antigens relative to a wild-type cell of the same cell type; and (iii) transfecting the cell with a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof.

[0209] Other aspects of the present disclosure are directed to methods of engineering a human cell, comprising: (i) inactivating one or more endogenous genes encoding MHC-I human leukocyte antigens, wherein the inactivation results in a reduced expression of MHC-I human leukocyte antigens relative to a wild-type cell of the same cell type; (ii) inactivating one or more endogenous genes encoding MHC-II human leukocyte antigens, wherein the inactivation results in a reduced expression of MHC-II human leukocyte antigens relative to a wild-type cell of the same cell type; and (iii) inactivating one or more endogenous genes encoding CD58, wherein the inactivation results in a reduced expression of CD58 relative to a wild-type cell of the same cell type.

[0210] Other aspects of the present disclosure are directed to methods of engineering a human cell, comprising: (i) inactivating one or more endogenous genes encoding MHC-I human leukocyte antigens, wherein the inactivation results in a reduced expression of MHC-I human leukocyte antigens relative to a wild-type cell of the same cell type; (ii) inactivating one or more endogenous genes encoding MHC-II human leukocyte antigens, wherein the inactivation results in a reduced expression of MHC-II human leukocyte antigens relative to a wild-type cell of the same cell type; (iii) inactivating one or more endogenous genes encoding CD58, wherein the inactivation results in a reduced expression of CD58 relative to a wild-type cell of the same cell type; and (iv) transfecting the cell with a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof.

[0211] In some aspects, the CAR or the eTCR comprises an antigen-binding domain that specifically binds a tumor antigen. In some aspects, the tumor antigen comprises CD 19, CD20, ROR1, CD22, carcinoembryonic antigen, alphafetoprotein, CA-125, 5T4, MUC-1, epithelial tumor antigen, prostate-specific antigen, melanoma-associated antigen, mutated p53, mutated ras, HER2/Neu, folate binding protein, HIV-1 envelope glycoprotein gpl20, HIV-1 envelope glycoprotein gp41, GD2, CD123, CD33, CD138, CD23, CD30, CD56, c- Met, mesothelin, GD3, HERV-K, IL-llRalpha, kappa chain, lambda chain, CSPG4, ERBB2, EGFRvIII, VEGFR2, HER2-HER3 in combination, HER1-HER2 in combination, NY-ESO-1, synovial sarcoma X breakpoint 2 (SSX2), melanoma antigen (MAGE), melanoma antigen recognized by T cells 1 (MART-1), gplOO, prostate specific antigen (PSA), prostate specific membrane antigen (PSMA), prostate stem cell antigen (PSCA), GPC3, EpCAM, BCMA, GCC, ADGRE, claudin (e g., CLDN18.2), B7H3 or any combination thereof.

[0212] In some aspects, the method comprises (i) transfecting the cell with a nucleic acid encoding a CAR; (ii) mutating or deleting one or more endogenous genes encoding MHC-I human leukocyte antigens, wherein the mutation or deletion results in a reduced expression of MHC-I human leukocyte antigens relative to a wild-type cell of the same cell type; (iii) mutating or deleting one or more endogenous genes encoding MHC-II human leukocyte antigens, wherein the mutation or deletion results in a reduced expression of MHC-II human leukocyte antigens relative to a wild-type cell of the same cell type; and (iv) transfecting the cell with a nucleic acid encoding a polypeptide comprising Fas-DN, a Fas-CD27 chimeric polypeptide, a Fas-4-lBB chimeric polypeptide, a Fas-OX40 chimeric polypeptide, or any combination thereof.

[0213] In some aspects, the method comprises (i) transfecting the cell with a nucleic acid encoding an eTCR; (ii) mutating or deleting one or more endogenous genes encoding MHC-I human leukocyte antigens, wherein the mutation or deletion results in a reduced expression of MHC-I human leukocyte antigens relative to a wild-type cell of the same cell type; (iii) mutating or deleting one or more endogenous genes encoding MHC-II human leukocyte antigens, wherein the mutation or deletion results in a reduced expression of MHC-II human leukocyte antigens relative to a wild-type cell of the same cell type; and (iv) transfecting the cell with a nucleic acid encoding a polypeptide comprising Fas-DN, a Fas-CD27 chimeric polypeptide, a Fas-4-lBB chimeric polypeptide, a Fas-OX40 chimeric polypeptide, or any combination thereof.

[0214] In some aspects, the method comprises (i) transfecting the cell with a nucleic acid encoding a CAR; (ii) mutating or deleting one or more endogenous genes encoding MHC-I human leukocyte antigens, wherein the mutation or deletion results in a reduced expression of MHC-I human leukocyte antigens relative to a wild-type cell of the same cell type; (iii) mutating or deleting one or more endogenous genes encoding MHC-II human leukocyte antigens, wherein the mutation or deletion results in a reduced expression of MHC-II human leukocyte antigens relative to a wild-type cell of the same cell type; and (iv) transfecting the cell with a nucleic acid encoding a polypeptide comprising Fas-DN. In some aspects, the method comprises (i) transfecting the cell with a nucleic acid encoding a CAR; (ii) mutating or deleting one or more endogenous genes encoding MHC-I human leukocyte antigens, wherein the mutation or deletion results in a reduced expression of MHC-I human leukocyte antigens relative to a wild-type cell of the same cell type; (iii) mutating or deleting one or more endogenous genes encoding MHC- II human leukocyte antigens, wherein the mutation or deletion results in a reduced expression of MHC-II human leukocyte antigens relative to a wild-type cell of the same cell type; and (iv) transfecting the cell with a nucleic acid encoding a polypeptide comprising a Fas-CD27 chimeric polypeptide, e.g., disclosed herein. In some aspects, the method comprises (i) transfecting the cell with a nucleic acid encoding a CAR; (ii) mutating or deleting one or more endogenous genes encoding MHC-I human leukocyte antigens, wherein the mutation or deletion results in a reduced expression of MHC-I human leukocyte antigens relative to a wild-type cell of the same cell type; (iii) mutating or deleting one or more endogenous genes encoding MHC-II human leukocyte antigens, wherein the mutation or deletion results in a reduced expression of MHC-II human leukocyte antigens relative to a wild-type cell of the same cell type; and (iv) transfecting the cell with a nucleic acid encoding a polypeptide comprising a Fas-4-lBB chimeric polypeptide, e.g., disclosed herein. In some aspects, the method comprises (i) transfecting the cell with a nucleic acid encoding a CAR; (ii) mutating or deleting one or more endogenous genes encoding MHC-I human leukocyte antigens, wherein the mutation or deletion results in a reduced expression of MHC-I human leukocyte antigens relative to a wild-type cell of the same cell type; (iii) mutating or deleting one or more endogenous genes encoding MHC-II human leukocyte antigens, wherein the mutation or deletion results in a reduced expression of MHC-II human leukocyte antigens relative to a wildtype cell of the same cell type; and (iv) transfecting the cell with a nucleic acid encoding a polypeptide comprising a Fas-OX40 chimeric polypeptide, e.g., disclosed herein.

[0215] In some aspects, the method comprises (i) transfecting the cell with a nucleic acid encoding an eTCR; (ii) mutating or deleting one or more endogenous genes encoding MHC-I human leukocyte antigens, wherein the mutation or deletion results in a reduced expression of MHC-I human leukocyte antigens relative to a wild-type cell of the same cell type; (iii) mutating or deleting one or more endogenous genes encoding MHC-II human leukocyte antigens, wherein the mutation or deletion results in a reduced expression of MHC-II human leukocyte antigens relative to a wild-type cell of the same cell type; and (iv) transfecting the cell with a nucleic acid encoding a polypeptide comprising Fas-DN. In some aspects, the method comprises (i) transfecting the cell with a nucleic acid encoding an eTCR; (ii) mutating or deleting one or more endogenous genes encoding MHC-I human leukocyte antigens, wherein the mutation or deletion results in a reduced expression of MHC-I human leukocyte antigens relative to a wild-type cell of the same cell type; (iii) mutating or deleting one or more endogenous genes encoding MHC- II human leukocyte antigens, wherein the mutation or deletion results in a reduced expression of MHC-II human leukocyte antigens relative to a wild-type cell of the same cell type; and (iv) transfecting the cell with a nucleic acid encoding a polypeptide comprising a Fas-CD27 chimeric polypeptide, e.g., disclosed herein. In some aspects, the method comprises (i) transfecting the cell with a nucleic acid encoding an eTCR; (ii) mutating or deleting one or more endogenous genes encoding MHC-I human leukocyte antigens, wherein the mutation or deletion results in a reduced expression of MHC-I human leukocyte antigens relative to a wild-type cell of the same cell type; (iii) mutating or deleting one or more endogenous genes encoding MHC-II human leukocyte antigens, wherein the mutation or deletion results in a reduced expression of MHC-II human leukocyte antigens relative to a wild-type cell of the same cell type; and (iv) transfecting the cell with a nucleic acid encoding a polypeptide comprising a Fas-4-lBB chimeric polypeptide, e.g., disclosed herein. In some aspects, the method comprises (i) transfecting the cell with a nucleic acid encoding an eTCR; (ii) mutating or deleting one or more endogenous genes encoding MHC-I human leukocyte antigens, wherein the mutation or deletion results in a reduced expression of MHC-I human leukocyte antigens relative to a wild-type cell of the same cell type; (iii) mutating or deleting one or more endogenous genes encoding MHC-II human leukocyte antigens, wherein the mutation or deletion results in a reduced expression of MHC-II human leukocyte antigens relative to a wildtype cell of the same cell type; and (iv) transfecting the cell with a nucleic acid encoding a polypeptide comprising a Fas-OX40 chimeric polypeptide, e.g., disclosed herein.

