Antigen-specific t cell compositions for treating cancer and methods of making the same

Abstract

Provided are T cell compositions and endothelin receptor antagonists for treating cancer and methods of making the same. In some embodiments, the present technology comprises methods of preventing or reducing a cancer in a subject in need thereof relative to a control, comprising administering to the subject (a) a T cell composition specific to at least a portion of one or more cancer antigens and (b) an endothelin receptor antagonist. In some embodiments, the methods further comprise administering (c) an immune checkpoint inhibitor to the subject.

Description

ANTIGEN-SPECIFIC T CELL COMPOSITIONS FOR TREATING CANCER AND METHODS OF MAKING THE SAMECROSS-REFERENCE TO RELATED APPLICATION(S)

[0001] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 713,535, filed October 29, 2024, the contents of which are incorporated herein by reference in their entirety.BACKGROUND

[0002] Immunotherapy aims to leverage the immune system to alter the course of diseases such as cancer. Cancer immunotherapy has evolved in various directions, including vaccines that boost immune responses against tumor antigens, ex vivo expansion of autologous cytotoxic T-cells with or without engineered receptors for tumor recognition, and monoclonal antibodies (mAb) to release inhibitory receptor-ligand interactions on immune cells, known as immune checkpoint blockade immunotherapy. In vitro preparation of T cells for adoptive cell transfer (ACT) of cancer allows for the selection of cells with high- avidity recognition of the tumor, expansion of tumor-specific T cells away from the suppressive properties of the tumor microenvironment, and manipulation of the T-cells to enhance their anti-tumor activity, such as the introduction of endogenous TCR or chimeric antigen receptors (CARs). ACT also provides the ability to manipulate the host microenvironment to increase favorable conditions for recruiting transferred T-cells.

[0003] Although ACT with autologous tumor-infiltrating lymphocytes (TILs) has been successful in metastatic melanoma, the overall success rate remains low. Despite the advancements, there remains a significant gap in the field, where some ACT therapies for solid tumors may fail to discriminate healthy cells from malignant cells. As such, immunotherapies having enhanced efficacy and cancer antigen recognition are needed.SUMMARY

[0004] The present technology comprises a method of preventing or reducing a cancer in a subject in need thereof relative to a control, comprising administering to the subject (a)a T cell composition specific to at least a portion of one or more cancer antigens; and (b) an endothelin receptor antagonist.

[0005] The present technology comprises a method of reducing a number of cancer cells in a subject in need thereof relative to a control, comprising administering to the subject (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; and (b) an endothelin receptor antagonist.

[0006] The present technology comprises a method of preventing or reducing a tumor volume in a subject in need thereof relative to a control, comprising administering to the subject (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; and (b) an endothelin receptor antagonist.

[0007] The present technology comprises a method of preventing or reducing cancer metastasis in a subject in need thereof relative to a control, comprising administering to the subject (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; and (b) an endothelin receptor antagonist.

[0008] The present technology comprises a method of preventing or reducing a cancer in a subject in need thereof relative to a control, comprising administering to the subject (a) a T cell composition specific to one or more cancer antigens; (b) an endothelin receptor antagonist; and (c) one or more immune checkpoint inhibitors.

[0009] The present technology comprises a method of preventing or reducing a number of cancer cells in a subject in need thereof relative to a control, comprising administering to the subject (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonist; and (c) one or more immune checkpoint inhibitors.

[0010] The present technology comprises a method of preventing or reducing a tumor volume in a subject in need thereof relative to a control, comprising administering to the subject (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonist; and (c) one or more immune checkpoint inhibitors.

[0011] The present technology comprises a method of preventing or reducing a cancer metastasis in a subject in need thereof relative to a control, comprising administering to the subject (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonist; and (c) one or more immune checkpoint inhibitors.

[0012] The present technology comprises a method of preventing or reducing a cancer in a subject in need thereof relative to a control, comprising administering to the subject (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonist comprising or consisting of bosentan or a derivative thereof; and (c) one or more immune checkpoint inhibitors.

[0013] The present technology comprises a method of reducing a number of cancer cells in a subject in need thereof relative to a control, comprising administering to the subject (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonist comprising or consisting of bosentan or a derivative thereof; and (c) one or more immune checkpoint inhibitors.

[0014] The present technology comprises a method of preventing or reducing a tumor volume in a subject in need thereof relative to a control, comprising administering to the subject (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonist comprising or consisting of bosentan or a derivative thereof; and (c) one or more immune checkpoint inhibitors.

[0015] The present technology comprises a method of preventing or reducing a cancer metastasis in a subject in need thereof relative to a control, comprising administering to the subject (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonist comprising or consisting of bosentan or a derivative thereof; and (c) one or more immune checkpoint inhibitors.

[0016] In some embodiments, the T cell composition comprises one or more of a CD3+T cell population, a CD2+T cell population, or a CD28+T cell population.

[0017] In some embodiments, the T cell composition comprises or consists of a (i) a CD3+T cell population, (ii) a CD3+CD28+T cell population, (iii) a CD3+CD2+T cell population, or (iv) a CD3+CD2+CD28+T cell population.

[0018] In some embodiments, the T cell population comprises one or more T cells.

[0019] In some embodiments, the method reduces a number of cancer cells in the subject, relative to the control.

[0020] In some embodiments, the number of cancer cells comprises an increase in a cancer cell size or an increase in a cancer cell number, relative to baseline.

[0021] In some embodiments, the method prevents or reduces a tumor volume in the subject, relative to the control.

[0022] In some embodiments, the method prevents or reduces a cancer metastasis in the subject, relative to the control.

[0023] In some embodiments, the tumor or cancer comprises a solid tumor.

[0024] In some embodiments, the cancer comprises a metastatic cancer.

[0025] In some embodiments, the cancer is a breast cancer.

[0026] In some embodiments, the breast cancer is a HER2 positive breast cancer.

[0027] In some embodiments, the breast cancer is a HER2 negative breast cancer.

[0028] In some embodiments, the breast cancer is a HER2 / neu-negative breast cancer.

[0029] In some embodiments, the breast cancer is a triple negative breast cancer.

[0030] In some embodiments, the breast cancer is selected from the group consisting of a stage I breast cancer, a stage II breast cancer, a stage III breast cancer, and a stage IV breast cancer.

[0031] In some embodiments, preventing the cancer, the number of cancer cells, the tumor volume, or the cancer metastasis comprises preventing an advancement of a cancer stage or a cancer grade.

[0032] In some embodiments, the advancement of the cancer stage comprises a stage 0 cancer progressing to a stage I cancer, a stage I cancer progressing to a stage II cancer, a stage II cancer progressing to a stage III cancer, or a stage III cancer progressing to a stage IV cancer.

[0033] In some embodiments, the advancement of the cancer grade comprises a grade 1 cancer progressing to a grade 2 cancer, a grade 2 cancer progressing to a grade 3 cancer, or a grade 3 cancer progressing to a grade 4 cancer.

[0034] In some embodiments, the number of cancer cells is reduced by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100%, relative to a control.

[0035] In some embodiments, the tumor volume is reduced by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100%, relative to a control.

[0036] In some embodiments, the cancer metastasis is reduced by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100%, relative to a control.

[0037] In some embodiments, the reduction or prevention in metastasis comprises a reduction in one or more of a number of metastasis, a size of metastasis, a rate of metastasis growth, a recurrence of metastasis after a cancer therapy.

[0038] In some embodiments, the T cell composition is administered at a dose comprising at least about of 5 x 104T cells to at least about 15 x 1010cells.

[0039] In some embodiments, the method further comprises administering one or more immune checkpoint inhibitors to the subject.

[0040] In some embodiments, the one or more immune checkpoint inhibitors is selected from the group consisting of a CTLA-4 inhibitor, a PD-1 inhibitor, a PD-L1 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor, a TIG IT inhibitor, a VISTA inhibitor, a VEGFR1 inhibitor, a VEGFR2 inhibitor, and a VEGFR3 inhibitor.

[0041] In some embodiments, the one or more immune checkpoint inhibitors comprises an antibody.

[0042] In some embodiments, the antibody is selected from the group consisting of pembrolizumab, ipilimumab, nivolumab, atezolizumab, cemiplimab, avelumab, durvalumab, and tremelimumab.

[0043] In some embodiments, the one or more immune checkpoint inhibitors are administered at a dose of about 0.5 mg / kg to about 10 mg / kg.

[0044] In some embodiments, the endothelin receptor antagonist comprises bosentan or a derivative thereof.

[0045] In some embodiments, the derivative of bosentan comprises a bosentan salt.

[0046] In some embodiments, the endothelin receptor antagonist is administered at a dose of about 0.5 mg to about 600 mg.

[0047] In some embodiments, a first dose of the T cell composition is administered to the subject prior to administration of a first dose the one or more immune checkpoint inhibitors.

[0048] In some embodiments, a first dose of the one or more immune checkpoint inhibitors is administered to the subject prior to a first dose of the T cell composition.

[0049] In some embodiments, the subject is administered at least about three doses one or more immune checkpoint inhibitors.

[0050] In some embodiments, the three or more doses of the one or more immune checkpoint inhibitors are administered to the subject about every three weeks to about every four weeks.

[0051] In some embodiments, a first dose of the endothelin receptor antagonist is administered to the subject prior to administration of a first dose the T cell composition.

[0052] In some embodiments, a second dose of the endothelin receptor antagonist is administered to the subject prior to administration of the first dose of the T cell composition.

[0053] In some embodiments, the subject is administered the endothelin receptor antagonist daily.

[0054] In some embodiments, the one or more cancer antigens are selected from the group consisting of MAGE-A4, MAGEA1 , NY-ESO-1 , KK-LC-1 , FRAME, B7-H3, B7-H4, CD19, and CD22.

[0055] In some embodiments, the T cell composition has an increase in one or more gene expression levels selected from the group consisting of a perforin-1 (PRF1 ) gene expression level, a granzyme B (GZMB) gene expression level, an interferon gamma (INFy) gene expression level, a tumor necrosis factor alpha (TNFa) gene expression level, a human leukocyte antigen class II histocompatibility antigen, DR alpha chain (HLA-DRA) gene expression level, a CD25 gene expression level, a CD69 gene expression level, and an IL- 2 gene expression level, relative to a control.

[0056] In some embodiments, the T cell composition has an increase in one or more protein levels selected from the group consisting of a PRF1 protein level, a GZMB protein level, an INFy protein level, a TNFa protein level, an HLD-DRA protein level, a CD25 protein level, a CD69 protein level, and an IL-2 protein level, relative to a control.

[0057] In some embodiments, at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100% of the T cells in the T cell composition are specific to the one or more cancer antigens.

[0058] In some embodiments, the T cell composition comprises T cells that are derived from a subject having a cancer.

[0059] In some embodiments, the T cells that are derived from a subject having cancer comprises T cells that were isolated or obtained from the subject having cancer.

[0060] In some embodiments, the cancer comprises a solid tumor.

[0061] In some embodiments, the cancer comprises a metastatic cancer.

[0062] In some embodiments, the cancer is a breast cancer.

[0063] In some embodiments, the breast cancer is a HER2 positive breast cancer.

[0064] In some embodiments, the breast cancer is a HER2 negative breast cancer.

[0065] In some embodiments, the breast cancer is a HER2 / neu-negative breast cancer.

[0066] In some embodiments, the breast cancer is a triple negative breast cancer.

[0067] In some embodiments, the breast cancer is selected from the group consisting of a stage I breast cancer, a stage II breast cancer, a stage III breast cancer, and a stage IV breast cancer.

[0068] In some embodiments, one or more of the T cell compositions, the endothelin receptor antagonist, or the immune checkpoint inhibitor are formulated for subcutaneous, intravenous, intramuscular, intracranial, intra-arterial, intraventricular, intranodal, intratumor, or intraperitoneal administration.

[0069] In some embodiments, the T cell compositions is formulated for subcutaneous, intravenous, intramuscular, intracranial, intra-arterial, intraventricular, intranodal, intratumor, or intraperitoneal administration.

[0070] In some embodiments, one or more of the T cell compositions, the endothelin receptor antagonist, or the immune checkpoint inhibitor are formulated in a vaccine.

[0071] In some embodiments, one or more of the T cell compositions, the endothelin receptor antagonist, or the immune checkpoint inhibitor are formulated in a nanoparticle.

[0072] In some embodiments, the endothelin receptor antagonist or the immune checkpoint inhibitor is formulated for oral administration or intravenous administration.

[0073] In some embodiments, the control comprises the subject at baseline.

[0074] In some embodiments, the control comprises a subject that is not administered one or more of (a) T cell composition specific to one or more cancer antigens; (b) an endothelin receptor antagonist; and (c) one or more immune checkpoint inhibitors.

[0075] In some embodiments, the control comprises a subject that is administered (a) alone.

[0076] In some embodiments, the control comprises a subject that is administered (b) alone.

[0077] In some embodiments, the control comprises a subject that is administered (c) alone.

[0078] In some embodiments, the control comprises a subject administered (a) and (b), but not (c).

[0079] In some embodiments, the control comprises a subject administered (b) and (c), but not (a).

[0080] In some embodiments, the control comprises a subject administered (a) and (c), but not (b).

[0081] In some embodiments, the endothelin receptor antagonist comprises bosentan.BRIEF DESCRIPTION OF THE DRAWINGS

[0082] FIG.1 illustrates an exemplary dosing scheme of an endothelin receptor antagonist (BOS), a T cell composition (ACT), and an immune checkpoint inhibitor (ICB) in mice, in accordance with the methods of the present technology. TDLNs: tumor-draining lymph nodes.

[0083] FIG. 2 illustrates changes in tumor volume in mice inoculated with 4T1 mammary carcinoma tumor that were administered an endothelin receptor antagonist (Bosentan), a T cell composition (ACT), an immune checkpoint inhibitor (ICB), or a combination thereof, in accordance with the methods of the present technology.

[0084] FIGS. 3A-3I illustrate the change in tumor volume of the individual mice of FIG. 2.

[0085] FIG. 4 illustrates a percent of response for the mice of FIGS. 3A-3I. PD: Progressive Disease; SD: Stable Disease; PR: Partial Response; CR: Complete Response; MCR: Maintained Complete Response.

[0086] FIG. 5 illustrates survival rates in the mice of FIG. 2 (N= 8 groups, 12 mice per group).

[0087] FIG. 6 illustrates lung metastasis in untreated 4T1 -tumor bearing mice compared to non-tumor bearing control mice. Arrows indicate macrometastatic lesions on the lung ventricles.

[0088] FIGS. 7A-7C illustrate tumor volume (FIGS. 7A and 7B) and survival fraction (FIG. 70) in mice administered an endothelin receptor antagonist (Bosentan), a T cell composition (ACT), an immune checkpoint inhibitor (ICB), or a combination thereof.

[0089] FIGS. 8A-8O illustrate survival fraction (FIG. 8A), change in tumor volume (FIG. 8B), and tumor mass following tumor section on day 21 (FIG. 80) in 4T1 -tumor bearing mice administered ACT alone or different combinations of ACT, Bosentan, and immune checkpoint inhibitors (ICIs) compared to control mice. In FIG. 8B, * 0.035, ** <0.007, *** 0.0003, **** <0.0001.

[0090] FIGs. 9A-9D illustrate tumor dynamics per mouse administered ACT-Bosentan versus control mice (FIG. 9A), tumor dynamics per mouse administered ACT-Bosentan-ICIs versus control mice (FIG. 9B), and average tumor size per group in mice administered ACT- Bosentan or ACT-Bosentan-ICIs versus control mice (FIG. 9C-9D) in a Renca mouse model of tumor-bearing mice.

[0091] FIG. 10 illustrates tumor volume on day 35 in the treatment groups of FIG. 9.

[0092] FIG. 11 illustrates the percent of responders and type on day 35 in the treatment groups of FIG. 9, where PD: Progressive Disease; SD: Stable Disease; PR: Partial Response; CR: Complete Response; MCR: Maintained Complete Response.

[0093] FIG. 12 illustrates survival rates of the treatment groups of FIG. 9.DETAILED DESCRIPTION

[0094] The present technology comprises T cell compositions and uses thereof for preventing or reducing cancer in subjects. The methods may comprise administering the T cell compositions, an endothelin receptor antagonist, and / or an immune checkpoint inhibitor to the subject.T Cell Compositions

[0095] The present technology comprises T cell compositions comprising one or more CD3+, CD2+, or CD28+donor-derived T cells. In some embodiments, the donor-derived T cells comprise a (i) CD3+T cell, (ii) CD3+CD28+T cell, (iii) CD3+CD2+ T cell, or (iv) CD3+CD2+CD28+T cell. The donor-derived T cells may comprise T cells isolated from peripheralblood (e.g., whole blood isolate or a peripheral blood mononuclear cell isolation). The T cells may be isolated using any means known in the art, including, but not limited to magnetic columns and / or beads (e.g., negative or positive selection) or cell sorting (e.g., fluorescence- activated cell sorting).

[0096] The T cell compositions are specific to at least a portion of one or more cancer antigens. The T cell compositions may comprise one or more T cells that bind to and / or recognize at least a portion of one or more cancer antigens. The portion of the one or more cancer antigens may comprise a protein, a peptide, or an amino acid motif that is associated with a cancer cell.

[0097] In some embodiments, the T cell compositions comprise one or more T cells having one or more receptors that are specific to at least the portion of the one or more cancer antigens. Nonlimiting examples of the one or more cancer antigens include Melanoma-associated antigen 4 (MAGEA4; National Center for Biotechnology Information (NCBI) Accession: NP_001373125.1 ), Melanoma-associated antigen 1 (MAGEA1 ; NCBI Accession: NP_004979.3); New York esophageal squamous cell carcinoma 1 (NY-ESO-1 ; NCBI Accession: CAA05908.1 ), Kita-Kyushu lung cancer antigen-1 (KK-LC-1 ; NCBI Accession: NP_001017978.1 ), Preferentially Expressed Antigen in Melanoma (PRAME; NCBI Accession: CAG30435.1 ), B7-H3 (NCBI Accession: Q5ZPR3.1 ); B7-H4 (NCBI Accession(s): AAZ17406.1 ; Q7Z7D3.1 ), Cluster of Differentiation 19 (CD19; NCBI Accession(s): AAB60697.1 ; AAA69966.1 ), Cluster of Differentiation 22 (CD22; NCBI Accession(s): NP_001762.2; NP_001172028.1 ; NP_001 172029.1 ; NP_001 172030.1 ; NP_001265346.1 ), KRAS protein (NCBI Accession(s): NP_001356715; NP_001356716) including its various isoforms and mutated forms (i.e., KRAS G12D), Fibroblast Growth Factor Receptor 3 (NCBI Accession(s): XP_054205236) including its various isoforms and mutated forms (i.e., FGFR3 Y373C), Melanoma Antigen Recognized by T cells (MART-1 ) (NCBI Accession(s): NP_005502), and Glycoprotein 100 (gp100) (NCBI Accession(s): AAC60634).

[0098] In some embodiments, about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100% of the T cells in the T cell composition are specific to the one or more cancer antigens.

[0099] In some embodiments, at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100% of the T cells in the T cell composition are specific to the one or more cancer antigens.

[0100] In some embodiments, at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100% of the T cells in the T cell composition are specific to the one or more cancer antigens.

[0101] The T cell compositions may comprise an increase in one or more gene expression levels relative to a control. In some embodiments, the T cell compositions comprise an increase in one or more gene expression levels selected from the group consisting of a perforin-1 (PRF1 ; NCBI Gene ID: 5551 ) gene expression level, a granzyme B (GZMB; NCBI Gene ID: 3002) gene expression level, an interferon gamma (INFy; NCBI Gene ID: 3458) gene expression level, a tumor necrosis factor (TNF; NCBI Gene ID: 7124) gene expression level, a human leukocyte antigen class II histocompatibility antigen, DR alpha chain (HLA-DRA; NCBI Gene ID: 3122) gene expression level, a CD25 (NCBI Gene ID: 3559) gene expression level, a CD69 level (NCBI Gene ID: 969) gene expression, and an interleukin-2 (IL-2; NCBI Gene ID: 3558) gene expression level, relative to a control.

[0102] The T cell compositions may comprise an increase in one or more protein levels, relative to a control. In some embodiments, the T cell compositions comprise an increase in one or more protein levels selected from the group consisting of a PRF1 protein level (NCBI Accession(s): KAI4076276.1 ; KAI4076275.1 ), a GZMB protein level (NCBI Accession(s): P10144.2; NP_001332940.1 ; NP_004122.2; EAW66003.1 ; EAW66002.1 ; AAH30195.1 ), an INFy protein level (NCBI Accession(s): AAB59534.1 ; AAA53230.1 ; AAM28885.1 ), a TNFa protein level, (NCBI Accession(s): CAA78745.1 ; BAF31279.1 ; P01375.1 ; NP_000585.2), an HLA-DRA protein level (NCBI Accession(s): UQL51 198.1 ; NP_061984.2), a CD25 protein level (NCBI Accession(s): NP_000408.1 ; NP_001295171 .1 ; NP_001295172.1 ), a CD69 protein level (NCBI Accession(s): AAH20057.1 ; NP_391988.1 ; EAW85959.1 ; EAW85956.1 ; EAW85952.1 ; EAW85951.1 ; EAW85949.1 ), and an IL-2 protein level (NCBI Accession(s): P60568.1 ; EAX05227.1 ; NP_000577.2; AAA70092.1 ; AAA59140.1 ), relative to a control.

[0103] The T cell composition may comprise T cells that are derived from a subject having a cancer. The T cells that are derived from the subject having a cancer may comprise T cells that were isolated or obtained from the subject.

[0104] The cancer may comprise a solid tumor and / or a metastatic cancer. In some embodiments, the cancer is a breast cancer (e.g., a HER2 positive breast cancer, a HER2 negative breast cancer, a HER2 / neu-negative breast cancer, or a triple negative breast cancer) or a B cell malignancy. The breast cancer may be selected from the group consisting of a stage I breast cancer, a stage II breast cancer, a stage III breast cancer, and a stage IV breast cancer. The B cell malignancy may comprise leukemia (e.g., B-cell acute lymphoblastic leukemia) or lymphoma.Methods of Generating T Cell Compositions

[0105] The T cell compositions of the present technology may be generated by one or more steps of (a) isolating a T cell population from a donor; (b) culturing the T cell population with one or more cytokines; (c) culturing the T cell population with one or more antibodies; and (d) expanding the T cell population for a period of time sufficient to generate the T cell composition.

[0106] The T cell population in may comprise an autologous T cell population or an allogenic T cell population. In some embodiments, isolating the T cell population in (a) comprises obtaining a T cell population. The isolating or obtaining the T cell population may comprise purifying or selecting for a single cell suspension. The single cell suspension may be purified from a tumor or a tumor-draining lymph node.

[0107] In some embodiments, the isolating or obtaining of the T cell composition comprises isolating or obtaining T cells from peripheral blood (e.g., whole blood isolate or a peripheral blood mononuclear cell isolation). The T cells may be isolated using any means known in the art, including, but not limited to magnetic columns and / or beads (e.g., negative or positive selection) or cell sorting (e.g., fluorescence-activated cell sorting (FACS)).

[0108] In some embodiments, the one or more cytokines in (b) may comprise or consist of IL-2. In some embodiments, the one or more cytokines in (b) comprise or consist of IL-2, interleukin 7 (IL-7), interleukin 15 (IL-15), interleukin 4 (IL-4), interleukin 12 (IL-12), andinterleukin 18 (IL-18), or any combination thereof. In some embodiments, the culturing the T cell population with one or more cytokines in (b) is an optional step.

[0109] The one or more antibodies in (c) may comprise or consist of (i) an anti-CD3 antibody, (ii) an anti-CD3 antibody and an anti-CD28 antibody, or (iii) an anti-CD3 antibody and an anti-CD2 antibody. In some embodiments, step (c) occurs simultaneously with step (b). In some embodiments, step (c) is optional.

[0110] In some embodiments, the period of time in (d) comprises about 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 12 days, 14 days, 16 days, 18 days, 20 days, 22 days, 24 days, 28 days, 30 days, 35 days, or 40 days.

[0111] In some embodiments, the period of time in (d) comprises at least 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 12 days, 14 days, 16 days,18 days, 20 days, 22 days, 24 days, 28 days, 30 days, 35 days, or 40 days.

[0112] In some embodiments, the period of time in (d) comprises at least about 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 12 days, 14 days, 16 days,18 days, 20 days, 22 days, 24 days, 28 days, 30 days, 35 days, or 40 days.

[0113] Step (d) may occur simultaneously to step (c), or step (d) may be optional.

[0114] In some embodiments, the T cell population in (c) or (d) comprises one or more pre-rapid expansion protocol (pre-REP) T cells. The pre-REP T cells may comprise a tumorinfiltrating lymphocyte (TIL) that has been dissociated from a tumor fragment.

[0115] In some embodiments, (d) comprises one or more rapid expansion protocols (REP). The REP may comprise stimulating with one or more T cells in the T cell population with one or more costimulatory agents. Nonlimiting examples of costimulatory agents include a cytokine (e.g., IL-2), an antibody (e.g., an anti-CD3 antibody, an anti-CD28 antibody, an anti-CD2 antibody), a ligand or cancer antigen (e.g., CD40 ligand (CD40L) (NCBI Accession: NP_000065.1 ), 4-1 BB (NCBI Accession: AAA53133.1 ), or 0X40 (NCBI Accession: CAE1 1757.1 ), or one or more irradiated feeder cells.

[0116] One or more of steps (a)-(d) may comprise expanding the T cell population according to any means known in the art. In some embodiments, one or more of steps (a)-(d) are conducted according to the methods disclosed in Zhao Y et al., High-efficiency transfection of primary human and mouse T lymphocytes using RNA electroporation. Mol Ther. 2006 Jan;13(1 ) :151 -9 or Kverneland AH, et al., Adoptive cell therapy in combination with checkpoint inhibitors in ovarian cancer. Oncotarget. 2020 Jun 2;1 1 (22):2092-2105, each incorporated herein by reference in their entireties.Engineering T cells

[0117] The T cells in the T cell compositions of the present technology may be engineered to increase or decrease expression of one or more genes and / or levels of one or more proteins, relative to baseline.