[0216] In some aspects, the method comprises (i) transfecting the cell with a nucleic acid encoding an eTCR; (ii) mutating or deleting one or more endogenous genes encoding B2M, wherein the mutation or deletion results in a reduced expression of B2M relative to a wild-type cell of the same cell type; (iii) mutating or deleting CIITA, wherein the mutation or deletion results in a reduced expression of MHC-II human leukocyte antigens relative to a wild-type cell of the same cell type; and (iv) transfecting the cell with a nucleic acid encoding a polypeptide comprising Fas-DN.

[0217] In some aspects, the method comprises (i) transfecting the cell with a nucleic acid encoding an eTCR; (ii) mutating or deleting one or more endogenous genes encoding B2M, wherein the mutation or deletion results in a reduced expression of B2M relative to a wild-type cell of the same cell type; (iii) mutating or deleting CIITA, wherein the mutation or deletion results in a reduced expression of MHC-II human leukocyte antigens relative to a wild-type cell of the same cell type; and (iv) transfecting the cell with a nucleic acid encoding a polypeptide comprising a Fas-CD27 chimeric polypeptide, e.g., disclosed herein.

[0218] In some aspects, the method comprises (i) transfecting the cell with a nucleic acid encoding an eTCR; (ii) mutating or deleting one or more endogenous genes encoding B2M, wherein the mutation or deletion results in a reduced expression of B2M relative to a wild-type cell of the same cell type; (iii) mutating or deleting CIITA, wherein the mutation or deletion results in a reduced expression of MHC-II human leukocyte antigens relative to a wild-type cell of the same cell type; and (iv) transfecting the cell with a nucleic acid encoding a polypeptide comprising a Fas-4-lBB chimeric polypeptide, e.g., disclosed herein. [0219] In some aspects, the method comprises (i) transfecting the cell with a nucleic acid encoding an eTCR; (ii) mutating or deleting one or more endogenous genes encoding B2M, wherein the mutation or deletion results in a reduced expression of B2M relative to a wild-type cell of the same cell type; (iii) mutating or deleting CIITA, wherein the mutation or deletion results in a reduced expression of MHC-II human leukocyte antigens relative to a wild-type cell of the same cell type; and (iv) transfecting the cell with a nucleic acid encoding a polypeptide comprising a Fas-OX40 chimeric polypeptide, e.g., disclosed herein.

[0220] In some aspects, the method further comprises mutating or deleting one or more endogenous genes encoding CD58, wherein the mutation or deletion results in a reduced expression of CD58 relative to a wild-type cell of the same cell type. In some aspects, the mutation or deletion of the one or more endogenous genes encoding CD58 comprises modifying the CD58 gene using a gene editing tool. In some aspects, the gene editing tool comprises a CRISPR/Cas9, CRISPR/Casl2, TALEN, a zinc-finger endonuclease, or any combination thereof. In some aspects, the one or more guide sequences hybridize to a nucleotide sequence in Exon 3 of the endogenous CD58 gene. In some aspects, the one or more guide sequences comprise a nucleotide sequence selected from SEQ ID NOs: 1-20 and 41. In some aspects, the guide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 1. In some aspects, the guide sequence consists of the nucleotide sequence set forth in SEQ ID NO: 1. In some aspects, the guide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 2. In some aspects, the guide sequence consists of the nucleotide sequence set forth in SEQ ID NO: 2. In some aspects, the guide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 3. In some aspects, the guide sequence consists of the nucleotide sequence set forth in SEQ ID NO: 3. In some aspects, the guide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 4. In some aspects, the guide sequence consists of the nucleotide sequence set forth in SEQ ID NO: 4. In some aspects, the guide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 5. In some aspects, the guide sequence consists of the nucleotide sequence set forth in SEQ ID NO: 5. In some aspects, the guide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 6. In some aspects, the guide sequence consists of the nucleotide sequence set forth in SEQ ID NO: 6. In some aspects, the guide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 7. In some aspects, the guide sequence consists of the nucleotide sequence set forth in SEQ ID NO: 7. In some aspects, the guide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 8. In some aspects, the guide sequence consists of the nucleotide sequence set forth in SEQ ID NO: 8. In some aspects, the guide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 9. In some aspects, the guide sequence consists of the nucleotide sequence set forth in SEQ ID NO: 9. In some aspects, the guide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 10. In some aspects, the guide sequence consists of the nucleotide sequence set forth in SEQ ID NO: 10. In some aspects, the guide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 11. In some aspects, the guide sequence consists of the nucleotide sequence set forth in SEQ ID NO: 11. In some aspects, the guide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 12. In some aspects, the guide sequence consists of the nucleotide sequence set forth in SEQ ID NO: 12. In some aspects, the guide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 13. In some aspects, the guide sequence consists of the nucleotide sequence set forth in SEQ ID NO: 13. In some aspects, the guide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 14. In some aspects, the guide sequence consists of the nucleotide sequence set forth in SEQ ID NO: 14. In some aspects, the guide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 15. In some aspects, the guide sequence consists of the nucleotide sequence set forth in SEQ ID NO: 15. In some aspects, the guide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 16. In some aspects, the guide sequence consists of the nucleotide sequence set forth in SEQ ID NO: 16. In some aspects, the guide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 17. In some aspects, the guide sequence consists of the nucleotide sequence set forth in SEQ ID NO: 17. In some aspects, the guide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 18. In some aspects, the guide sequence consists of the nucleotide sequence set forth in SEQ ID NO: 18. In some aspects, the guide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 19. In some aspects, the guide sequence consists of the nucleotide sequence set forth in SEQ ID NO: 19. In some aspects, the guide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 20. In some aspects, the guide sequence consists of the nucleotide sequence set forth in SEQ ID NO: 20. In some aspects, the guide sequence consists of the nucleotide sequence set forth in SEQ ID NO: 41. [0221] In some aspects, the method further comprises mutating or deleting one or more endogenous genes encoding PVR, wherein the mutation or deletion results in a reduced expression of PVR relative to a wild-type cell of the same cell type. Mutating or deleting one or more endogenous genes encoding PVR can be achieved using any method, including, but not limited to, the use of a gene editing tool, e.g., a gene editing tool disclosed herein.

[0222] In some aspects, the method further comprises transfecting the cells with a nucleic acid encoding an HLA-E polypeptide, e.g., an HLA-E polypeptide disclosed herein. In some aspects, the method further comprises modifying an endogenous gene encoding an HLA-E polypeptide, wherein the modification increases the expression of the endogenous HLA-E polypeptide. In some aspects, the method further comprises (i) transfecting the cells with a nucleic acid encoding an HLA-E polypeptide; and (ii) modifying an endogenous gene encoding an HLA-E polypeptide, wherein the modification increases the expression of the endogenous HLA-E polypeptide. In some aspects, the HLA-E polypeptide is a chimeric polypeptide comprising an HLA-E polypeptide linked to a B2M polypeptide.

[0223] In some aspects, the method further comprises transfecting the cells with a nucleic acid encoding an IL15 polypeptide, e.g., an IL15 polypeptide disclosed herein. In some aspects, the IL15 polypeptide is a membrane-bound IL15 polypeptide. In some aspects, the IL15 polypeptide is a membrane bound IL15/IL15Ralpha fusion polypeptide (mIL15/Ra). In some aspects, the IL15 polypeptide comprises an IL15 sushi domain/IL15Ra fusion polypeptide (sushi 15). In some aspects, the IL 15 polypeptide comprises a membrane bound-IL15/IL15Ra-LSP fusion (mIL15/Ra-LSP). In some aspects, the IL15 polypeptide comprises a short IL15 polypeptide (sIL15). In some aspects, the IL 15 polypeptide comprises a soluble IL 15 polypeptide.

[0224] In some aspects, the method further comprises transfecting the cells with a nucleic acid encoding a CCL19 polypeptide.