[0118] In some embodiments, one or more T cells in the T cell compositions are engineered to express an exogenous T cell receptor or increase expression of an endogenous T cell receptor, relative to baseline. In some embodiments, the T cell receptor is a receptor specific to a cancer antigen selected from the group consisting of MAGE-A4, MAGEA1 , NY-ESO-1 , KK-LC-1 , FRAME, B7-H3, B7-H4, CD19, and CD22. In some embodiments, the T cell receptor comprises a chimeric antigen receptor (CAR) and / or a CC- chemokine receptor (COR). In some embodiments, the T cell receptor is specific to a cancer antigen disclosed in Xiao, J. et al., Expression of four cancer-testis antigens in TNBC indicating potential universal immunotherapeutic targets. J Cancer Res Clin Oncol 149, 15003-1501 1 (2023) or Wescott, Elizabeth C et al., Epithelial Expressed B7-H4 Drives Differential Immunotherapy Response in Murine and Human Breast Cancer. Cancer research communications vol. 4,4 (2024): 1 120-1134, each incorporated herein by reference in their entireties.

[0119] In some embodiments, one or more T cells in the T cell compositions are engineered to express an exogenous cytokine or to increase expression of an endogenous cytokine, relative to baseline. The cytokines may comprise any of those disclosed in Zhang L et al., Improving adoptive T cell therapy by targeting and controlling IL-12 expression to the tumor environment. Mol Ther. 201 1 Apr;19(4):751 -9 or in Stephan MT et al., T herapeutic cell engineering with surface-conjugated synthetic nanoparticles. Nat Med. 2010 Sep;16(9):1035-41 . doi: 10.1038 / nm.2198, each incorporated herein by reference in their entireties.

[0120] One or more T cells in the T cell compositions may be engineered to reduce or remove expression of an inhibitory receptor (e.g., PD-1 , TIM-3, LAG-3, TIGIT) or a transcript of a gene encoding an inhibitory receptor. In some embodiments, The T cells in the T cell compositions are engineered to reduce or remove expression of T-cell receptor a constant (TRAC) transcript or a TRAC protein level, relative to baseline.

[0121] In some embodiments, the T cell compositions specifically bind and / or recognize the cancer antigen by a T cell receptor, a chimeric antigen receptor (CAR), and / or a CC-chemokine receptor (CCR).

[0122] Exemplary CARS'. The CARs may comprise one or more of an antigen recognition domain, a hinge region, a transmembrane domain, and an endodomain (e.g., an intracellular signaling domain), each of which may be derived from one or more sources, including naturally occurring and synthetic sources. In some embodiments, the CARs are donor-derived and / or cord blood derived. In some embodiments, the CAR is a humanized CAR.

[0123] The antigen recognition domain may bind and / or specifically recognize a target antigen (e.g., a cancer antigen). The antigen recognition domain may be an extracellular antigen recognition domain. The antigen recognition domain may comprise a single-chain variable fragment (scFv) derived from one or more antibodies or antigen binding fragments thereof. In some embodiments, the antigen recognition domain may comprise or consist of at least a portion of a nanobody, a ligand (e.g., a natural ligand or a receptor portion thereof, such as NKG2D, or a PD-1 extracellular domain), or an alternative binding protein (i.e., a protein which acts as a binding agent but is not a traditional antibody). In some embodiments, the antigen recognition domain may be an anti-CD19 antibody, such as an human anti-CD19 scFv. In some embodiments, the antigen recognition domain may be an FMC63 monoclonal antibody that binds human CD19

[0124] The hinge region may provide flexibility and / or enhance spatial arrangement between the antigen recognition domain and the transmembrane domain, relative to a control. The hinge region may be at least partially derived from an IgG, a CD8 peptide, or a CD28 peptide. The hinge region may also enhance binding of the CAR to a target antigen, relative to a control.

[0125] The transmembrane domain may promote or enhance anchoring of the CAR to the membrane of one or more T cells in the T cell composition. The transmembrane domain may be derived from a transmembrane region of an immunological protein, including those selected from the group consisting of CD3^, CD8, CD28, and CD4.

[0126] The endodomain may comprise or consist of an intracellular signaling domain. In some embodiments, the CARs comprise two or more endodomains. The endodomain may promote or stimulate one or more T cells in the T cell composition during or after cancer antigen binding. The endodomain may comprise, consist of, or be derived from an intracellular signaling domain from CD3^ (zeta chain), which may act as part of the T cell receptor complex and delivers the activation signal required for T cell function. The endodomain may also comprise one or more co-stimulatory signaling domains (such as CD2, CD28, 4-1 BB [CD137], 0X40, or ICOS) (e.g., in addition to CD3Q. The co-stimulatory domains may enhance an activation, survival, proliferation, and / or cytotoxic function of the T cell composition.

[0127] In some embodiments, the CAR specifically binds and / or recognizes CD19 (e.g., a CD19 antigen) or at least a portion thereof (e.g., two or more amino acids). In some embodiments, the CAR is an anti-CD19 CAR. In some embodiments, the CAR comprises or consists of a CAR of a therapy selected from the group consisting of tisagenlecleucel, axicabtagene ciloleucel, lisocabtagene maraleucel, brexucabtagene autoleucel, ciltacabtagene autoleucel, and idecabtagene vicleuce. In some embodiments, the CAR comprises or consists of a CAR of a therapy selected from the group consisting of (i) NXC- 201 , HBI0101 , IM19, CAR-GPRC5D, CDH17 / GUCY2C CAR-T, SL1904B, AIC100, SCT- 001 , GD2CAR, Senl-h19 CAR-T, LMY-920, SNC-109, t aPD1 -MSLN-CAR T Cells, CD19x22 CAR T, IL13Ra2 CAR-T, LMP1 CAR- T, IM96, FL-33, KD-025, GPC3 CAR, WL276, NKG2D CAR-T, Anti-CEA CAR-T, CD7 CAR-T, CD318-CAR, CT125B, CD19 / CD22 CAR-T, Claudin18.2-Targeted CAR, APRIL CAR-T, BCMA CAR-T, ADGRE2 CAR-T, CAIX-targeted CAR-T, EphA2-targeted CAR, CLL1 CAR, CD38 CAR, Anti-FcRL5 CAR, GPC3-CAR, CD70 CAR, CD123 CAR, Anti-CD56-CAR-T, Anti-CLL1 / +CD33 CAR T Cells, those of (ii) NCT06758713, NCT07143929, NCT06828341 , NCT06010862, NCT05618041 ,NCT06215950, NCT04572308, NCT04546906, NCT03258047, or those (iii) disclosed inKfir-Erenfeld S, et al, Feasibility of a Novel Academic BCMA-CART (HBI0101) for the Treatment of Relapsed and Refractory AL Amyloidosis. Clin Cancer Res. 2022 Dec 1 ;28(23):5156-5166; Asherie N, et al, Development and manufacture of novel locally produced anti-BCMA CAR T cells for the treatment of relapsed / refractory multiple myeloma: results from a phase I clinical trial. Haematologica. 2023 Jul 1 ;108(7) :1827-1839, each of which are incorporated herein by reference in their entireties.

[0128] Tumor-infiltrating lymphocytes (TILs): In some embodiments, the T cell compositions comprise or consist of TILs. In some embodiments, the T cell compositions comprise two or more TILs. The TILs may comprise immune cells, such as T cells, derived from a tumor or otherwise cancerous tissue. The TILs may be isolated from the tumor or cancerous tissue, selected for based on a cancer antigen recognition associated with the tumor or cancerous tissue, and may be optionally expanded (e.g., ex vivo expansion). The TILs may specifically bind and / or recognize the cancer antigen. Additionally, the TILs may be used to target a cancer of the present technology without genetically modifying the TILs. The TILs may be derived from a donor, such as an autologous donor. In some embodiments, the donor is the subject in need thereof of the present technology. In some embodiments, the TILs comprise LN-144 or LN-145.

[0129] T Cell Receptor (TCR)-transgenic T Cells: In some embodiments, the T cell compositions comprise of consist of TOR-transgenic T cells. In some embodiments the TCR transgenic T cells comprise two or more TCR-transgenic T cells. The TCR transgenic T cells may be engineered to express a cancer antigen-specific TCR that specifically bind to and / or recognizes the cancer antigen, such as when the cancer antigen is presented by a human leukocyte antigen (HLA) molecule. The TCR of the TCR-transgenic T cell may comprise a TCR that is a non-native TCR. In some embodiments, the TCR specifically binds to and / or recognizes an intracellular antigen associated with a tumor or otherwise cancerous tissue, which may be presented by the HLA molecule.

[0130] Tumor-Draining Lymph Nodes (TDLNs): In some embodiments, the T cell compositions are derived from tumor-draining lymph nodes (TDLNs) as TDLNs may contain cancer-specific T cells. In some embodiments, the T cell compositions are derived from a combination of TDLNs and tumor tissue to enrich for tumor-specific T cells.Associated Methods

[0131] The present technology comprises methods for preventing or reducing a cancer in a subject in need thereof relative to a control. In some embodiments, the present technology comprises methods of reducing a number of cancer cells in a subject in need thereof relative to a control. In some embodiments, the present technology comprises methods of preventing or reducing a tumor volume in a subject in need thereof relative to a control. In some embodiments, the present technology comprises methods of preventing or reducing cancer metastasis in a subject in need thereof relative to a control. In some embodiments, the methods increase a survival rate of the subject, relative to a control.

[0132] In some embodiments, the methods of the present technology (i) reduce a number of cancer cells in the subject (e.g., reducing or preventing an increase in a cancer cell size or an increase in a cancer cell number, relative to baseline); (ii) prevent or reduce a tumor volume in the subject, or (iii) prevent or reduce a cancer metastasis in the subject, relative to the control (e.g., a reduction in one or more of a number of metastasis, a size of metastasis, a rate of metastasis growth, a recurrence of metastasis after a cancer therapy).

[0133] In some embodiments, the methods of the present technology reduce a number of cancer cells by about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100%, relative to a control.

[0134] In some embodiments, the methods of the present technology reduce a number of cancer cells by at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100%, relative to a control.

[0135] In some embodiments, the methods of the present technology reduce a number of cancer cells by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100%, relative to a control.

[0136] In some embodiments, the methods of the present technology reduce a tumor volume by about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100%, relative to a control.

[0137] In some embodiments, the methods of the present technology reduce a tumor volume by at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100%, relative to a control.

[0138] In some embodiments, the methods of the present technology reduce a tumor volume by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100%, relative to a control.

[0139] In some embodiments, preventing the cancer, the number of cancer cells, the tumor volume, or the cancer metastasis comprises preventing an advancement of a cancer stage or a cancer grade. The advancement of the cancer stage may comprise a stage 0 cancer progressing to a stage I cancer, a stage I cancer progressing to a stage II cancer, a stage II cancer progressing to a stage III cancer, or a stage III cancer progressing to a stage IV cancer. In some embodiments, the advancement of the cancer grade comprises a grade 1 cancer progressing to a grade 2 cancer, a grade 2 cancer progressing to a grade 3 cancer, or a grade 3 cancer progressing to a grade 4 cancer.Steps of the Methods

[0140] The methods of the present technology may comprise or consist of administering (a) a T cell composition of the present technology, (b) an endothelin receptor antagonist, and (c) one or more immune checkpoint inhibitors. In some embodiments, the methods comprise or consist of administering (a) a T cell composition of the present technology and (b) an endothelin receptor antagonist. In some embodiments, the methods comprise or consist of administering a T cell composition of the present technology.

[0141] In some embodiments, the endothelin receptor antagonist comprises or consists of bosentan or a derivative thereof (e.g., a bosentan salt).

[0142] In some embodiments, the one or more immune checkpoint inhibitors is selected from the group consisting of a CTLA-4 inhibitor, a SLAMF6 inhibitor, a PD-1 inhibitor, a PD-L1 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor, a TIGIT inhibitor, a VISTA inhibitor, a VEGFR1 inhibitor, a VEGFR2 inhibitor, and a VEGFR3 inhibitor. In some embodiments, the one or more immune checkpoint inhibitors comprises an antibody (e.g.,pembrolizumab, ipilimumab, nivolumab, atezolizumab, cemiplimab, avelumab, durvalumab, and tremelimumab).

[0143] Uses of bosentan and a checkpoint inhibitor for solid tumors is disclosed in WIPO Publication Number WO 2022 / 125805 A1 of PCT / US2021 / 062663 which is incorporated herein by reference in its entirety.Subjects

[0144] The subjects of the present technology may have a cancer, may be suspected of having a cancer, or may be susceptible to developing a cancer. The cancer may comprise a solid tumor and / or a metastatic cancer. In some embodiments, the cancer is a breast cancer (e.g., a HER2 positive breast cancer, a HER2 negative breast cancer, a HER2 / neu- negative breast cancer, or a triple negative breast cancer) or a B cell malignancy. The breast cancer may be selected from the group consisting of a stage I breast cancer, a stage II breast cancer, a stage III breast cancer, and a stage IV breast cancer. The B cell malignancy may comprise leukemia (e.g., B-cell acute lymphoblastic leukemia) or lymphoma.

[0145] The subject may have a cancer stage selected from the group consisting of a stage 0 cancer, a stage I cancer, a stage II cancer, a stage III cancer, and a stage IV cancer. In some embodiments, the cancer comprises a grade 1 cancer, a grade 2 cancer, a grade 3 cancer, or a grade 4 cancer.

[0146] In some embodiments the subject is a human subject.Controls

[0147] The controls (i.e. , control subjects) of the present technology may comprise the subject at baseline or a subject that is not administered one or more of (a) T cell composition of the present technology; (b) an endothelin receptor antagonist of the present technology; or (c) one or more immune checkpoint inhibitors of the present technology. In some embodiments, the control comprises a subject that is administered: (a) alone; (b) alone; (c) alone; (a) and (b), but not (c); (b) and (c), but not (a); or (a) and (c), but not (b).