[0225] In some aspects, the endogenous gene can be inactivated by targeting for insertion an exogenous gene to a site of the endogenous gene. In some aspects, the exogenous gene comprises a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene, a nucleic acid encoding a promotor, a nucleic acid encoding a tag protein or any combination thereof. [0226] In some aspects, the promotor is a CAG promotor, an EFl alpha promotor, a CMV promotor, an hPGK promotor, or any combination thereof. In some aspects, the promotor comprises the nucleotide sequence set forth in SEQ ID NO: 37.

[0227] In some aspects, the tag protein is LNGFR or truncated LNGFR (e.g., delta LNGFR). In some aspects, the tag protein is EGFR. In some aspects, the tag protein is truncated EGFR (tEGFR).

[0228] In some embodiments, the exogenous gene to be inserted to the site of the endogenouse gene comprises HLA-E and CAG promotor. In some embodiments, the exogenous gene to be inserted to the site of the endogenouse gene comprises FasDN and CAG promotor. In some embodiments, the exogenous gene to be inserted to the site of the endogenouse gene comprises HLA-E, HSV-TK, FasDN and CAG promotor. In some embodiments, the exogenous gene to be inserted to the site of the endogenouse gene comprises HLA-E, HSV-TK, FasDN, EFl alpha promotor and CAG promotor. In some embodiments, the exogenous gene to be inserted to the site of the endogenouse gene comprises HLA-E, HSV-TK, FasDN, LNGFR or CAG promotor.

[0229] In some embodiments, the exogenous gene is inserted to the site of B2M gene. In some embodiments, the exogenous gene is inserted to the site of CIITA gene. In some embodiments, the exogenous gene is inserted to the site of CD58 gene.

[0230] In some embodiments, the exogenous gene is a nuceic acid comprising a nucleic acid sequence selected from SEQ ID NO: 33-40.

[0231] In some aspects, the cell is an iPSC. In some aspects, the cell is a cell differentiated from an iPSC. In some aspects, the cell is an immune cell differentiated from an iPSC. In some aspects, the cell is a T cell differentiated from an iPSC. In some aspects, the cell is an NK cell differentiated from an iPSC. In some aspects, the cell is an NKT cell differentiated from an iPSC. When the cell is an iPSC, it can be used as a source not only for an immune cell but also for any other type of cells that can be differentiated from the iPSC. Such differentiated cells, which have reduced in vivo immunogenicity, are useful for any cell therapies that comprise administering or transplanting such cells to a patient.

III.B . Methods of Treatment

[0232] Some aspects of the present disclosure are directed to methods of treating a disease or condition in a subject in need thereof comprising administering to the subject a composition disclosed herein. In some aspects, the method comprises administering a modified or engineered cell disclosed herein. In some aspects, the method comprises administering a population of cells disclosed herein. In some aspects, the method comprises administering a composition comprising a Fas construct disclosed herein. In some aspects, the method comprises administering a composition comprising a CD58- specific guide RNA.

[0233] In some aspects, the disease or condition comprises a cancer, e.g., the subject is afflicted with a cancer. In some aspects, the cancer comprises bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, Hodgkin's Disease, non-Hodgkin's lymphoma, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, chronic or acute leukemia, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, solid tumors of childhood, lymphocytic lymphoma, cancer of the bladder, cancer of the kidney or ureter, carcinoma of the renal pelvis, neoplasm of the central nervous system (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid cancer, squamous cell cancer, T-cell lymphoma, environmentally induced cancers including those induced by asbestos, or any combination thereof. In some aspects, the cancer is locally advanced. In some aspects, the cancer is metastatic. In some aspects, the cancer is refractory. In some aspects, the cancer is relapsed. In some aspects, the cancer is refractory or relapsed following one or more prior anti-cancer therapy. In some aspects, the one or more prior anti-cancer therapy comprises a standard of care therapy.

[0234] In some aspects, the compositions disclosed herein are administered in combination with an additional anti-cancer therapy. In some aspects, the additional anticancer therapy comprises a chemotherapy, an immunotherapy, a radiotherapy, a surgery, or any combination thereof. In some aspects, the additional anti-cancer therapy comprises a chemotherapy. In some aspects, the additional anti-cancer therapy comprises an immune-checkpoint inhibitor. In some aspects, the additional anti-cancer therapy comprises a PD-1 antagonist, a PD-L1 antagonist, a CTLA-4 antagonist, a LAG-3 antagonist, a GITR antagonist, or any combination thereof. In some aspects, the anticancer therapy comprises an antibody or antigen-binding portion thereof the specifically binds and inhibits PD-1. In some aspects, the anti-cancer therapy comprises an antibody or antigen-binding portion thereof the specifically binds and inhibits PD-L1.

[0235] In some aspects, the method further comprises pretreating the subject prior to administering the population of immune cells. In some aspects, the subject is administered a chemotherapy prior to administering the population of immune cells. In some aspects, the subject is administered an immuno-depleting chemotherapy prior to administering the population of immune cells. In some aspects, the immuno-depleting chemotherapy comprises cyclophosphamide, fludarabine, or both.

[0236] In some aspects, the method comprises administering to the subject (i) the population of expanded cells and (ii) a cytokine. In some aspects, the cytokine comprises IL-2, an analog thereof, a variant thereof, or a fragment thereof.

[0237] In some aspects, the cells of the present disclosure are administered to a subject at a dose of at least about 1 x 10⁶ cells, at least about 2 x 10⁶ cells, at least about 3 x 10⁶ cells, at least about 4 x 10⁶ cells, at least about 5 x 10⁶ cells, 1 x 10⁷ cells, at least about 2 x 10⁷ cells, at least about 3 x 10⁷ cells, at least about 4 x 10⁷ cells, at least about 5 x 10⁷ cells, 1 x 10⁸ cells, at least about 2 x 10⁸ cells, at least about 3 x 10⁸ cells, at least about 4 x 10⁸ cells, at least about 5 x 10⁸ cells, 1 x 10⁹ cells, at least about 2 x 10⁹ cells, at least about 3 x 10⁹ cells, at least about 4 x 10⁹ cells, or at least about 5 x 10⁹ cells.

III.C. Cell Therapy Products

[0238] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells disclosed herein. Some aspects of the present disclosure are directed to a cell therapy comprising a population of modified cells disclosed herein. In some aspects, the population of cells is cryopreserved. Any method of cry opreservation of cells, e.g., immune cells, can be used in the methods and compositions disclosed herein. In some aspects, the cells are cryopreserved in the presence of DMSO. In some aspects, the cell therapy is cryopreserved to facilitate shipment of the cells.

[0239] The cell therapy and/or population of cells disclosed herein can be further formulated with one or more excipient. Any excipient that can preserve the cells can be used in the methods and compositions disclosed herein. In some aspects, the cell therapy and/or population of cells is formulated with one or more excipient that allows for cry opreservation of the cells, e.g., DMSO.

[0240] In some aspects, the population of cells comprises a plurality of modified cells disclosed herein and one or more additional cell. In some aspects, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% of the cells in the population of cells comprise a plurality of modified cells disclosed herein. In some aspects, at least about 10% of the cells in the population of cells comprise a plurality of modified cells disclosed herein. In some aspects, at least about 20% of the cells in the population of cells comprise a plurality of modified cells disclosed herein. In some aspects, at least about 25% of the cells in the population of cells comprise a plurality of modified cells disclosed herein. In some aspects, at least about 30% of the cells in the population of cells comprise a plurality of modified cells disclosed herein. In some aspects, at least about 40% of the cells in the population of cells comprise a plurality of modified cells disclosed herein. In some aspects, at least about 50% of the cells in the population of cells comprise a plurality of modified cells disclosed herein. In some aspects, at least about 60% of the cells in the population of cells comprise a plurality of modified cells disclosed herein. In some aspects, at least about 65% of the cells in the population of cells comprise a plurality of modified cells disclosed herein. In some aspects, at least about 70% of the cells in the population of cells comprise a plurality of modified cells disclosed herein. In some aspects, at least about 75% of the cells in the population of cells comprise a plurality of modified cells disclosed herein. In some aspects, at least about 80% of the cells in the population of cells comprise a plurality of modified cells disclosed herein. In some aspects, at least about 90% of the cells in the population of cells comprise a plurality of modified cells disclosed herein.

[0241] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD 19. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD 19. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD 19.

[0242] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD20. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD20. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD20.

[0243] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds ROR1. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds ROR1. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds R0R1.

[0244] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD22. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD22. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD22.

[0245] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds carcinoembryonic antigen. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigenbinding domain that specifically binds carcinoembryonic antigen. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA- E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds carcinoembryonic antigen.

[0246] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds alphafetoprotein. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds alphafetoprotein. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds alphafetoprotein.

[0247] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CA-125. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CA-125. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CA-125.

[0248] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds 5T4. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA- E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds 5T4. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds 5T4.

[0249] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds MUC-1. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds MUC-1. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds MUC-1.

[0250] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds epithelial tumor antigen. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen- binding domain that specifically binds epithelial tumor antigen. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA- E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds epithelial tumor antigen.

[0251] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds prostate-specific antigen. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigenbinding domain that specifically binds prostate-specific antigen. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA- E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds prostate-specific antigen.

[0252] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds melanoma-associated antigen. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigenbinding domain that specifically binds melanoma-associated antigen. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA- E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds melanoma-associated antigen.

[0253] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds mutated p53. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds mutated p53. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds mutated p53.

[0254] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds mutated ras. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds mutated ras. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds mutated ras.

[0255] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds HER2/Neu. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds HER2/Neu. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds HER2/Neu.

[0256] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds folate binding protein. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigenbinding domain that specifically binds folate binding protein. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds folate binding protein.

[0257] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds HIV-1 envelope glycoprotein gpl20. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds HIV-1 envelope glycoprotein gpl20. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds HIV-1 envelope glycoprotein gpl20.

[0258] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds HIV-1 envelope glycoprotein gp41. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds HIV-1 envelope glycoprotein gp41. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds HIV-1 envelope glycoprotein gp41.

[0259] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds GD2. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA- E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds GD2. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds GD2.

[0260] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD123. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD123. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD 123.

[0261] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD33. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD33. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD33.

[0262] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD138. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD138. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD 138.

[0263] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD23. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD23. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD23. [0264] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD30. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD30. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD30.

[0265] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD56. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD56. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CD56.

[0266] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds c-Met. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds c-Met. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds c-Met.

[0267] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds mesothelin. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds mesothelin. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds mesothelin.

[0268] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds GD3. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA- E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds GD3. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds GD3.

[0269] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds HERV-K. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds HERV-K. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds HERV-K.

[0270] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds IL-llRalpha. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds IL-llRalpha. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds IL- HRalpha.

[0271] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds IL- kappa chain. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds kappa chain. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds kappa chain.

[0272] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds lambda chain. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds lambda chain. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds lambda chain.

[0273] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CSPG4. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CSPG4. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds CSPG4.

[0274] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds ERBB2. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds ERBB2. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds ERBB2.

[0275] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds EGFRvIII. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds EGFRvIII. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds EGFRvIII.

[0276] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds VEGFR2. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds VEGFR2. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds VEGFR2.

[0277] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds HER2-HER3 in combination. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigenbinding domain that specifically binds HER2-HER3 in combination. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA- E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds HER2-HER3 in combination.

[0278] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds HER1-HER2. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds HER1-HER2. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds HER1- HER2.

[0279] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds NY-ESO-1. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds NY-ESO-1. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds NY-ESO- 1.

[0280] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds SSX2. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds SSX2. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds SSX2.

[0281] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds MAGE. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds MAGE. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds MAGE.

[0282] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds MART-1. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds MART-1. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds MART-1.

[0283] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds gplOO. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds gplOO. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds gplOO.

[0284] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds PSA. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA- E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds PSA. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds PSA.

[0285] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds PSMA. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds PSMA. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds PSMA.

[0286] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds PSCA. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds PSCA. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds PSCA. [0287] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds GPC3. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds GPC3. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds GPC3.

[0288] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds EpCAM. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds EpCAM. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds EpCAM.

[0289] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds BCMA. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds BCMA. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds BCMA.

[0290] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds GCC. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds GCC. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds GCC.

[0291] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds ADGRE2. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds ADGRE2. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds ADGRE2.

[0292] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds claudin. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds claudin. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominantnegative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds claudin.

[0293] Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type or relative to the cell prior to the modification; (iii) a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type or relative to the cell prior to the modification, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds B7H3. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (iv) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds B7H3. Some aspects of the present disclosure are directed to a population of cells comprising a plurality of modified cells comprising: (i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA); (ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); (iii) inactivated endogenous gene encoding CD58; (iv) a nucleic acid encoding a dominant- negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof; and (v) a chimeric antigen receptor comprising an antigen-binding domain that specifically binds B7H3.

[0294] The practice of the present disclosure will employ, unless otherwise indicated, conventional techniques of cell biology, cell culture, molecular biology, transgenic biology, microbiology, recombinant DNA, and immunology, which are within the skill of the art. Such techniques are explained fully in the literature. See, for example, Sambrook et al., ed. (1989) Molecular Cloning A Laboratory Manual (2nd ed.; Cold Spring Harbor Laboratory Press); Sambrook et al., ed. (1992) Molecular Cloning: A Laboratory Manual, (Cold Springs Harbor Laboratory, NY); D. N. Glover ed., (1985) DNA Cloning, Volumes I and II; Gait, ed. (1984) Oligonucleotide Synthesis; Mullis et al. U.S. Pat. No. 4,683,195; Hames and Higgins, eds. (1984) Nucleic Acid Hybridization; Hames and Higgins, eds. (1984) Transcription And Translation; Freshney (1987) Culture Of Animal Cells (Alan R. Liss, Inc.); Immobilized Cells And Enzymes (IRL Press) (1986); Perbal (1984) A Practical Guide To Molecular Cloning; the treatise, Methods In Enzymology (Academic Press, Inc., N.Y.); Miller and Calos eds. (1987) Gene Transfer Vectors For Mammalian Cells, (Cold Spring Harbor Laboratory); Wu et al., eds., Methods In Enzymology, Vols. 154 and 155; Mayer and Walker, eds. (1987) Immunochemical Methods In Cell And Molecular Biology (Academic Press, London); Weir and Blackwell, eds., (1986) Handbook Of Experimental Immunology, Volumes I-FV; Manipulating the Mouse Embryo, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., (1986));

Crooke, Antisense drug Technology: Principles, Strategies and Applications, 2^nd Ed. CRC Press (2007) and in Ausubel et al. (1989) Current Protocols in Molecular Biology (John Wiley and Sons, Baltimore, Md.).

[0295] All of the references cited above, as well as all references cited herein, are incorporated herein by reference in their entireties.

[0296] The following examples are offered by way of illustration and not by way of limitation. EXAMPLES

EXAMPLE 1

[0297] Isolation and Expansion of Peripheral Blood Mononuclear cell Derived Alloreactive T cells (allo-Tcells)

[0298] T cells were negatively selected via immunomagnetic bead separation from peripheral blood mononuclear cells (PBMCs) using the EasySep Human T Cell Isolation Kit (Stemcell Technologies). Anti-mesothelin (MESO) chimeric antigen receptor (iCAR) T cells, generated from modified T cells derived from induced pluripotent cells (iPCs), were irradiated at 25Gy for 4 cycles, meso iCAR-T cells were then labeled with carbonxyfluorescein diacetate succinimidyl ester (CFSE; Life Technologies) in the dark for 30 minutes at 37°C. The labeling reaction was quenched with 40 mL of complete media. Irradiated meso iCAR-T cells were then co-cultured with T cells in the presence of IL-2 at a concentration of 2 ng/mL for 10 days. Media was replaced on day 4 and day 7 of culture. Alloreactive-T (allo-T) cells were then banked and cryopreserved.

[0299] In vitro Alloreactive T cell (allo-Tcell) Activation and Cytotoxicity assay

[0300] The allo-Tcells and meso iCAR-T cells were co-cultured to assess the killing of meso iCAR-T cells by allo-T cells. Wild type (WT) meso iCAR-T cells and MHC-I and MHC-II double knock-out (dKO) meso iCAR-T cells were labeled with cell trace violet (CTV; Life Technologies) in the dark for 30 minutes at 37°C. Approximately IxlO⁴ CTV- stained meso iCAR-T cells were co-cultured with allo-T cells either HLA matched or mis-matched in a 96-well plate at 4: 1, 2: 1, 1 : 1, and 0: 1 effectortarget (E:T) ratio for 24 hours at 37°C in a humidified 5% CO2 incubator. Co-cultured cells were centrifuged at 350 g for 5 minutes and then washed with PBS. Cells were then incubated with fixable live dead dye for 20 minutes at room temperature. Following incubation, cells were then washed with flow cytometry (FACS) buffer.

[0301] Flow Cytometry

[0302] Co-cultured allo-T cells and WT or dKO meso iCAR-T cells were stained with anti-CTV and anti-CD25 and run on FACS Canto and analyzed using FlowJo software. The activation status of allo-T cells was determined by detecting the CD25 activation marker (FIG. 1 A). The allo-Tcells were activated, and showed CD25 expression in PBMCs exposed to wild-type HLA mismatched iCAR-T cells (FIG. 1 A) but not HLA matched iCAR-T cells (FIG. 1C). Conversely, PBMCs cultutered with MHC-I/MHC-II dKO meso iCAR-T cells did not lead to activation of allo-T cells(FIGs. IB and 1C). Quantification of these representative plots are shown in FIG. IE.