[0148] In some embodiments, the control comprises a subject that does receive the same (a) T cell composition of the present technology; (b) an endothelin receptor antagonistof the present technology; and / or (c) one or more immune checkpoint inhibitors of the present technology but receives them in a different order than the subject of the methods of the present technology.DosingT Cell Compositions

[0149] The T cell compositions of the present technology may be administered to the subject at a dose comprising at least about of 5 x 104T cells to at least about 15 x 1010cells.

[0150] In some embodiments, the T cell compositions are administered at a dose comprising about 5 x 104T cells, about 6 x 104T cells, about 7 x 104T cells, about 8 x 104T cells, about 9x 104T cells, about 10 x 104T cells, about 1 x 105T cells, about 5x 105T cells, about 10x 105T cells, about 1 x 106T cells, about 5x 106T cells, about 10x 106T cells, about 1 x 107T cells, about 5x 107T cells, about 10x 107T cells, about 1 x 108T cells, about 5x 108T cells, about 10x 108T cells, about 1 x 109T cells, about 5x 109T cells, about 10x 109T cells, about 1 x 1 O10T cells, about 2 x 1 O10T cells, about 3 x 1 O10T cells, about 4 x 1010T cells, about 5 x 1010T cells, about 6 x 1010T cells, about 7 x 1010T cells, about 8 x 1010T cells, about 9 x 1010T cells, about 10 x 1010T cells, about 11 x 1010T cells, about12 x 1010T cells, about 13 x 1010T cells, about 14 x 1010T cells, or about 15 x 1010T cells.

[0151] In some embodiments, the T cell compositions are administered at a dose comprising at least 5 x 104T cells, at least 6 x 104T cells, at least 7 x 104T cells, at least 8 x 104T cells, at least 9x 104T cells, at least 10 x 104T cells, at least 1 x 105T cells, at least 5x 105T cells, at least 10x 105T cells, at least 1 x 106T cells, at least 5x 106T cells, at least 10x 106T cells, at least 1 x 107T cells, at least 5x 107T cells, at least 10x 107T cells, at least 1 x 108T cells, at least 5x 108T cells, at least 10x 108T cells, at least 1 x 109T cells, at least 5x 109T cells, at least 10x 109T cells, at least 1 x 1010T cells, at least 2 x 1 O10T cells, at least 3 x 1 O10T cells, at least 4 x 1 O10T cells, at least 5 x 1 O10T cells, at least 6 x 1O10T cells, at least 7 x 1 O10T cells, at least 8 x 1 O10T cells, at least 9 x 1 O10T cells, at least 10 x 1010T cells, at least 1 1 x 1010T cells, at least 12 x 1010T cells, at least13 x 1010T cells, at least 14 x 1010T cells, or at least 15 x 1010T cells.

[0152] In some embodiments, the T cell compositions are administered at a dose comprising at least about 5 x 104T cells, at least about 6 x 104T cells, at least about 7 x 104T cells, at least about 8 x 104T cells, at least about 9x 104T cells, at least about 10 x 104T cells, at least about 1 x 105T cells, at least about 5x 105T cells, at least about 10x 105T cells, at least about 1 x 106T cells, at least about 5x 106T cells, at least about 10x 106T cells, at least about 1 x 107T cells, at least about 5x 107T cells, at least about 10x 107T cells, at least about 1 x 108T cells, at least about 5x 108T cells, at least about 10x 108T cells, at least about 1 x 109T cells, at least about 5x 109T cells, at least about 10x 109T cells, at least about 1 x 1010T cells, at least about 2 x 1010T cells, at least about 3 x 1010T cells, at least about 4 x 1010T cells, at least about 5 x 1010T cells, at least about 6 x 1010T cells, at least about 7 x 1010T cells, at least about 8 x 1010T cells, at least about 9 x 1010T cells, at least about 10 x 1 O10T cells, at least about 11 x 1 O10T cells, at least about 12 x 1010T cells, at least about 13 x 1010T cells, at least about 14 x 1010T cells, or at least about 15 x 101° T cells.

[0153] In some embodiments, a first dose of the T cell composition is administered to the subject prior to administration of a first dose the one or more immune checkpoint inhibitorsEndothelin Receptor Antagonist

[0154] The endothelin receptor antagonists of the present technology may be administered to the subject at a dose of at least about 0.5 mg to at least about 600 mg.

[0155] In some embodiments, the endothelin receptor antagonists are administered at a dose of about 0.5 mg , about 1 .0 mg, about 5.0 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 50 mg, about 75 mg, about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 200 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, or about 600 mg.

[0156] In some embodiments, the endothelin receptor antagonists are administered at a dose of at least 0.5 mg , at least 1 .0 mg, at least 5.0 mg, at least 10 mg, at least 15 mg, at least 20 mg, at least 25 mg, at least 50 mg, at least 75 mg, at least 100 mg, at least 150 mg,at least 200 mg, at least 250 mg, at least 200 mg, at least 350 mg, at least 400 mg, at least 450 mg, at least 500 mg, at least 550 mg, or at least 600 mg.

[0157] In some embodiments, the endothelin receptor antagonists are administered at a dose of at least about 0.5 mg , at least about 1 .0 mg, at least about 5.0 mg, at least about 10 mg, at least about 15 mg, at least about 20 mg, at least about 25 mg, at least about 50 mg, at least about 75 mg, at least about 100 mg, at least about 150 mg, at least about 200 mg, at least about 250 mg, at least about 200 mg, at least about 350 mg, at least about 400 mg, at least about 450 mg, at least about 500 mg, at least about 550 mg, or at least about 600 mg.

[0158] The endothelin receptor antagonist may be administered daily. In some embodiments, a first dose of the endothelin receptor antagonist is administered to the subject prior to administration of a first dose the T cell composition. In some embodiments, a second dose of the endothelin receptor antagonist is administered to the subject prior to administration of the first dose of the T cell composition.Immune Checkpoint Inhibitors

[0159] The immune checkpoint inhibitors of the present technology may be administered to the subject in at least about one or more doses, two or more doses, or three or more doses. In some embodiments, the three or more doses of the one or more immune checkpoint inhibitors are administered to the subject about every three to four weeks. In some embodiments, a first dose of one or more immune checkpoint inhibitors is administered to the subject prior to a first dose of the T cell composition.

[0160] In some embodiments, the immune checkpoint inhibitors are administered at a dose of about 0.5 mg / kg to about 10 mg / kg. In some embodiments, the immune checkpoint inhibitors are administered at a dose of about 0.5 mg / kg, about 0.6 mg / kg, about 0.7 mg / kg, about 0.8 mg / kg, about 0.9 mg / kg, or about 10 mg / kg.

[0161] In some embodiments, the immune checkpoint inhibitors are administered at a dose of at least 0.5 mg / kg to at least 10 mg / kg. In some embodiments, the immune checkpoint inhibitors are administered at a dose of at least 0.5 mg / kg, at least 0.6 mg / kg, at least 0.7 mg / kg, at least 0.8 mg / kg, at least 0.9 mg / kg, or at least 10 mg / kg.

[0162] In some embodiments, the immune checkpoint inhibitors are administered at a dose of at least about 0.5 mg / kg to at least about 10 mg / kg. In some embodiments, the immune checkpoint inhibitors are administered at a dose of at least about 0.5 mg / kg, at least about 0.6 mg / kg, at least about 0.7 mg / kg, at least about 0.8 mg / kg, at least about 0.9 mg / kg, or at least about 10 mg / kg.

[0163] The immune checkpoint inhibitors may be administered about 6 to about 12 months. In some embodiments, the immune checkpoint inhibitors are administered for about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, or about 12 months.

[0164] The immune checkpoint inhibitors may be administered at least 6 to at least 12 months. In some embodiments, the immune checkpoint inhibitors are administered for at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 1 1 months, or at least 12 months.

[0165] The immune checkpoint inhibitors may be administered at least about 6 to at least about 12 months. In some embodiments, the immune checkpoint inhibitors are administered for at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, or at least about 12 months.

[0166] In some embodiments, the immune checkpoint inhibitors are administered to the subject until the subject experiences relapse or resistance, or one or more symptoms thereof, to one or more immune checkpoint inhibitors.Formulations

[0167] The T cell compositions, the endothelin receptor antagonists, or the immune checkpoint inhibitors of the present technology may be formulated for administration to the subject by any means known in the art. In some embodiments, one or more of the T cell compositions, the endothelin receptor antagonists, or the immune checkpoint inhibitors are formulated for subcutaneous, intravenous, intramuscular, intracranial, intra-arterial, intraventricular, intranodal, intratumor, intraperitoneal, or oral administration.

[0168] The T cell compositions, the endothelin receptor antagonists, or the immune checkpoint inhibitors may be formulated in a vaccine or a nanoparticle.EXAMPLESExample 1 : Generation of T Cell Compositions

[0169] 4T1 Mammary Carcinoma Model: T cell compositions of the present technology were generated and assessed in a murine cancer model (FIG. 1 ). The T cell compositions (ACT) were generated from a 4T1 mammary carcinoma model (4T1 -ACT). For the generation of the 4T1 -ACT model, female BALB / c mice (7 to 8 weeks old) were subcutaneously injected with 5 x 1044T1 cells and allowed to develop tumors until they reached —100 mm3in size. 10-13 days post-tumor engraftment tumor-bearing mice (Donors) were sacrificed, and their tumor-draining lymph nodes (TDLNs) were harvested. For the adoptive cellular therapy (ACT), T cells (Tc) were isolated from TDLNs, by a negative selection process of a TDLN single cell suspension. T cells were activated in vitro using activating antibodies to either CD3 or CD3 in combination to CD28 or CD2 for 48-72 hours in the presence of IL-2 (50IU / ml). Following removal of stimulation, T cells where further expanded in the presence of IL-2 for an additional 4 days. T cell activation was assessed via flow cytometric analysis of CD69 and CD25 expression at 48-72hrs post activation. Expanded T cells were phenotyped by flow cytometry to determine the % of T cell subsets.

[0170] Renca Tumor Model: T cell compositions (ACT) were generated from a Renca renal adenocarcinoma model using a method similar to that described for 4T 1 model above. For the generation of the Renca-ACT model, BALB / c mice (7 to 8 weeks old) were subcutaneously injected with 4 x 105Renca cells and allowed to develop tumors until they reached -100 mm3in size. 28 days post-tumor engraftment tumor-bearing mice (Donors) were sacrificed, and their tumor-draining lymph nodes (TDLNs) were harvested. For the adoptive cellular therapy (ACT), T cells (Tc) were isolated from TDLNs, by a negative selection process of a TDLN single cell suspension. T cells were activated in vitro using activating antibodies to either CD3 or CD3 in combination to CD28 or CD2 for 48-72 hours. Following removal of stimulation, T cells where further expanded in the presence of IL-2 (50IU / ml). for an additional 4 days. T cell activation was assessed via flow cytometricanalysis of OD69 and CD25 expression at 48-72hrs post activation. Expanded T cells were phenotyped by flow cytometry to determine the % of T cell subsets.Example 2: Impact of T Cell Compositions on Tumor Growth and Survival (4T1 Tumor Model)Assessing Tumor Size

[0171] For the in vivo 4T1 -ACT study, T cell compositions were intravenously infused into 4T1 tumor-bearing recipient mice (1 x 106T-cells / mouse) and tumor growth was monitored throughout the experiment; mice were sacrificed when tumor size reached humane endpoint (1200mm3). Tumor lung metastasis was also monitored when mice were sacrificed at day 28 or humane endpoint.Assessing Efficacy of T Cell Compositions, Immune Checkpoint Inhibitors, and / or Endothelin Receptor Antagonists Combination Therapies

[0172] Tumor Volume: T o improve the efficacy of the ACT, 4T 1 -mice were treated with ACT as a monotherapy or in combination with immune checkpoint inhibitors (ICB) and / or Bosentan (BOS), an endothelin receptor antagonist, and their corresponding single treatment controls (FIG. 2, FIGS. 3A-3I, and FIG. 4). For combination treatments, prior to infusion with T cell compositions between days 10 to 1 1 , recipient mice were treated intraperitoneally with a single dose of BOS 2 to 3 days before and continued receiving daily BOS treatment until day 21. Additionally, three doses of ICBs — 10 mg / kg anti-PD1 and 5 mg / kg anti-CTLA-4 — were administered starting 72 hours post-Tc infusion and repeated on days 17, and 21. The corresponding single treatment controls followed the same dosages and timings. Statistical significance in tumour volume growth between groups was determined with a t test at day 28 (FIG. 3I). Tumor growth was monitored throughout the study and compared to controls (FIG. 6).

[0173] Three mice from the triple combination group (T cell composition, ICB, and Bosentan-treated mice) maintained complete response as shown by complete regression of the primary tumors. These mice were rechallenged with a second 4T1 tumor cell line inoculation (5 x 1044T1 cells) and the tumor growth was monitored (FIGS. 7A and 7B). Thiswas done in parallel to 4T 1 tumor cell line inoculation of 10 untreated, tumor-inexperienced, aged-, gender- matched mice (control).

[0174] Survival: Mouse survival was monitored further up to day 109 (FIG. 5). Survival was assessed on 8 groups with 12 mice per group. In each of the ACT-ICB and ACT- Bosentan groups, 2 mice died by day 21 despite having a low tumor volume, similar to 1 mouse in the ACT alone group and 3 mice in the ACT-ICB-Bosentan treated group. These mice were excluded from the survival study to better-represent deaths due to tumor or tumor metastasis.