[0303] In vitro allo-Tcell Cytotoxicity

[0304] Expanded and primed allo-T cells and meso iCAR-T cells were co-cultured to assess the allo-T cell-mediated killing of meso iCAR-T cells. Four distinct meso iCAR-T cell lines were used in this experiment: WT, P2M/CTIIA double KO, HLA-ABC/CTIIA double KO, and HLA-ABC/CIITA double KO with over expression of HLA-G and HLA- E. The meso iCAR-T cells were labeled with cell trace violet (CTV; Life Technologies) in the dark for 30 minutes at 37°C. Approximately, IxlO⁴ CTV-stained meso iCAR-T cells were co-cultured with HLA matched or HLA mis-matched allo-T cells in a 96-well plate at 2: 1, 1 : 1 effector Target (E:T) ratio for 24 hours at 37°C in a humidified 5% CO2 incubator. Co-cultured cells were centrifuged at 350 x g for 5 minutes and then washed with PBS. Cells were then incubated with fixable live dead dye for 20 minutes at room temperature. Following incubation, cells were then washed with flow cytometry (FACS) buffer. Flow cytometry was then performed on the cells. Hypo-immune edited meso iCAR-T cells including p2M/CTIIA double KO, HLA-ABC/CTIIA double KO, or HLA- ABC/CIITA double KO with overexpression of HLA-G and HLA-E had diminished recognition and killing by mis-matched allo-T cells (FIG. 2).

[0305] In vitro NK cell cytotoxicity assay

[0306] Natural killer (NK) cells derived from four different donors and meso iCAR-T cells were co-cultured to assess the killing of meso iCAR-T cells by NK cells. Meso iCAR-T cells were then labeled with carbonxyfluorescein diacetate succinimidyl ester (CFSE; Life Technologies) in the dark for 30 minutes at 37°C. The labeling reaction was quenched with 40 mL of complete media. Approximately, IxlO⁴ CFSE-stained meso iCAR-T cells were co-cultured with NK cells in a 96-well plate at 5: 1, 2: 1, 1 : 1, and 0: 1 effectortarget (E:T) ratio for 24 hours at 37°C in a humidified 5% CO2 incubator. Co- cultured cells were centrifuged at 350 g for 5 minutes and then washed with PBS. Cells were then incubated with fixable live dead dye for 20 minutes at room temperature. Following incubation, cells were then washed with flow cytometry (FACS) buffer.

[0307] An NK cell cytotoxicity assay was performed at 5: 1, 2:1, and 0: 1 iCAR-T effectortarget (E:T) ratio on different groups of hypo-immune edited meso iCAR-T cells including B2M/CTIIA double KO (DKO) (FIGs. 3 A, 3E, 31, and 3M), DKO and PVR KO (FIGs. 3B, 3F, 3J, and 3N), DKO and CD58 KO (FIGs. 3C, 3G, 3K, and 30), and DKO, CD58 KO, and PVR KO (FIGs. 3D, 3H, 3L, and 3P), for four different donors. These cells were compared to WT, untransduced, HLA-E overexpressed (OE), and FasDN OE or HLA-E OE meso iCAR-T cells.

[0308] An NK cell cytotoxicity assay was performed at 4: 1, 2: 1, and 1 : 1 E:T ratio on the following hypo-immune edited meso iCAR-T cells: B2M/CTIIA double KO (DKO); DKO and CD58 KO (TKO); TKO with overexpression of HLA-E and FasDN; TKO with overexpression of HLA-E and HSVTK; TKO with overexpression of HLA-E, FasDN, and HSVTK; and TKO with overexpression of HLA-E, FasBB, and HSVTK (FIG. 4). An NK cell cytotoxicity assay was performed at 5: 1 and 1.5: 1 effector Target (E:T) ratio on different groups of hypo-immune edited meso iCAR-T, including WT-Thyl. l, WT- CD47, WT-FasDN, WT-CD47-FasDN, B2M-Thyl .l, B2M-CD47, B2M-FasDN, B2M- CD47-FasDN, derived from different donors (FIGs. 5A-5C).

[0309] In vitro Tumor cell cytotoxicity assay

[0310] Meso iCAR-T cells and human-derived gastric cancer (GSU) cells, which have high expression levels of mesothelin, were co-cultured to assess the killing of GSU cancel cells by meso iCAR-T cells. Meso iCAR-T cells were co-cultured with GSU cells in a 96- well plate at 10: 1, 3:1, 1 : 1, and 0.3: 1 effectortarget (E:T) ratio for 24 hours at 37°C in a humidified 5% CO2 incubator. Co-cultured cells were then assessed using the cell titergio luminescent viability assay to determine the number of viable cells based on quantitation of ATP. The co-culture was incubated with cell titer-glo reagent for 15 minutes, which resulted in cell lysis and generation of a luminescent signal proportional to the amount of the ATP present. The luminescent signal was then measured using the Pherastar plate reader. WT- and hypo-immune-edited cells showed no difference in the effector functions of meso iCAR-T cells against GSU cancer cells (FIG. 6).

EXAMPLE 2

[0311] Reporter Description

[0312] Induced pluripotent derived stem cells were transduced with CAR containing firefly luciferase (Luc2). This reporter reacts with the substrate d-luciferin and creates a luminescent signal which can be observed between 600-800 nm, the optimal wavelength for bioluminescent imaging.

[0313] VIVOGLO™ Luciferin Preparation [0314] VIVOGLO™ Luciferin (Promega, Cat # P1043) was reconstituted in sterile DPBS (Gibco) at a concentration of 15 mg/mL and filtered through 0.2 uM filter. The resulting sterile luciferin was maintained at -20 °C. On the day of the study, aliquots of luciferin were removed from -20 °C storage, covered in foil to protect from light driven degradation, and maintained at room temperature. This was then transferred to the vivarium for injection.

[0315] NK Killing and Allo-T cell Rejection Mouse Background

[0316] Female 5 to 8-week-old NSG-MHC I/II DKO mutant mice edited to exhibit combined features of the severe combined immune deficiency mutation (scid), IL-2 receptor gamma chain deficiency, MHC class I molecule deficiency (H2-K and D), MHC class II molecule deficiency (IA), and a resistance to graft versus host disease (GVHD). This mouse line provides a model useful to study in vivo mechanisms of xenogeneic GVHD and to rapidly assess therapeutic agents.

[0317] GSULuc Efficacy Assay Mouse Background

[0318] Five to 8-week-old NOD.Cg-Prkdcscid I12rgtmlWjl/SzJ (NSG) mice were used for this experiment. These mice carry two mutations on the NOD/ShiLtJ genetic background: severe combined immune deficiency (scid) and a complete null allele of the IL-2 receptor common gamma chain (IL- . The scid mutation is in the DNA repair complex protein Prkdc and renders the mice B and T cell deficient. The IL- 2r ^lul1 mutation prevents cytokine signaling through multiple receptors, leading to a deficiency in functional NK cells. The severe immunodeficiency allows the mice to be humanized by engraftment of human CD34+ hematopoietic stem cells (HSC), peripheral blood mononuclear cells (PBMC), patient derived xenografts (PDX), or adult stem cells and tissues. The immunodeficient NSG mice enable research in human immune function, infectious disease, diabetes, oncology, and stem cell biology.

[0319] In vivo NK Cell Cytotoxicity Assay

[0320] Five to 8-week-old female NSG-MHC I/II DKO mice were weighed on day 0. Each mouse was intraperitoneally administered 200 uL of freshly prepared NK cells resuspended in PBS. After 1 hour, 200 uL of meso iCAR-T cells were administered intraperitoneally. One hour after administration of iCAR-T cells, 200 uL of d-luciferin substrate was injected intraperitoneally. After 9 minutes, animals were anesthetized and their ventral aspect was imaged on the IVIS Spectrum for a bioluminescent signal. This signal was then quantified as total flux (photons/second) within the region of interest (ROI). This process was repeated on Day 1, Day 3, and Day 6. Line graphs are presented as fold change over time. Fold change is defined as change in bioluminescent signal of each individual animal from baseline and represented as +/- SEM.

[0321] On Day 0, 1 hour before administration of meso iCAR-T cells, NSG-MHC I/II DKO animals were dosed with 2.0xl0⁶NK cells resuspended in DPBS. Following administration of NK cells, iCAR-T cells were prepared and resuspended in DPBS. Approximately, 1 hour after administration of NK cells, WT (positive control) and hypo- immune meso iCAR-T cells B2M/CIITA double KO (untransduced or UTD) (negative control), B2M/CIITA double KO with overexpression of HLA-E and FasDN, B2M/CIITA/PVR triple KO with overexpression of HLA-E and FasDN, B2M/CIITA/CD58 triple KO with overexpression of HLA-E and FasDN, and B2M/CIITA/CD58/PVR quadruple KO with overexpression of HLA-E and FasDN were administered at a dose of IxlO⁶ cells per animal. WT is defined as an induced pluripotent stem cell transduced with the mesothelin CAR. Normalized time course (FIG. 7A) and time courses using cells from two different donors (FIGs. 7B-7C) included the addition of HLA-E and FasDN overexpression in both the B2M/CIITA/CD58 KO and B2M/CIITA/PVR/CD58 KO.