[0175] The surviving mice were re-challenged with 5 x 1044T1 cells and the tumor growth and survival were monitored throughout the duration of the experiment over a period of 33 days of tumor inoculation. This was done in parallel to 4T1 tumor cell line primary inoculation (5 x 1044T1 cells) of tumor-inexperienced, untreated aged-, gender-matched mice (control) (FIG. 7C). Mice were sacrificed at day of termination of experiment or when tumors reached a humane endpoint of 1200mm3.

[0176] Removal of Tumors (Day 21)\ Tumor volume and survival experiments were performed as described above with the following deviation: upon completion of treatment regimens, the tumors from all groups were surgically removed on day 21 . From then on, a survival study was performed for the remaining mice that survived the surgery. FIG. 8A shows mouse survival fractions until day 1 16 post tumor inoculation. As can be seen, the triple therapy (Bosentan-ICIs-ACT) appears to significantly inhibit 4T1 metastasis that ultimately results in animal death. FIG. 8B shows the average tumor size in the different treatment groups over 22 days. The triple combination therapy (Bosentan-ICIs-ACT) provides more robust anti-tumor effects compared with ACT monotherapy and the Bosentan-ICIs, Bosentan-ACT, and ICIs-ACT dual therapies. Tumor mass in each treatment group following tumor section on day 21 is shown in FIG. 8C.Example 3: Impact of T Cell Compositions on Tumor Growth and Survival (Renca Tumor Model)

[0177] For the in vivo Renca-ACT study, T cell compositions were intravenously infused into Renca tumor-bearing recipient mice (1 x 106T-cells / mouse) and tumor growthwas monitored throughout the experiment. Mice were sacrificed when tumor size reached humane endpoint (1200mm3).

[0178] Assessing Efficacy of T Cell Compositions, Immune Checkpoint Inhibitors, and / or Endothelin Receptor Antagonist Combination Therapies

[0179] Tumor Volume: Renca-mice were treated with a combination of ACT and Bosentan with or without ICIs. Bosentan treatment was started on day 20 and then daily until day 28, inclusive. ACT was given on day 21 , and ICI treatment was given on days 22, 25, and 28. FIGs. 9A-9B show changes in tumor volume in individual mice, where 6 mice were in the control group, 5 mice were in the ACT-Bosentan treatment group, and 6 mice were in the ACT-Bosentan-ICIs treatment group. FIGs. 9C-9D show the average tumor volume per treatment group. FIG. 10 shows tumor volume in each treatment group on day 35.

[0180] The response to each treatment combination was assessed using a five category method where CR = complete response, MCR = maintained complete response, PR = partial response, SD = stable disease, and PD = progressive disease. For CR, complete tumor regression in at least one assessment with regrowth was observed before the end of the experiment. For MCR (Maintained Complete Response), mice remained tumor free until the end of the experiment. The percent response was quantified in each treatment group at the last time point of the experiment (day 36) (FIG. 1 1 ). As can be seen, the combination of ACT and Bosentan both with and without ICIs slows down Renca kidney cancer growth.

[0181] Survival: Mouse survival was monitored up to the last time point (day 36). As shown in FIG. 12, the combination of ACT and Bosentan both with and without ICIs extends overall survival in Renca-tumor bearing mice compared with untreated control mice.Additional Embodiments

[0182] Various embodiments of the present technology are set forth below in paragraphs

[0183] to

[0338] :

[0183] 1 . A method of preventing or reducing a cancer in a subject in need thereof relative to a control, comprising administering to the subject (a) a T cell composition specificto at least a portion of one or more cancer antigens; and (b) an endothelin receptor antagonist.

[0184] 2. A method of reducing a number of cancer cells in a subject in need thereof relative to a control, comprising administering to the subject (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; and (b) an endothelin receptor antagonist.

[0185] 3. A method of preventing or reducing a tumor volume in a subject in need thereof relative to a control, comprising administering to the subject (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; and (b) an endothelin receptor antagonist.

[0186] 4. A method of preventing or reducing cancer metastasis in a subject in need thereof relative to a control, comprising administering to the subject (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; and (b) an endothelin receptor antagonist.

[0187] 5. A method of preventing or reducing a cancer in a subject in need thereof relative to a control, comprising administering to the subject (a) a T cell composition specific to one or more cancer antigens; (b) an endothelin receptor antagonist; and (c) one or more immune checkpoint inhibitors.

[0188] 6. A method of preventing or reducing a number of cancer cells in a subject in need thereof relative to a control, comprising administering to the subject (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonist; and (c) one or more immune checkpoint inhibitors.

[0189] 7. A method of preventing or reducing a tumor volume in a subject in need thereof relative to a control, comprising administering to the subject (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonist; and (c) one or more immune checkpoint inhibitors.

[0190] 8. A method of preventing or reducing a cancer metastasis in a subject in need thereof relative to a control, comprising administering to the subject (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonist; and (c) one or more immune checkpoint inhibitors.

[0191] 9. A method of preventing or reducing a cancer in a subject in need thereof relative to a control, comprising administering to the subject (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonist comprising or consisting of bosentan or a derivative thereof; and (c) one or more immune checkpoint inhibitors.

[0192] 10. A method of reducing a number of cancer cells in a subject in need thereof relative to a control, comprising administering to the subject (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonist comprising or consisting of bosentan or a derivative thereof; and (c) one or more immune checkpoint inhibitors.

[0193] 11. A method of preventing or reducing a tumor volume in a subject in need thereof relative to a control, comprising administering to the subject (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonist comprising or consisting of bosentan or a derivative thereof; and (c) one or more immune checkpoint inhibitors.

[0194] 12. A method of preventing or reducing a cancer metastasis in a subject in need thereof relative to a control, comprising administering to the subject (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonist comprising or consisting of bosentan or a derivative thereof; and (c) one or more immune checkpoint inhibitors.

[0195] 13. The method of any one of embodiments 1-12, wherein the T cell composition comprises one or more of a CD3+T cell population, a 0D2+T cell population, or a OD28+T cell population.

[0196] 14. The method of any one of embodiments 1-13, wherein the T cell composition comprises or consists of a (i) a CD3+T cell population, (ii) a CD3+CD28+T cell population, (iii) a CD3+CD2+T cell population, or (iv) a CD3+CD2+CD28+T cell population.

[0197] 15. The method of embodiment 13 or 14, wherein the T cell population comprises one or more T cells.

[0198] 16. The method of any one of embodiments 1 -15, wherein the method reduces a number of cancer cells in the subject, relative to the control.

[0199] 17. The method of embodiment 16, wherein the number of cancer cells comprises an increase in a cancer cell size or an increase in a cancer cell number, relative to baseline.

[0200] 18. The method of any one of embodiments 1 -17, wherein the method prevents or reduces a tumor volume in the subject, relative to the control.

[0201] 19. The method of any one of embodiments 1 -18, wherein the method prevents or reduces a cancer metastasis in the subject, relative to the control.

[0202] 20. The method of any one of embodiments 1 -19, wherein the tumor or cancer comprises a solid tumor.

[0203] 21. The method of embodiment 20, wherein the cancer comprises a metastatic cancer.

[0204] 22. The method of embodiment 20 or 21 , wherein the cancer is a breast cancer.

[0205] 23. The method of embodiment 22, wherein the breast cancer is a HER2 positive breast cancer.

[0206] 24. The method of embodiment 22, wherein the breast cancer is a HER2 negative breast cancer.

[0207] 25. The method of embodiment 22, wherein the breast cancer is aHER2 / neu-negative breast cancer.

[0208] 26. The method of any one of embodiments 22, 24, or 25, wherein the breast cancer is a triple negative breast cancer.

[0209] 27. The method of any one of embodiments 22-26, wherein the breast cancer is selected from the group consisting of a stage I breast cancer, a stage II breast cancer, a stage III breast cancer, and a stage IV breast cancer.

[0210] 28. The method of any one of embodiments 1 -27, wherein preventing the cancer, the number of cancer cells, the tumor volume, or the cancer metastasis comprises preventing an advancement of a cancer stage or a cancer grade.

[0211] 29. The method of embodiment 28, wherein the advancement of the cancer stage comprises a stage 0 cancer progressing to a stage I cancer, a stage I cancer progressing to a stage II cancer, a stage II cancer progressing to a stage III cancer, or a stage III cancer progressing to a stage IV cancer.

[0212] 30. The method of embodiment 28, wherein the advancement of the cancer grade comprises a grade 1 cancer progressing to a grade 2 cancer, a grade 2 cancer progressing to a grade 3 cancer, or a grade 3 cancer progressing to a grade 4 cancer.

[0213] 31. The method of embodiment 16, wherein the number of cancer cells is reduced by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100%, relative to a control.

[0214] 32. The method of any one of embodiments 3, 7, 11 , 18, or 28, wherein the tumor volume is reduced by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100%, relative to a control.

[0215] 33. The method of any one of embodiments 4, 8, 12, 19, or 28, wherein the cancer metastasis is reduced by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100%, relative to a control.

[0216] 34. The method of any one of embodiments 4, 8, 12, 19, 28, or 33, wherein the reduction or prevention in metastasis comprises a reduction in one or more of a number of metastasis, a size of metastasis, a rate of metastasis growth, a recurrence of metastasis after a cancer therapy.

[0217] 35. The method of any one of embodiments 1-34, wherein the T cell composition is administered at a dose comprising at least about of 5 x 104T cells to at least about 15 x 1 O10cells.

[0218] 36. The method of any one of embodiments 1 -4, further comprising administering one or more immune checkpoint inhibitors to the subject.

[0219] 37. The method of any one of embodiments 5-25, wherein the one or more immune checkpoint inhibitors is selected from the group consisting of a CTLA-4 inhibitor, a PD-1 inhibitor, a PD-L1 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor, a TIGIT inhibitor, a VISTA inhibitor, a VEGFR1 inhibitor, a VEGFR2 inhibitor, and a VEGFR3 inhibitor.

[0220] 38. The method of any one of embodiments 5-37, wherein the one or more immune checkpoint inhibitors comprises an antibody.

[0221] 39. The method of embodiment 38, wherein the antibody is selected from the group consisting of pembrolizumab, ipilimumab, nivolumab, atezolizumab, cemiplimab, avelumab, durvalumab, and tremelimumab.

[0222] 40. The method of any one of embodiments 5-39, wherein the one or more immune checkpoint inhibitors are administered at a dose of about 0.5 mg / kg to about 10 mg / kg.

[0223] 41. The method of any one of embodiments 1 -8, wherein the endothelin receptor antagonist comprises bosentan or a derivative thereof.

[0224] 42. The method of any one of embodiments 9-41 , wherein the derivative of bosentan comprises a bosentan salt.

[0225] 43. The method of any one of embodiments 1 -42, wherein the endothelin receptor antagonist is administered at a dose of about 0.5 mg to about 600 mg.

[0226] 44. The method of any one of embodiments 5-43, wherein a first dose of the T cell composition is administered to the subject prior to administration of a first dose the one or more immune checkpoint inhibitors.

[0227] 45. The method of any one of embodiments 5-44, wherein a first dose of the one or more immune checkpoint inhibitors is administered to the subject prior to a first dose of the T cell composition.

[0228] 46. The method of any one of embodiments 5-44, wherein the subject is administered at least about three doses one or more immune checkpoint inhibitors.

[0229] 47. The method of embodiment 46, wherein the three or more doses of the one or more immune checkpoint inhibitors are administered to the subject about every three weeks to about every four weeks.

[0230] 48. The method of any one of embodiments 1 -47, wherein a first dose of the endothelin receptor antagonist is administered to the subject prior to administration of a first dose the T cell composition.

[0231] 49. The method of embodiment 48, wherein a second dose of the endothelin receptor antagonist is administered to the subject prior to administration of the first dose of the T cell composition.

[0232] 50. The method of any one of embodiments 1 -49, wherein the subject is administered the endothelin receptor antagonist daily.

[0233] 51 . The method of any one of embodiments 1 -50, wherein the one or more cancer antigens are selected from the group consisting of MAGE-A4, MAGEA1 , NY-ESO-1 , KK-LC-1 , FRAME, B7-H3, B7-H4, CD19, and CD22.

[0234] 52. The method of any one of embodiments 1-51 , wherein the T cell composition has an increase in one or more gene expression levels selected from the group consisting of a perforin-1 (PRF1 ) gene expression level, a granzyme B (GZMB) gene expression level, an interferon gamma (I N Fy) gene expression level, a tumor necrosis factor alpha (TNFa) gene expression level, a human leukocyte antigen class II histocompatibility antigen, DR alpha chain (HLA-DRA) gene expression level, a CD25 gene expression level, a CD69 gene expression level, and an IL-2 gene expression level, relative to a control.

[0235] 53. The method of any one of embodiments 1-52, wherein the T cell composition has an increase in one or more protein levels selected from the group consistingof a PRF1 protein level, a GZMB protein level, an INFy protein level, a TNFa protein level, an HLD-DRA protein level, a CD25 protein level, a CD69 protein level, and an IL-2 protein level, relative to a control.

[0236] 54. The method of any one of embodiments 1 -53, wherein at least about5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100% of the T cells in the T cell composition are specific to the one or more cancer antigens.

[0237] 55. The method of any one of embodiments 1-54, wherein the T cell composition comprises T cells that are derived from a subject having a cancer.

[0238] 56. The method of embodiment 55, wherein the T cells that are derived from a subject having cancer comprises T cells that were isolated or obtained from the subject having cancer.

[0239] 57. The method of embodiment 55 or 56, wherein the cancer comprises a solid tumor.

[0240] 58. The method of any one of embodiments 55-57, wherein the cancer comprises a metastatic cancer.

[0241] 59. The method of any one of embodiments 55-58, wherein the cancer is a breast cancer.

[0242] 60. The method of embodiment 59, wherein the breast cancer is a HER2 positive breast cancer.

[0243] 61. The method of embodiment 59, wherein the breast cancer is a HER2 negative breast cancer.

[0244] 62. The method of embodiment 59, wherein the breast cancer is aHER2 / neu-negative breast cancer.

[0245] 63. The method of any one of embodiments 59, 61 , or 62, wherein the breast cancer is a triple negative breast cancer.

[0246] 64. The method of any one of embodiments 59-63, wherein the breast cancer is selected from the group consisting of a stage I breast cancer, a stage II breast cancer, a stage III breast cancer, and a stage IV breast cancer.

[0247] 65. The method of any one of embodiments 1 -64, wherein one or more of the T cell compositions, the endothelin receptor antagonist, or the immune checkpoint inhibitor are formulated for subcutaneous, intravenous, intramuscular, intracranial, intraarterial, intraventricular, intranodal, intratumor, or intraperitoneal administration.

[0248] 66. The method of any one of embodiments 1-65, wherein the T cell compositions is formulated for subcutaneous, intravenous, intramuscular, intracranial, intraarterial, intraventricular, intranodal, intratumor, or intraperitoneal administration.

[0249] 67. The method of embodiment 65 or 66, wherein one or more of the T cell compositions, the endothelin receptor antagonist, or the immune checkpoint inhibitor are formulated in a vaccine.

[0250] 68. The method of embodiment 65 or 66, wherein one or more of the T cell compositions, the endothelin receptor antagonist, or the immune checkpoint inhibitor are formulated in a nanoparticle.

[0251] 69. The method of any one of embodiments 1 -64, wherein the endothelin receptor antagonist or the immune checkpoint inhibitor is formulated for oral administration or intravenous administration.

[0252] 70. The method of any one of embodiments 1 -69, wherein the control comprises the subject at baseline.

[0253] 71. The method of any one of embodiments 1 -69, wherein the control comprises a subject that is not administered one or more of (a) T cell composition specific to one or more cancer antigens; (b) an endothelin receptor antagonist; and (c) one or more immune checkpoint inhibitors.

[0254] 72. The method of embodiment 71 , wherein the control comprises a subject that is administered (a) alone.

[0255] 73. The method of embodiment 71 , wherein the control comprises a subject that is administered (b) alone.

[0256] 74. The method of embodiment 71 , wherein the control comprises a subject that is administered (c) alone.

[0257] 75. The method of embodiment 71 , wherein the control comprises a subject administered (a) and (b), but not (c).

[0258] 76. The method of embodiment 71 , wherein the control comprises a subject administered (b) and (c), but not (a).

[0259] 77. The method of embodiment 71 , wherein the control comprises a subject administered (a) and (c), but not (b).

[0260] 78. The method of any one of embodiments 71 -77, wherein the endothelin receptor antagonist comprises bosentan.

[0261] 79. Use of (a) a T cell composition specific to at least a portion of one or more cancer antigens; and (b) an endothelin receptor antagonist to prevent or reduce a cancer in a subject in need thereof relative to a control.

[0262] 80. Use of (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; and b) an endothelin receptor antagonist to reduce a number of cancer cells in a subject in need thereof relative to a control.

[0263] 81. Use of (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; and (b) an endothelin receptor antagonist to prevent or reduce a tumor volume in a subject in need thereof relative to a control.

[0264] 82. Use of (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; and (b) an endothelin receptor antagonist to prevent or reduce cancer metastasis in a subject in need thereof relative to a control.

[0265] 83. Use of (a) a T cell composition specific to one or more cancer antigens;(b) an endothelin receptor antagonist; and (c) one or more immune checkpoint inhibitors to prevent or reduce a cancer in a subject in need thereof relative to a control.

[0266] 84. Use of (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptorantagonist; and (c) one or more immune checkpoint inhibitors to prevent or reduce a number of cancer cells in a subject in need thereof relative to a control.

[0267] 85. Use of (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonist; and (c) one or more immune checkpoint inhibitors to prevent or reduce a tumor volume in a subject in need thereof relative to a control.

[0268] 86. Use of (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonist; and (c) one or more immune checkpoint inhibitors to prevent or reduce a cancer metastasis in a subject in need thereof relative to a control.

[0269] 87. Use of (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonist comprising or consisting of bosentan or a derivative thereof; and (c) one or more immune checkpoint inhibitors to prevent or reduce a cancer in a subject in need thereof relative to a control.

[0270] 88. Use of (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonist comprising or consisting of bosentan or a derivative thereof; and (c) one or more immune checkpoint inhibitors to reduce a number of cancer cells in a subject in need thereof relative to a control.

[0271] 89. Use of (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonist comprising or consisting of bosentan or a derivative thereof; and (c) one or more immune checkpoint inhibitors to prevent or reduce a tumor volume in a subject in need thereof relative to a control.

[0272] 90. Use of (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonist comprising or consisting of bosentan or a derivative thereof; and (c) one or moreimmune checkpoint inhibitors to prevent or reduce a cancer metastasis in a subject in need thereof relative to a control.

[0273] 91. The use of any one of embodiments 79-90, wherein the T cell composition comprises one or more of a CD3+T cell population, a CD2+T cell population, or a CD28+T cell population.

[0274] 92. The use of any one of embodiments 79-91 , wherein the T cell composition comprises or consists of a (i) a CD3+T cell population, (ii) a CD3+CD28+T cell population, (iii) a CD3+CD2+T cell population, or (iv) a CD3+CD2+CD28+T cell population.

[0275] 93. The use of embodiments 91 or 92, wherein the T cell population comprises one or more T cells.

[0276] 94. The use of any one of embodiments 79-93, wherein the subject experiences a reduction in a number of cancer cells relative to the control.

[0277] 95. The use of embodiment 94, wherein the number of cancer cells comprises an increase in a cancer cell size or an increase in a cancer cell number, relative to baseline.

[0278] 96. The use of any one of embodiments 79-95, wherein the subject experiences a reduced tumor volume relative to the control.

[0279] 97. The use of any one of embodiments 79-96, wherein the subject experiences a prevention or reduction in a cancer metastasis relative to the control.

[0280] 98. The use of any one of embodiments 79-97, wherein the tumor or cancer comprises a solid tumor.

[0281] 99. The use of embodiment 98, wherein the cancer comprises a metastatic cancer.

[0282] 100. The use of embodiments 98 or 99, wherein the cancer is a breast cancer.

[0283] 101. The use of embodiment 100, wherein the breast cancer is a HER2 positive breast cancer.

[0284] 102. The use of embodiment 100, wherein the breast cancer is a HER2 negative breast cancer.

[0285] 103. The use of embodiment 100, wherein the breast cancer is a HER2 / neu- negative breast cancer.

[0286] 104. The use of any one of embodiments 100, 102, or 103, wherein the breast cancer is a triple negative breast cancer.

[0287] 105. The use of any one of embodiments 100-104, wherein the breast cancer is selected from the group consisting of a stage I breast cancer, a stage II breast cancer, a stage III breast cancer, and a stage IV breast cancer.

[0288] 106. The use of any one of embodiments 79-105, wherein preventing the cancer, the number of cancer cells, the tumor volume, or the cancer metastasis comprises preventing an advancement of a cancer stage or a cancer grade.

[0289] 107. The use of embodiment 106, wherein the advancement of the cancer stage comprises a stage 0 cancer progressing to a stage I cancer, a stage I cancer progressing to a stage II cancer, a stage II cancer progressing to a stage III cancer, or a stage III cancer progressing to a stage IV cancer.

[0290] 108. The use of embodiment 106, wherein the advancement of the cancer grade comprises a grade 1 cancer progressing to a grade 2 cancer, a grade 2 cancer progressing to a grade 3 cancer, or a grade 3 cancer progressing to a grade 4 cancer.

[0291] 109. The use of embodiment 94, wherein the number of cancer cells is reduced by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100%, relative to a control.

[0292] 110. The use of any one of embodiments 81 , 85, 89, and 96, wherein the tumor volume is reduced by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100%, relative to a control.

[0293] 11 1. The use of any one of embodiments 82, 86, 90, and 97, wherein the cancer metastasis is reduced by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100%, relative to a control.

[0294] 112. The use of any one of embodiments 82, 86, 90, 97, and 1 1 1 , wherein the reduction or prevention in metastasis comprises a reduction in one or more of a number of metastasis, a size of metastasis, a rate of metastasis growth, a recurrence of metastasis after a cancer therapy.

[0295] 113. The use of any one of embodiments 79-112, wherein the T cell composition is administered at a dose comprising at least about of 5 x 104T cells to at least about 15 x 1 O10cells.

[0296] 114. The use of any one of embodiments 79-82, wherein one or more immune checkpoint inhibitors are administered to the subject.

[0297] 115. The use of any one of embodiments 83-1 14, wherein the one or more immune checkpoint inhibitors is selected from the group consisting of a CTLA-4 inhibitor, a PD-1 inhibitor, a PD-L1 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor, a TIGIT inhibitor, a VISTA inhibitor, a VEGFR1 inhibitor, a VEGFR2 inhibitor, and a VEGFR3 inhibitor.

[0298] 116. The use of any one of embodiments 83-1 15, wherein the one or more immune checkpoint inhibitors comprises an antibody.

[0299] 117. The use of embodiment 1 16, wherein the antibody is selected from the group consisting of pembrolizumab, ipilimumab, nivolumab, atezolizumab, cemiplimab, avelumab, durvalumab, and tremelimumab.

[0300] 118. The use of any one of embodiments 83-1 17, wherein the one or more immune checkpoint inhibitors are administered at a dose of about 0.5 mg / kg to about 10 mg / kg.

[0301] 119. The use of any one of embodiments 79-86, wherein the endothelin receptor antagonist comprises bosentan or a derivative thereof.

[0302] 120. The use of any one of embodiments 87-1 19, wherein the derivative of bosentan comprises a bosentan salt.

[0303] 121. The use of any one of embodiments 79-120, wherein the endothelin receptor antagonist is administered at a dose of about 0.5 mg to about 600 mg.

[0304] 122. The use of any one of embodiments 83-121 , wherein a first dose of theT cell composition is administered to the subject prior to administration of a first dose the one or more immune checkpoint inhibitors.

[0305] 123. The use of any one of embodiments 83-121 , wherein a first dose of the one or more immune checkpoint inhibitors is administered to the subject prior to a first dose of the T cell composition.

[0306] 124. The use of any one of embodiments 83-121 , wherein the subject is administered at least about three doses one or more immune checkpoint inhibitors.

[0307] 125. The use of embodiment 124, wherein the three or more doses of the one or more immune checkpoint inhibitors are administered to the subject about every three weeks to about every four weeks.

[0308] 126. The use of any one of embodiments 79-125, wherein a first dose of the endothelin receptor antagonist is administered to the subject prior to administration of a first dose the T cell composition.

[0309] 127. The use of embodiment 126, wherein a second dose of the endothelin receptor antagonist is administered to the subject prior to administration of the first dose of the T cell composition.

[0310] 128. The use of any one of embodiments 79-127, wherein the subject is administered the endothelin receptor antagonist daily.

[0311] 129. The use of any one of embodiments 79-128, wherein the one or more cancer antigens are selected from the group consisting of MAGE-A4, MAGEA1 , NY-ESO-1 , KK-LC-1 , FRAME, B7-H3, B7-H4, CD19, and CD22.

[0312] 130. The use of any one of embodiments 79-129, wherein the T cell composition has an increase in one or more gene expression levels selected from the group consisting of a perforin-1 (PRF1 ) gene expression level, a granzyme B (GZMB) gene expression level, an interferon gamma (INFy) gene expression level, a tumor necrosis factor alpha (TNFa) gene expression level, a human leukocyte antigen class II histocompatibilityantigen, DR alpha chain (HLA-DRA) gene expression level, a OD25 gene expression level, a CD69 gene expression level, and an IL-2 gene expression level, relative to a control.

[0313] 131. The use of any one of embodiments 79-130, wherein the T cell composition has an increase in one or more protein levels selected from the group consisting of a PRF1 protein level, a GZMB protein level, an INFy protein level, a TNFa protein level, an HLD-DRA protein level, a CD25 protein level, a CD69 protein level, and an IL-2 protein level, relative to a control.

[0314] 132. The use of any one of embodiments 79-131 , wherein at least about 5%,10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100% of the T cells in the T cell composition are specific to the one or more cancer antigens.

[0315] 133. The use of any one of embodiments 79-132, wherein the T cell composition comprises T cells that are derived from a subject having a cancer.

[0316] 134. The use of any one of embodiments 79-132, wherein the T cell composition comprises T cells that are derived from a subject having a cancer.

[0317] 135. The use of embodiment 133 or 134, wherein the cancer comprises a solid tumor.

[0318] 136. The use of any one of embodiments 133-135, wherein the cancer comprises a metastatic cancer.

[0319] 137. The use of any one of embodiments 133-136, wherein the cancer is a breast cancer.