[0322] An NK cell cytotoxicity assay in vivo was performed with mesoiCAR-T cells containing overexpression of FasBB and the kill switch HSVTK to assess the effect on persistence. The assay was performed on the following hypo-immune edited meso iCAR- T cell: B2M/CTIIA double KO (DKO), DKO and CD58 KO (TKO), TKO with overexpression of HLA-E and FasDN, TKO with overexpression of HLA-E and HSVTK, TKO with overexpression of HLA-E, FasDN, and HSVTK, and TKO with overexpression of HLA-E, FasBB, and HSVTK (FIG. 8A).

[0323] Alloreactive T (Allo-T) cell Rejection Assay

[0324] Five to 8-week-old female NSG-MHC I/II DKO mice were weighed on day 0 and intraperitoneally administered 200 uL of freshly prepared alloreactive T cells resuspended in PBS. After 1 hr, 200 uL of meso iCAR-T cells were administered intraperitoneally for each of WT, DKO-UTD, DKO with overexpression of HLA-E and FASDN, DKO and PVR KO with overexpression of HLA-E and FASDN, DKO and CD58 KO with overexpression of HLA-E and FASDN; and DKO and CD58/PVR KO with overexpression of HLA-E and FasDN. Approximately, 1 hour after administration of iCAR T cells, 200 uL of d-luciferin substrate was injected intraperitoneally. After 9 minutes, animals were anesthetized, and their ventral aspect was imaged on the IVIS Spectrum for a bioluminescent signal. This signal was then quantified as total flux (photons/second) within the region of interest (ROI). This process was repeated on Day 1, Day 3, and Day 6 (FIG. 8B).

[0325] GSU Efficacy model

[0326] Five to 8-week-old female NSG mice were intraperitoneally administered 200 uL of IxlO⁶ freshly prepared GSULuc cells resuspended in DPBS:Matrigel (1 : 1). On day 0, 200 uL IxlO⁶ iCAR T cells WT, DKO, DKO with overexpression of HLA-E1 and FasDN, and DKO and CD58 KO with overexpression of HLA-E1 and FasDN, and a negative control (PBS) were administered intraperitoneally. At day 7, 14, 21, 28, 35 and 42 animals were imaged, as described above (FIGs. 8C-8D).

EXAMPLE 3

[0327] Experimental design for FAS switch receptors

[0328] Meso iCAR-T cells were plated in a 96-well plate at a density of 10,000 cells/well. Four cell lines were used for these experiments, which included Meso iCAR-T cells with mIL15/Ra (5BB), dominant-negative Fas (dnFAS), Fas-BB (FAS-4-1BB), and Fas-Ox40 (FAS-OX40) (FIGs. 9A-9B). Super Fas-Ligand was added at 0.1 ng/ml, 1 ng/ml, 10 ng/ml, 100 ng/ml, or a PBS (negative control) for 5 days. After 5 days, cells were labeled with cell tracer violet dye for live/dead staining and counted using flow cytometry. Fas-Ox40 showed an increase cell proliferation after the addition of Super Fas-Ligand with a higher fold expansion of cells after activation with Fas-Ox40 compared to all other lines (FIGs. 10A-10F).

[0329] Long-term in vitro co-culture work flow

[0330] GSU target cells were plated at 40,000 cells/well on the morning of stimulation. The following effector cells were used for this assay, untransduced cells prepared with boost and non-boost protocols (UTD, UTD-Boost), meso-iCART cells with 5BB prepared with boost and non-boost protocols (5BB-Boost, 5BB), meso-iCART cells with 5BB and FasDN, and meso-iCART cells with 5BB and Fas-Ox40. In the boost protocol, frozen cells were recovered in IMDM medium with 15% FBS and IL2, IL7 and IL15 for 3 days. Cells were then activated on CD3/Retronectin coated plates in medium containing IL-2, IL-7, IL-15, IL-18, IL-21, Z-VAD inhibitor and CD30 agonistic antibody. Cells were then transferered to GREX flasks on Day3 and were harvested and banked on day6. Effector cells were added at 100,000 cells/well at an E:T ratio of 2.5: 1 and then co-cultured for 48 hours. After 48 hours, effector cells were collected for effector cell proliferation by counting viable CD3+ cells using flow cytometry (FIG. 11 A), and percent cytolysis of target cells were analyzed by cell titer gio assay (FIG. 1 IB). Simultaneously, 200 uL of collected effector cells were added to a fresh target cell plate seeded at 40,000 cells/well morning of stimulation and then co-cultured for 48 hours. The process was repeated for 2 weeks. At the end of 48 hours, effector cells were analyzed using the methods described above.

[0331] In vivo GSU Efficacy Model

[0332] Eight to 10-week-old female NSG mice were intraperitoneally administered 100 uL of IxlO⁶ freshly prepared GSULuc cells resuspended in DPBS:Matrigel (1 : 1). Mice were randomized on day 3 and meso-iCAR-T cells were injected on day 4. Group 1 was administered PBS as a negative control. Treatment mice were administered untransduced with mIL15/Ra (IL- 15 or 5BB in FIG. 12A-B), with 5BB and LNGFR, with 5BB and FasDN, with 5BB and Fas-41BB, with 5BB and Fas-CD27, or with 5BB and Fas-Ox40 (FIG. 12A). The Fas-Ox40 group showed the highest efficacy, which is highlighted in FIGs. 12B and 12C.

EXAMPLE 4

[0333] Modification of iPS cells

[0334] iPS cells were modified by electroporation (EP). Ribonucleoprotein (RNP) complex was generated by mixing Cas9 and gRNA for the site of B2M exon 1, CIITA exon 3 and CD58 exon 3 for knock-out (KO) of the genes. After RNP formation, plasmids for 1) HLA-E, 2) FasDN, and 3) HSVTK-tEGFR each were constructed with homology arms corresponding to each KI site generated by the KO of the genes in the aforementioned procedure, and a CAG promotor for the expression of each protein, and the resulting solution was subsequently mixed with iPSCs resuspended in buffer for knock-in (KI) of the genes. The KO/ KI combinations of interest are as follows: 1) B2M KO/CAG-HLA-E KI and, 2) CIITA KO/CAG-FASDN KI, or 3) CIITA KO/CAG- HSVTK-tEGFR KI, or 4) B2M KO/CAG-HLA-E-HSVTK KI, and 5) CD58 KO. EP was performed and afterwards, cells were plated in 6 well plates. Two weeks post-EP, modified cells were enriched for the KI surface marker by magnetic bead sorting. One week post-sorting, modified cells were assessed for KI population percentage using flow cytometry.

[0335] Modification of iT or iCART cells

[0336] iT or iCART cells were modified by electroporation (EP). Ribonucleoprotein (RNP) complex was generated by mixing Cas9 and gRNA for the site of B2M, CIITA, CD58, PVR or HLA-ABC for knock-out (KO) of the genes sequentially or at the same time. After RNP formation, EP was performed, and cells were plated in 6 well plates for recovery culture. For KI, edited KO iT or iCART cells were transduced with gamma retro virus containing HLA-E, or HLA-E-FASDN, with and without HSVTK or HLA-G and HLA-E, or FASDN, with and without CD47. Modified cells were enriched for the KO/KI surface marker by magnetic bead sorting. One to two weeks post-sorting, modified cells were assessed for KO/KI population percentage using flow cytometry.

[0337] Flow Cytometry

[0338] iPSCs modified with 1) B2M KO/HLA-E KI or 2) CIITA KO/FasDN KI or 3) CIITA KO/CAG-HSVTK-tEGFR KI were assessed by flow cytometry to determine the population expressing cell-surface KI protein. Cells were stained with an anti-HLA-E, anti-Fas (CD95), or anti-tEGFR antibody and Zombie Aqua live/dead stain, and assessed via flow cytometry using an Attune NxT cytometer and flowjo analysis software. The efficacy of the KO/KI protocol was determined independently for each KO/KI pair by examining the percentage of the live cell population positive for HLA-E in gene edited cells (FIG. 13 A) compared to wild type (WT) cells (FIG. 13B), and the live cell population positive for FasDN in gene edited cells (FIG. 13C) compared to WT cells (FIG. 13D), and the live cell population positive for tEGFR in gene edited cells (FIG. 13E) compared to WT cells (FIG 13F). Gene edited cells for HLA-E (FIG. 13A), FasDN (FIG. 13C), and HSVTK-tEGFR (FIG. 13E) showed successful KI of the genes of interest and subsequent expression. EXAMPLE 5

[0339] CD58 KO

[0340] The KO efficiencies of twenty CD58 gRNAs were evaluated in iCAR-T cells. A range of 10 pmol to 100 pmol of Ribonucleoprotein (RNP) complex was generated by mixing the appropriate amount of Cas9 and CD58 gRNAs. The resulting solution was subsequently mixed with iCAR-T cells resuspended in buffer for knock-out (KO) of the genes. Electroporation was performed, and modified iCAR-T cells were plated in 24-well plates after electroporation. CD58 knock-out percentages were assessed using flow cytometry 7 to 10 days post electroporation. The KO efficiencies are shown in Table 4 as A: > 75%, B: 65-75%, C: 50-65%, D: 30-50%, and E: < 30%.