[0320] 138. The use of embodiment 137, wherein the breast cancer is a HER2 positive breast cancer.

[0321] 139. The use of embodiment 137, wherein the breast cancer is a HER2 negative breast cancer.

[0322] 140. The use of embodiment 137, wherein the breast cancer is a HER2 / neu- negative breast cancer.

[0323] 141 . The use of any one of embodiments 137, 139, or 140 wherein the breast cancer is a triple negative breast cancer.

[0324] 142. The use of any one of embodiments 137-141 , wherein the breast cancer is selected from the group consisting of a stage I breast cancer, a stage II breast cancer, a stage III breast cancer, and a stage IV breast cancer.

[0325] 143. The use of any one of embodiments 79-142, wherein one or more of the T cell compositions, the endothelin receptor antagonist, or the immune checkpoint inhibitor are formulated for subcutaneous, intravenous, intramuscular, intracranial, intraarterial, intraventricular, intranodal, intratumor, or intraperitoneal administration.

[0326] 144. The use of any one of embodiments 79-143, wherein the T cell compositions is formulated for subcutaneous, intravenous, intramuscular, intracranial, intraarterial, intraventricular, intranodal, intratumor, or intraperitoneal administration.

[0327] 145. The use of embodiment 143 or 144, wherein one or more of the T cell compositions, the endothelin receptor antagonist, or the immune checkpoint inhibitor are formulated in a vaccine.

[0328] 146. The use of embodiment 143 or 144, wherein one or more of the T cell compositions, the endothelin receptor antagonist, or the immune checkpoint inhibitor are formulated in a nanoparticle.

[0329] 147. The use of any one of embodiments 79-146, wherein the endothelin receptor antagonist or the immune checkpoint inhibitor is formulated for oral administration or intravenous administration.

[0330] 148. The use of any one of embodiments 79-147, wherein the control comprises the subject at baseline.

[0331] 149. The use of any one of embodiments 79-148, wherein the control comprises a subject that is not administered one or more of (a) T cell composition specific to one or more cancer antigens; (b) an endothelin receptor antagonist; and(c) one or more immune checkpoint inhibitors.

[0332] 150. The use of embodiment 149, wherein the control comprises a subject that is administered (a) alone.

[0333] 151. The use of embodiment 149, wherein the control comprises a subject that is administered (b) alone.

[0334] 152. The use of embodiment 149, wherein the control comprises a subject that is administered (c) alone.

[0335] 153. The use of embodiment 149, wherein the control comprises a subject administered (a) and (b), but not (c).

[0336] 154. The use of embodiment 149, wherein the control comprises a subject administered (b) and (c), but not (a).

[0337] 155. The use of embodiment 149, wherein the control comprises a subject administered (a) and (c), but not (b).

[0338] 156. The use of any one of embodiments 79-155, wherein the endothelin receptor antagonist comprises bosentan.

Claims

1. CLAIMS l / We claim:

1. A method of preventing or reducing a cancer in a subject in need thereof relative to a control, comprising administering to the subject(a) a T cell composition specific to at least a portion of one or more cancer antigens; and(b) an endothelin receptor antagonist.

2. A method of reducing a number of cancer cells in a subject in need thereof relative to a control, comprising administering to the subject(a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; and(b) an endothelin receptor antagonist.

3. A method of preventing or reducing a tumor volume in a subject in need thereof relative to a control, comprising administering to the subject(a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; and(b) an endothelin receptor antagonist.

4. A method of preventing or reducing cancer metastasis in a subject in need thereof relative to a control, comprising administering to the subject(a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; and(b) an endothelin receptor antagonist.

5. A method of preventing or reducing a cancer in a subject in need thereof relative to a control, comprising administering to the subject(a) a T cell composition specific to one or more cancer antigens;(b) an endothelin receptor antagonist; and(c) one or more immune checkpoint inhibitors.

6. A method of preventing or reducing a number of cancer cells in a subject in need thereof relative to a control, comprising administering to the subject(a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens;(b) an endothelin receptor antagonist; and(c) one or more immune checkpoint inhibitors.

7. A method of preventing or reducing a tumor volume in a subject in need thereof relative to a control, comprising administering to the subject(a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens;(b) an endothelin receptor antagonist; and(c) one or more immune checkpoint inhibitors.

8. A method of preventing or reducing a cancer metastasis in a subject in need thereof relative to a control, comprising administering to the subject(a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens;(b) an endothelin receptor antagonist; and(c) one or more immune checkpoint inhibitors.

9. A method of preventing or reducing a cancer in a subject in need thereof relative to a control, comprising administering to the subject(a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens;(b) an endothelin receptor antagonist comprising or consisting of bosentan or a derivative thereof; and(c) one or more immune checkpoint inhibitors.

10. A method of reducing a number of cancer cells in a subject in need thereof relative to a control, comprising administering to the subject(a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens;(b) an endothelin receptor antagonist comprising or consisting of bosentan or a derivative thereof; and(c) one or more immune checkpoint inhibitors.1 1. A method of preventing or reducing a tumor volume in a subject in need thereof relative to a control, comprising administering to the subject(a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens;(b) an endothelin receptor antagonist comprising or consisting of bosentan or a derivative thereof; and(c) one or more immune checkpoint inhibitors.

12. A method of preventing or reducing a cancer metastasis in a subject in need thereof relative to a control, comprising administering to the subject(a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens;(b) an endothelin receptor antagonist comprising or consisting of bosentan or a derivative thereof; and(c) one or more immune checkpoint inhibitors.

13. The method of any one of claims 1 -12, wherein the T cell composition comprises one or more of a CD3+T cell population, a CD2+T cell population, or a OD28+T cell population.

14. The method of any one of claims 1 -13, wherein the T cell composition comprises or consists of a (i) a CD3+T cell population, (ii) a 0D3+CD28+T cell population, (iii) a CD3+CD2+T cell population, or (iv) a CD3+CD2+CD28+T cell population.

15. The method of claim 13 or 14, wherein the T cell population comprises one or more T cells.

16. The method of any one of claims 1 -15, wherein the method reduces a number of cancer cells in the subject, relative to the control.

17. The method of claim 16, wherein the number of cancer cells comprises an increase in a cancer cell size or an increase in a cancer cell number, relative to baseline.

18. The method of any one of claims 1 -17, wherein the method prevents or reduces a tumor volume in the subject, relative to the control.

19. The method of any one of claims 1 -18, wherein the method prevents or reduces a cancer metastasis in the subject, relative to the control.

20. The method of any one of claims 1 -19, wherein the tumor or cancer comprises a solid tumor.21 . The method of claim 20, wherein the cancer comprises a metastatic cancer.

22. The method of claim 20 or 21 , wherein the cancer is a breast cancer.

23. The method of claim 22, wherein the breast cancer is a HER2 positive breast cancer.

24. The method of claim 22, wherein the breast cancer is a HER2 negative breast cancer.

25. The method of claim 22, wherein the breast cancer is a HER2 / neu-negative breast cancer.

26. The method of any one of claims 22, 24, or 25, wherein the breast cancer is a triple negative breast cancer.

27. The method of any one of claims 22-26, wherein the breast cancer is selected from the group consisting of a stage I breast cancer, a stage II breast cancer, a stage III breast cancer, and a stage IV breast cancer.

28. The method of any one of claims 1 -27, wherein preventing the cancer, the number of cancer cells, the tumor volume, or the cancer metastasis comprises preventing an advancement of a cancer stage or a cancer grade.

29. The method of claim 28, wherein the advancement of the cancer stage comprises a stage 0 cancer progressing to a stage I cancer, a stage I cancer progressing to a stage II cancer, a stage II cancer progressing to a stage III cancer, or a stage III cancer progressing to a stage IV cancer.

30. The method of claim 28, wherein the advancement of the cancer grade comprises a grade 1 cancer progressing to a grade 2 cancer, a grade 2 cancer progressing to a grade 3 cancer, or a grade 3 cancer progressing to a grade 4 cancer.31 . The method of claim 16, wherein the number of cancer cells is reduced by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100%, relative to a control.

32. The method of any one of claims 3, 7, 1 1 , 18, or 28, wherein the tumor volume is reduced by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100%, relative to a control.

33. The method of any one of claims 4, 8, 12, 19, or 28, wherein the cancer metastasis is reduced by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100%, relative to a control.

34. The method of any one of claims 4, 8, 12, 19, 28, or 33, wherein the reduction or prevention in metastasis comprises a reduction in one or more of a number of metastasis, a size of metastasis, a rate of metastasis growth, a recurrence of metastasis after a cancer therapy.

35. The method of any one of claims 1 -34, wherein the T cell composition is administered at a dose comprising at least about of 5 x 104T cells to at least about 15 x 1010cells.

36. The method of any one of claims 1 -4, further comprising administering one or more immune checkpoint inhibitors to the subject.

37. The method of any one of claims 5-25, wherein the one or more immune checkpoint inhibitors is selected from the group consisting of a CTLA-4 inhibitor, a PD-1 inhibitor, a PD-L1 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor, a TIGIT inhibitor, a VISTA inhibitor, a VEGFR1 inhibitor, a VEGFR2 inhibitor, and a VEGFR3 inhibitor.

38. The method of any one of claims 5-37, wherein the one or more immune checkpoint inhibitors comprises an antibody.

39. The method of claim 38, wherein the antibody is selected from the group consisting of pembrolizumab, ipilimumab, nivolumab, atezolizumab, cemiplimab, avelumab, durvalumab, and tremelimumab.

40. The method of any one of claims 5-39, wherein the one or more immune checkpoint inhibitors are administered at a dose of about 0.5 mg / kg to about 10 mg / kg.

41. The method of any one of claims 1 -8, wherein the endothelin receptor antagonist comprises bosentan or a derivative thereof.

42. The method of any one of claims 9-41 , wherein the derivative of bosentan comprises a bosentan salt.

43. The method of any one of claims 1 -42, wherein the endothelin receptor antagonist is administered at a dose of about 0.5 mg to about 600 mg.

44. The method of any one of claims 5-43, wherein a first dose of the T cell composition is administered to the subject prior to administration of a first dose the one or more immune checkpoint inhibitors.

45. The method of any one of claims 5-44, wherein a first dose of the one or more immune checkpoint inhibitors is administered to the subject prior to a first dose of the T cell composition.

46. The method of any one of claims 5-44, wherein the subject is administered at least about three doses one or more immune checkpoint inhibitors.

47. The method of claim 46, wherein the three or more doses of the one or more immune checkpoint inhibitors are administered to the subject about every three weeks to about every four weeks.

48. The method of any one of claims 1 -47, wherein a first dose of the endothelin receptor antagonist is administered to the subject prior to administration of a first dose the T cell composition.

49. The method of claim 48, wherein a second dose of the endothelin receptor antagonist is administered to the subject prior to administration of the first dose of the T cell composition.

50. The method of any one of claims 1 -49, wherein the subject is administered the endothelin receptor antagonist daily.

51. The method of any one of claims 1 -50, wherein the one or more cancer antigens are selected from the group consisting of MAGE-A4, MAGEA1 , NY-ESO-1 , KK- LC-1 , FRAME, B7-H3, B7-H4, CD19, and OD22.

52. The method of any one of claims 1 -51 , wherein the T cell composition has an increase in one or more gene expression levels selected from the group consisting of a perforin-1 (PRF1 ) gene expression level, a granzyme B (GZMB) gene expression level, an interferon gamma (I N Fy) gene expression level, a tumor necrosis factor alpha (TNFa) gene expression level, a human leukocyte antigen class II histocompatibility antigen, DR alpha chain (HLA-DRA) gene expression level, a CD25 gene expression level, a CD69 gene expression level, and an IL-2 gene expression level, relative to a control.

53. The method of any one of claims 1 -52, wherein the T cell composition has an increase in one or more protein levels selected from the group consisting of a PRF1 protein level, a GZMB protein level, an INFy protein level, a TNFa protein level, an HLD-DRA protein level, a CD25 protein level, a CD69 protein level, and an IL-2 protein level, relative to a control.

54. The method of any one of claims 1 -53, wherein at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100% of the T cells in the T cell composition are specific to the one or more cancer antigens.

55. The method of any one of claims 1 -54, wherein the T cell composition comprises T cells that are derived from a subject having a cancer.

56. The method of claim 55, wherein the T cells that are derived from a subject having cancer comprises T cells that were isolated or obtained from the subject having cancer.

57. The method of claim 55 or 56, wherein the cancer comprises a solid tumor.

58. The method of any one of claims 55-57, wherein the cancer comprises a metastatic cancer.

59. The method of any one of claims 55-58, wherein the cancer is a breast cancer.

60. The method of claim 59, wherein the breast cancer is a HER2 positive breast cancer.61 . The method of claim 59, wherein the breast cancer is a HER2 negative breast cancer.

62. The method of claim 59, wherein the breast cancer is a HER2 / neu-negative breast cancer.

63. The method of any one of claims 59, 61 , or 62, wherein the breast cancer is a triple negative breast cancer.

64. The method of any one of claims 59-63, wherein the breast cancer is selected from the group consisting of a stage I breast cancer, a stage II breast cancer, a stage III breast cancer, and a stage IV breast cancer.

65. The method of any one of claims 1 -64, wherein one or more of the T cell compositions, the endothelin receptor antagonist, or the immune checkpoint inhibitor areformulated for subcutaneous, intravenous, intramuscular, intracranial, intra-arterial, intraventricular, intranodal, intratumor, or intraperitoneal administration.

66. The method of any one of claims 1 -65, wherein the T cell compositions is formulated for subcutaneous, intravenous, intramuscular, intracranial, intra-arterial, intraventricular, intranodal, intratumor, or intraperitoneal administration.

67. The method of claim 65 or 66, wherein one or more of the T cell compositions, the endothelin receptor antagonist, or the immune checkpoint inhibitor are formulated in a vaccine.

68. The method of claim 65 or 66, wherein one or more of the T cell compositions, the endothelin receptor antagonist, or the immune checkpoint inhibitor are formulated in a nanoparticle.

69. The method of any one of claims 1 -64, wherein the endothelin receptor antagonist or the immune checkpoint inhibitor is formulated for oral administration or intravenous administration.

70. The method of any one of claims 1 -69, wherein the control comprises the subject at baseline.71 . The method of any one of claims 1 -69, wherein the control comprises a subject that is not administered one or more of(a) T cell composition specific to one or more cancer antigens;(b) an endothelin receptor antagonist; and(c) one or more immune checkpoint inhibitors.

72. The method of claim 71 , wherein the control comprises a subject that is administered (a) alone.

73. The method of claim 71 , wherein the control comprises a subject that is administered (b) alone.

74. The method of claim 71 , wherein the control comprises a subject that is administered (c) alone.

75. The method of claim 71 , wherein the control comprises a subject administered(a) and (b), but not (c).

76. The method of claim 71 , wherein the control comprises a subject administered(b) and (c), but not (a). in. The method of claim 71 , wherein the control comprises a subject administered (a) and (c), but not (b).

78. The method of any one of claims 71 -77, wherein the endothelin receptor antagonist comprises bosentan.

79. Use of (a) a T cell composition specific to at least a portion of one or more cancer antigens; and (b) an endothelin receptor antagonist to prevent or reduce a cancer in a subject in need thereof relative to a control.

80. Use of (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; and b) an endothelin receptor antagonist to reduce a number of cancer cells in a subject in need thereof relative to a control.81 . Use of (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; and (b) an endothelin receptor antagonist to prevent or reduce a tumor volume in a subject in need thereof relative to a control.

82. Use of (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; and (b) an endothelin receptor antagonist to prevent or reduce cancer metastasis in a subject in need thereof relative to a control.

83. Use of (a) a T cell composition specific to one or more cancer antigens; (b) an endothelin receptor antagonist; and (c) one or more immune checkpoint inhibitors to prevent or reduce a cancer in a subject in need thereof relative to a control.

84. Use of (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonist; and (c) one or more immune checkpoint inhibitors to prevent or reduce a number of cancer cells in a subject in need thereof relative to a control.

85. Use of (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonist; and (c) one or more immune checkpoint inhibitors to prevent or reduce a tumor volume in a subject in need thereof relative to a control.

86. Use of (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonist; and (c) one or more immune checkpoint inhibitors to prevent or reduce a cancer metastasis in a subject in need thereof relative to a control.

87. Use of (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonist comprising or consisting of bosentan or a derivative thereof; and (c) one or more immune checkpoint inhibitors to prevent or reduce a cancer in a subject in need thereof relative to a control.

88. Use of (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonistcomprising or consisting of bosentan or a derivative thereof; and (c) one or more immune checkpoint inhibitors to reduce a number of cancer cells in a subject in need thereof relative to a control.

89. Use of (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonist comprising or consisting of bosentan or a derivative thereof; and (c) one or more immune checkpoint inhibitors to prevent or reduce a tumor volume in a subject in need thereof relative to a control.

90. Use of (a) a T cell composition comprising one or more receptors specific to at least a portion of one or more cancer antigens; (b) an endothelin receptor antagonist comprising or consisting of bosentan or a derivative thereof; and (c) one or more immune checkpoint inhibitors to prevent or reduce a cancer metastasis in a subject in need thereof relative to a control.91 . The use of any one of claims 79-90, wherein the T cell composition comprises one or more of a CD3+T cell population, a CD2+T cell population, or a CD28+T cell population.

92. The use of any one of claims 79-91 , wherein the T cell composition comprises or consists of a (i) a CD3+T cell population, (ii) a CD3+CD28+T cell population, (iii) a CD3+CD2+T cell population, or (iv) a CD3+CD2+CD28+T cell population.

93. The use of claims 91 or 92, wherein the T cell population comprises one or more T cells.

94. The use of any one of claims 79-93, wherein the subject experiences a reduction in a number of cancer cells relative to the control.

95. The use of claim 94, wherein the number of cancer cells comprises an increase in a cancer cell size or an increase in a cancer cell number, relative to baseline.

96. The use of any one of claims 79-95, wherein the subject experiences a reduced tumor volume relative to the control.

97. The use of any one of claims 79-96, wherein the subject experiences a prevention or reduction in a cancer metastasis relative to the control.

98. The use of any one of claims 79-97, wherein the tumor or cancer comprises a solid tumor.

99. The use of claim 98, wherein the cancer comprises a metastatic cancer.

100. The use of claims 98 or 99, wherein the cancer is a breast cancer.

101. The use of claim 100, wherein the breast cancer is a HER2 positive breast cancer.

102. The use of claim 100, wherein the breast cancer is a HER2 negative breast cancer.

103. The use of claim 100, wherein the breast cancer is a HER2 / neu-negative breast cancer.

104. The use of any one of claims 100, 102, or 103, wherein the breast cancer is a triple negative breast cancer.

105. The use of any one of claims 100-104, wherein the breast cancer is selected from the group consisting of a stage I breast cancer, a stage II breast cancer, a stage III breast cancer, and a stage IV breast cancer.

106. The use of any one of claims 79-105, wherein preventing the cancer, the number of cancer cells, the tumor volume, or the cancer metastasis comprises preventing an advancement of a cancer stage or a cancer grade.

107. The use of claim 106, wherein the advancement of the cancer stage comprises a stage 0 cancer progressing to a stage I cancer, a stage I cancer progressing to a stage II cancer, a stage II cancer progressing to a stage III cancer, or a stage III cancer progressing to a stage IV cancer.

108. The use of claim 106, wherein the advancement of the cancer grade comprises a grade 1 cancer progressing to a grade 2 cancer, a grade 2 cancer progressing to a grade 3 cancer, or a grade 3 cancer progressing to a grade 4 cancer.

109. The use of claim 94, wherein the number of cancer cells is reduced by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100%, relative to a control.1 10. The use of any one of claims 81 , 85, 89, and 96, wherein the tumor volume is reduced by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100%, relative to a control.1 1 1. The use of any one of claims 82, 86, 90, and 97, wherein the cancer metastasis is reduced by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100%, relative to a control.1 12. The use of any one of claims 82, 86, 90, 97, and 111 , wherein the reduction or prevention in metastasis comprises a reduction in one or more of a number of metastasis,a size of metastasis, a rate of metastasis growth, a recurrence of metastasis after a cancer therapy.1 13. The use of any one of claims 79-112, wherein the T cell composition is administered at a dose comprising at least about of 5 x 104T cells to at least about 15 x 1 O10cells.1 14. The use of any one of claims 79-82, wherein one or more immune checkpoint inhibitors are administered to the subject.1 15. The use of any one of claims 83-114, wherein the one or more immune checkpoint inhibitors is selected from the group consisting of a CTLA-4 inhibitor, a PD-1 inhibitor, a PD-L1 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor, a TIGIT inhibitor, a VISTA inhibitor, a VEGFR1 inhibitor, a VEGFR2 inhibitor, and a VEGFR3 inhibitor.1 16. The use of any one of claims 83-115, wherein the one or more immune checkpoint inhibitors comprises an antibody.1 17. The use of claim 116, wherein the antibody is selected from the group consisting of pembrolizumab, ipilimumab, nivolumab, atezolizumab, cemiplimab, avelumab, durvalumab, and tremelimumab.1 18. The use of any one of claims 83-117, wherein the one or more immune checkpoint inhibitors are administered at a dose of about 0.5 mg / kg to about 10 mg / kg.1 19. The use of any one of claims 79-86, wherein the endothelin receptor antagonist comprises bosentan or a derivative thereof.

120. The use of any one of claims 87-119, wherein the derivative of bosentan comprises a bosentan salt.

121. The use of any one of claims 79-120, wherein the endothelin receptor antagonist is administered at a dose of about 0.5 mg to about 600 mg.

122. The use of any one of claims 83-121 , wherein a first dose of the T cell composition is administered to the subject prior to administration of a first dose the one or more immune checkpoint inhibitors.

123. The use of any one of claims 83-121 , wherein a first dose of the one or more immune checkpoint inhibitors is administered to the subject prior to a first dose of the T cell composition.

124. The use of any one of claims 83-121 , wherein the subject is administered at least about three doses one or more immune checkpoint inhibitors.

125. The use of claim 124, wherein the three or more doses of the one or more immune checkpoint inhibitors are administered to the subject about every three weeks to about every four weeks.

126. The use of any one of claims 79-125, wherein a first dose of the endothelin receptor antagonist is administered to the subject prior to administration of a first dose the T cell composition.

127. The use of claim 126, wherein a second dose of the endothelin receptor antagonist is administered to the subject prior to administration of the first dose of the T cell composition.

128. The use of any one of claims 79-127, wherein the subject is administered the endothelin receptor antagonist daily.

129. The use of any one of claims 79-128, wherein the one or more cancer antigens are selected from the group consisting of MAGE-A4, MAGEA1 , NY-ESO-1 , KK-LC-1 , FRAME, B7-H3, B7-H4, CD19, and CD22.

130. The use of any one of claims 79-129, wherein the T cell composition has an increase in one or more gene expression levels selected from the group consisting of a perforin-1 (PRF1 ) gene expression level, a granzyme B (GZMB) gene expression level, an interferon gamma (INFy) gene expression level, a tumor necrosis factor alpha (TNFa) gene expression level, a human leukocyte antigen class II histocompatibility antigen, DR alpha chain (HLA-DRA) gene expression level, a OD25 gene expression level, a CD69 gene expression level, and an IL-2 gene expression level, relative to a control.

131. The use of any one of claims 79-130, wherein the T cell composition has an increase in one or more protein levels selected from the group consisting of a PRF1 protein level, a GZMB protein level, an INFy protein level, a TNFa protein level, an HLD-DRA protein level, a CD25 protein level, a CD69 protein level, and an IL-2 protein level, relative to a control.

132. The use of any one of claims 79-131 , wherein at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100% of the T cells in the T cell composition are specific to the one or more cancer antigens.

133. The use of any one of claims 79-132, wherein the T cell composition comprises T cells that are derived from a subject having a cancer.

134. The use of claim 133, wherein the T cells that are derived from a subject having cancer comprises T cells that were isolated or obtained from the subject having cancer.

135. The use of claim 133 or 134, wherein the cancer comprises a solid tumor.

136. The use of any one of claims 133-135, wherein the cancer comprises a metastatic cancer.

137. The use of any one of claims 133-136, wherein the cancer is a breast cancer.

138. The use of claim 137, wherein the breast cancer is a HER2 positive breast cancer.

139. The use of claim 137, wherein the breast cancer is a HER2 negative breast cancer.

140. The use of claim 137, wherein the breast cancer is a HER2 / neu-negative breast cancer.141 . The use of any one of claims 137, 139, or 140 wherein the breast cancer is a triple negative breast cancer.

142. The use of any one of claims 137-141 , wherein the breast cancer is selected from the group consisting of a stage I breast cancer, a stage II breast cancer, a stage III breast cancer, and a stage IV breast cancer.

143. The use of any one of claims 79-142, wherein one or more of the T cell compositions, the endothelin receptor antagonist, or the immune checkpoint inhibitor are formulated for subcutaneous, intravenous, intramuscular, intracranial, intra-arterial, intraventricular, intranodal, intratumor, or intraperitoneal administration.

144. The use of any one of claims 79-143, wherein the T cell compositions is formulated for subcutaneous, intravenous, intramuscular, intracranial, intra-arterial, intraventricular, intranodal, intratumor, or intraperitoneal administration.

145. The use of claim 143 or 144, wherein one or more of the T cell compositions, the endothelin receptor antagonist, or the immune checkpoint inhibitor are formulated in a vaccine.

146. The use of claim 143 or 144, wherein one or more of the T cell compositions, the endothelin receptor antagonist, or the immune checkpoint inhibitor are formulated in a nanoparticle.

147. The use of any one of claims 79-146, wherein the endothelin receptor antagonist or the immune checkpoint inhibitor is formulated for oral administration or intravenous administration.

148. The use of any one of claims 79-147, wherein the control comprises the subject at baseline.

149. The use of any one of claims 79-148, wherein the control comprises a subject that is not administered one or more of (a) T cell composition specific to one or more cancer antigens; (b) an endothelin receptor antagonist; and(c) one or more immune checkpoint inhibitors.

150. The use of claim 149, wherein the control comprises a subject that is administered (a) alone.

151. The use of claim 149, wherein the control comprises a subject that is administered (b) alone.

152. The use of claim 149, wherein the control comprises a subject that is administered (c) alone.

153. The use of claim 149, wherein the control comprises a subject administered(a) and (b), but not (c).

154. The use of claim 149, wherein the control comprises a subject administered(b) and (c), but not (a).

155. The use of claim 149, wherein the control comprises a subject administered (a) and (c), but not (b).

156. The use of any one of claims 79-155, wherein the endothelin receptor antagonist comprises bosentan.

Images