Table 4: CD58 KO Efficiency

KO Efficiency is shown as A: > 75%, B: 65-75%, C: 50-65%, D: 30-50%, and E: < 30%. [0341] It is to be appreciated that the Detailed Description section, and not the Summary and Abstract sections, is intended to be used to interpret the claims. The Summary and Abstract sections may set forth one or more but not all exemplary embodiments of the present disclosure as contemplated by the inventor(s), and thus, are not intended to limit the present disclosure and the appended claims in any way.

[0342] The foregoing description of the specific embodiments will so fully reveal the general nature of the disclosure that others can, by applying knowledge within the skill of the art, readily modify and/or adapt for various applications such specific embodiments, without undue experimentation, without departing from the general concept of the present disclosure. Therefore, such adaptations and modifications are intended to be within the meaning and range of equivalents of the disclosed embodiments, based on the teaching and guidance presented herein. It is to be understood that the phraseology or terminology herein is for the purpose of description and not of limitation, such that the terminology or phraseology of the present specification is to be interpreted by the skilled artisan in light of the teachings and guidance.

[0343] The breadth and scope of the present disclosure should not be limited by any of the above-described exemplary embodiments, but should be defined only in accordance with the following claims and their equivalents.

[0344] The contents of all cited references (including literature references, U.S. or foreign patents or patent applications, and websites) that are cited throughout this application are hereby expressly incorporated by reference as if written herein in their entireties for any purpose, as are the references cited therein. Where any inconsistencies arise, material literally disclosed herein controls.

[0345] While various specific aspects have been illustrated and described, the above specification is not restrictive. It will be appreciated that various changes can be made without departing from the spirit and scope of the invention(s). Many variations will become apparent to those skilled in the art upon review of this specification.

Claims

WHAT IS CLAIMED: A modified cell, comprising:

(i) a chimeric antigen receptor (CAR) and/or an exogenous T cell receptor (eTCR);

(ii) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type;

(iii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; and

(iv) a polypeptide comprising a dominant-negative Fas (Fas-DN), a Fas-CD27 chimeric polypeptide (Fas-CD27), a Fas-4-lBB chimeric polypeptide (Fas-BB), a Fas- 0X40 chimeric polypeptide (Fas-OX40), or any combination thereof. The cell of claim 1, wherein the MHC-I human leukocyte antigens are HLA- A, HLA-B and HLA-C. The cell of claim 1, wherein the MHC-II human leukocyte antigens are HLA-DP, HLA- DQ and HLA-DR. The cell of claim 1, wherein the reduced expression of the MHC-I human leukocyte antigens results from a mutation in, or a deletion of, one or more endogenous genes encoding beta-2-microglobulin (B2M). The cell of claim 1, wherein the reduced expression of the endogenous MHC-II human leukocyte antigens results from a mutation in, or a deletion of, the one or more endogenous genes encoding class II major histocompatibility complex transactivator (CIITA). The cell of claim 1, further comprising a reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type. The cell of claim 6, wherein the reduced expression of the CD58 results from a mutation in, or a deletion of, one or more endogenous genes encoding CD58. The cell of any one of claims 1 to 7, comprising a nucleic acid encoding Fas-DN. The cell of any one of claims 1 to 7, comprising a nucleic acid encoding Fas-CD27. The cell of any one of claims 1 to 7, comprising a nucleic acid encoding Fas-BB. The cell of any one of claims 1 to 7, comprising a nucleic acid encoding Fas-OX40. A modified cell comprising:

(i) a CAR or an eTCR;

(ii) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type;

(iii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; and

(iv) a Fas-DN. A cell comprising:

(i) a CAR or an eTCR;

(ii) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type;

(iii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; and

(iv) a Fas-CD27. A cell comprising:

(i) a CAR or an eTCR;

(ii) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; (iii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; and

(iv) a Fas-BB. A cell comprising:

(i) a CAR or an eTCR;

(ii) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type;

(iii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; and

(iv) a Fas-OX40. The cell of any one of claims 12 to 15, further comprising a reduced expression of endogenous CD58 relative to a wild-type cell of the same cell type. The cell of claim 16, wherein the reduced expression of the CD58 results from, a mutation in, or a deletion of, one or more endogenous genes encoding CD58. The cell of any one of claims 1 to 17, further comprising a reduced expression of endogenous poliovirus receptor (PVR) relative to a wild-type cell of the same cell type. The cell of claim 18, wherein the reduced expression of the PVR results from a mutation in, or a deletion of, one or more endogenous genes encoding PVR. The cell of any one of claims 1 to 19, further comprising an increased expression of HLA-E relative to a wild-type cell of the same cell type. The cell of claim 20, wherein the increased expression of the HLA-E results from (i) a transfection of a nucleic acid encoding an HLA-E polypeptide; (ii) a modification to an endogenous gene encoding HLA-E polypeptide, wherein the modification increases the expression of the endogenous HLA-E polypeptide; or; (iii) both (i) and (ii). The cell of claim 21, wherein the HLA-E polypeptide is a chimeric polypeptide comprising an HLA-E polypeptide linked to a human B2M polypeptide. The cell of any one of claims 1 to 22, further comprising a human interleukin 15 (IL 15) polypeptide. The cell of claim 23, wherein the human IL15 polypeptide is a membrane bound IL15/IL15Ralpha fusion polypeptide (mIL15/Ra). The cell of any one of claims 1 to 24, further comprising a human chemokine (C-C motif) ligand 19 (CCL19) polypeptide. The cell of any one of claims 1 to 25, wherein the cell is an immune cell, an induced pluripotent stem cell (iPSC) or a cell differentiated from iPSC. The cell of any one of claims 1 to 26, wherein the cell (i) is an immune cell differentiated from an iPSC or (ii) a hematopoietic stem cell differentiated from an iPSC. The cell of any one of claims 1 to 27, wherein the cell comprises a T cell, an NK cell, an NKT cell, or a tumor-infiltrating lymphocyte. The cell of any one of claims 1 to 28, wherein the CAR or the eTCR comprises an antigen-binding domain that specifically binds a tumor antigen. The cell of claim 29, wherein the tumor antigen comprises CD19, CD20, R0R1, CD22, carcinoembryonic antigen, alphafetoprotein, CA-125, 5T4, MUC-1, epithelial tumor antigen, prostate-specific antigen, melanoma-associated antigen, mutated p53, mutated ras, HER2/Neu, folate binding protein, HIV-1 envelope glycoprotein gpl20, HIV-1 envelope glycoprotein gp41, GD2, CD123, CD33, CD138, CD23, CD30, CD56, c-Met, mesothelin, GD3, HERV-K, IL-llRalpha, kappa chain, lambda chain, CSPG4, ERBB2, EGFRvIII, VEGFR2, HER2-HER3 in combination, HER1-HER2 in combination, NY- ESO-1, synovial sarcoma X breakpoint 2 (SSX2), melanoma antigen (MAGE), melanoma antigen recognized by T cells 1 (MART-1), gplOO, prostate specific antigen (PSA), prostate specific membrane antigen (PSMA), prostate stem cell antigen (PSCA), GPC3, EpCAM, BCMA, GCC, ADGRE, claudin, B7H3 or any combination thereof. The cell of any one of claims 1 to 30, comprising a CAR, wherein the CAR comprises an antigen-binding domain that specifically binds mesothelin. A method of engineering a human cell, comprising:

(i) introducing into the cell, a nucleic acid encoding a chimeric antigen receptor (CAR) or an engineered T cell receptor (eTCR);

(ii) inactivating one or more endogenous genes encoding MHC-I human leukocyte antigens (HLA), wherein the inactivation results in a reduced expression of MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type;

(iii) inactivating one or more endogenous genes encoding MHC-II human leukocyte antigens (HLA), wherein the inactivation results in a reduced expression of MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; and

(iv) introducing into the cell, a nucleic acid encoding a polypeptide comprising a dominant-negative Fas (Fas-DN), a Fas-CD27 chimeric polypeptide (Fas-CD27), a Fas- 4-1BB chimeric polypeptide (Fas-BB), a Fas-OX40 chimeric polypeptide (Fas-OX40), or any combination thereof. The method of claim 32, further comprising:

(v) inactivating one or more endogenous genes encoding CD58, wherein the inactivation results in a reduced expression of CD58 relative to a wild-type cell of the same cell type. The method of claim 33, wherein the inactivation of the one or more endogenous genes encoding CD58 comprises modifying the CD58 gene using a gene editing tool. The method of claim 33 or 34, wherein the gene editing tool comprises a CRISPR/Cas9, CRISPR/Casl2, TALEN, a zinc-finger endonuclease, or any combination thereof. The method of any one of claims 33 to 35, wherein the inactivation of the one or more endogenous genes encoding CD58 comprises introducing into the cell CRISPR/Cas9 and a polynucleotide sequence encoding one or more guide sequences that hybridize to one or more target sequences within the CD58 gene. The method of claim 36, wherein the one or more guide sequences hybridize to a nucleotide sequence in Exon 3 of the endogenous CD58 gene. The method of claim 36 or 37, wherein the one or more guide sequences comprise a nucleotide sequence selected from SEQ ID NOs: 1-20 and 41. The method of any one of claims 32 to 38, further comprising inactivating one or more endogenous genes encoding poliovirus receptor (PVR), wherein the inactivation results in a reduced expression of PVR relative to a wild-type cell of the same cell type. The method of any one of claims 32 to 39, further comprising (i) introducing into the cells, a nucleic acid encoding an HLA-E polypeptide; (ii) modifying an endogenous gene encoding an HLA-E polypeptide, wherein the modification increases the expression of the endogenous HLA-E polypeptide; or (iii) both (i) and (ii). The method of claim 40, wherein the HLA-E polypeptide is a chimeric polypeptide comprising an HLA-E polypeptide linked to a B2M polypeptide. The method of any one of claims 32 to 41, further comprising transfecting the cell with a nucleic acid encoding an interleukin 15 (IL15) polypeptide. The method of claim 42, wherein the IL15 polypeptide is a membrane bound IL15/IL15Ralpha fusion polypeptide (mIL15/Ra). The method of any one of claims 32 to 43, further comprising introducing into the cell a heterologous nucleic acid encoding a human chemokine (C-C motif) ligand 19 (CCL19) polypeptide. The method of any one of claims 32 to 44, wherein the cell is an immune cell, an induced pluripotent stem cell (iPSC) or a cell differentiated from iPSC. The method of any one of claims 32 to 45, wherein the cell is an immune cell or a hematopoietic stem cell differentiated from an iPSC. The method of any one of claims 32 to 46, wherein the cell comprises a T cell, an NK cell, an NKT cell, or a tumor-infiltrating lymphocyte. The method of any one of claims 32 to 47, wherein the CAR or the eTCR comprises an antigen-binding domain that specifically binds a tumor antigen. The method of claim 48, wherein the tumor antigen comprises CD 19, CD20, R0R1, CD22, carcinoembryonic antigen, alphafetoprotein, CA-125, 5T4, MUC-1, epithelial tumor antigen, prostate-specific antigen, melanoma-associated antigen, mutated p53, mutated ras, HER2/Neu, folate binding protein, HIV-1 envelope glycoprotein gpl20, HIV-1 envelope glycoprotein gp41, GD2, CD123, CD33, CD138, CD23, CD30, CD56, c- Met, mesothelin, GD3, HERV-K, IL-llRalpha, kappa chain, lambda chain, CSPG4, ERBB2, EGFRvIII, VEGFR2, HER2-HER3 in combination, HER1-HER2 in combination, NY-ESO-1, synovial sarcoma X breakpoint 2 (SSX2), melanoma antigen (MAGE), melanoma antigen recognized by T cells 1 (MART-1), gplOO, prostate specific antigen (PSA), prostate specific membrane antigen (PSMA), prostate stem cell antigen (PSCA), GPC3, BCMA, GCC, ADGRE, claudin or any combination thereof. A cell prepared according to the method of any one of claims 32 to 49. The cell of any one of claims 1 to 31 and 50, which has increased persistence in vivo relative to a wild-type cell of the same cell type. A population of cells comprising the cell of any one of claims 1 to 31, 50, and 51. A cell population, wherein at least 50% of the cells in the population comprises a cell of any one of claims 1 to 31, 50, and 51. A method of treating a subject in need thereof, comprising administering to the subject the cell of any one of claims 1 to 31, 50, and 51 or the population of cells of claim 52 or 53. The method of claim 54, wherein the subject is afflicted with a cancer. The method of claim 55, wherein the cancer comprises bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, Hodgkin's Disease, non-Hodgkin's lymphoma, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, chronic or acute leukemia, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, solid tumors of childhood, lymphocytic lymphoma, cancer of the bladder, cancer of the kidney or ureter, carcinoma of the renal pelvis, neoplasm of the central nervous system (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid cancer, squamous cell cancer, T-cell lymphoma, environmentally induced cancers including those induced by asbestos, or any combination thereof. A guide RNA capable of hybridizing to a human CD58 gene, comprising a nucleic acid sequence selected from SEQ ID NOs: 1-20 and 41. The guide RNA of claim 57, comprising the nucleic acid sequence set forth in SEQ ID NO: 1. The guide RNA of claim 57, consisting of the nucleic acid sequence set forth in SEQ ID NO: 1. The guide RNA of claim 57, comprising the nucleic acid sequence set forth in SEQ ID NO: 2. The guide RNA of claim 57, consisting of the nucleic acid sequence set forth in SEQ ID NO: 2. The guide RNA of claim 57, comprising the nucleic acid sequence set forth in SEQ ID NO: 3. The guide RNA of claim 57, consisting of the nucleic acid sequence set forth in SEQ ID NO: 3. The guide RNA of claim 57, comprising the nucleic acid sequence set forth in SEQ ID NO: 4. The guide RNA of claim 57, consisting of the nucleic acid sequence set forth in SEQ ID NO: 4. A method of inactivating a human CD 58 gene in a cell, comprising contacting the cell with the guide RNA of any one of claims 57 to 65 or a nucleic acid encoding the guide RNA and a DNA endonuclease or a nucleic acid encoding the DNA endonuclease. The method of claim 59, wherein the DNA endonuclease comprises CRISPR/Cas9. A dominant-negative Fas (Fas-DN), comprising an amino acid sequence set forth in SEQ ID NO: 27. A Fas-CD27 chimeric polypeptide (Fas-CD27), comprising an amino acid sequence set forth in SEQ ID NO: 22. A Fas-4-lBB chimeric polypeptide (Fas-4-lBB), comprising an amino acid sequence set forth in SEQ ID NO: 23. A Fas-OX40 chimeric polypeptide (Fas-OX40), comprising an amino acid sequence set forth in SEQ ID NO: 24, SEQ ID NO: 25, or SEQ ID NO: 26. The cell of claim 1, wherein the eTCR is a gamma-delta TCR. The cell of claim 72, wherein the gamma-delta TCR is V gamma 9-V delta 2 TCR (g9d2TCR). A modified cell comprising:

(i) reduced expression of endogenous MHC-I human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type;

(ii) reduced expression of endogenous MHC-II human leukocyte antigens (HLA) relative to a wild-type cell of the same cell type; and

(iii) expression of a Fas (Fas-DN), increased expression of HLA-E relative to a wild-type cell of the same cell type, a nucleic acid encoding a suicide gene or any combination thereof. A modified cell comprising:

(i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA);

(ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA); and (iii) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof. A modified cell comprising:

(i) inactivated endogenous gene encoding MHC-I human leukocyte antigens (HLA);

(ii) inactivated endogenous gene encoding MHC-II human leukocyte antigens (HLA);

(iii) inactivated endogenous gene encoding CD58; and

(iv) a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof. A method of engineering a human cell, comprising:

(i) inactivating one or more endogenous genes encoding MHC-I human leukocyte antigens, wherein the inactivation results in a reduced expression of MHC-I human leukocyte antigens relative to a wild-type cell of the same cell type;

(ii) inactivating one or more endogenous genes encoding MHC-II human leukocyte antigens, wherein the inactivation results in a reduced expression of MHC-II human leukocyte antigens relative to a wild-type cell of the same cell type; and

(iii) transfecting the cell with a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof. A method of engineering a human cell, comprising:

(i) inactivating one or more endogenous genes encoding MHC-I human leukocyte antigens, wherein the inactivation results in a reduced expression of MHC-I human leukocyte antigens relative to a wild-type cell of the same cell type;

(ii) inactivating one or more endogenous genes encoding MHC-II human leukocyte antigens, wherein the inactivation results in a reduced expression of MHC-II human leukocyte antigens relative to a wild-type cell of the same cell type; (iii) inactivating one or more endogenous genes encoding CD58, wherein the inactivation results in a reduced expression of CD58 relative to a wild-type cell of the same cell type; and

(iv) transfecting the cell with a nucleic acid encoding a dominant-negative Fas (Fas-DN), a nucleic acid encoding an exogenous HLA-E polypeptide, a nucleic acid encoding a suicide gene or any combination thereof.