Methods and materials for treating cancer

Abstract

This document provides methods and materials involved in treating a mammal (e.g., a human) having cancer (e.g., a cancer including one or more solid tumors). For example, methods and materials provided herein can be used to sensitize a cancer to immune checkpoint blockade (ICB) (e.g., administration of one or more immune checkpoint inhibitors). For example, a mammal having cancer can be administered (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide to sensitize the cancer to ICB and, optionally, can also be administered one or more immune checkpoint inhibitors to treat the cancer. In another example, a mammal having cancer can be administered (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 to enhance cellular immunotherapy and, optionally, can also be administered one or more cellular immunotherapies to treat the cancer.

Description

[0001] METHODS AND MATERIALS FOR TREATING CANCER

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims the benefit of U.S. Patent Application Serial No. 63 / 561,050, filed on March 4, 2024. The disclosure of the prior application is considered part of (and is incorporated by reference in) the disclosure of this application.

[0004] TECHNICAL FIELD

[0005] This document relates to methods and materials involved in treating a mammal (e.g., a human) having cancer (e.g., a cancer including one or more solid tumors). For example, methods and materials provided herein can be used to sensitize a cancer to immune checkpoint blockade (ICB) (e.g., administration of one or more immune checkpoint inhibitors) and / or to enhance cellular immunotherapy (e.g., enhance effectiveness of administration of immune cells such as tumor-infiltrating lymphocytes (TILs)). For example, a mammal (e.g., a human) having cancer can be administered (a) one or more inhibitors of a nitric oxide synthase (NOS) 2 polypeptide and (b) one or more inhibitors of a cyclooxygenase (COX) 2 polypeptide to sensitize the cancer to ICB and, optionally, can also be administered one or more immune checkpoint inhibitors to treat the cancer. In another example, a mammal (e.g., a human) having cancer can be administered (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 to enhance cellular immunotherapy and, optionally, can also be administered one or more cellular immunotherapies (e.g., engineered or expanded TILs) to treat the cancer.

[0006] BACKGROUND

[0007] Hepatocellular carcinoma (HCC) is the fastest rising cause of cancer-related death in the US and is the second leading cause of cancer death in East Asia and sub-Saharan Africa and the sixth most common in western countries. The incidence of HCC will continue rise as hepatitis C and non-alcoholic steatohepatitis (NASH) as well as obesity become more prevalent in the United States. HCC is complex, and treatment strategies need to be tailored to the underlying liver disease which often involves sustained inflammatory changes. For early-stage HCC, the treatment of choice remains resection; however, the majority of patients (up to 70%) are found to have unresectable disease upon initial presentation. See, e.g., Leowattana et al., World J. Gastroenterol., 29( 10): 1551 -1568 (2023).

[0008] The IMBRAVE150 study, a phase 3, open label, multicenter randomized trial, compared atezolizumab combined with bevacizumab vs sorafenib in patients with unresectable HCC. The dual combination resulted in better overall and progression-free survival. However, the progression-free survival only increased from 4.3 to 6.8 months and the 12-month overall survival only increased from 56% to 67% (Finn et al., N. Engl. J. Med., 382: 1894-1905 (2020)). For example, one study showed that treatment of HCC in NASH mice using immune checkpoint inhibitor led to the production of more exhausted T cell phenotypes within the liver and overall disease progression (Pfister et al., Nature, 592(7854):450-456 (2021)). The same study performed a meta-analysis of three randomized phase III clinical trials that tested inhibitors of PD-L1 or PD-1 in more than 1,600 patients with advanced HCC and revealed that immune therapy did not improve survival in patients with non-viral HCC (Pfister et al., Nature, 592(7854):450-456 (2021)). Additional strategies need to be implemented to increase survival for HCC patients especially those with underlying fatty liver / NASH disease.

[0009] SUMMARY

[0010] This document provides methods and materials for treating a mammal (e.g., a human) having cancer (e.g., a cancer including one or more solid tumors). For example, this document provides methods and materials for using (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide to treat a mammal (e.g., a human) having cancer. In some cases, a mammal (e.g., a human such as a human having cancer) can be administered (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide to sensitize the cancer to ICB and, optionally, can also be administered one or more immune checkpoint inhibitors to treat the cancer. In some cases, a mammal (e.g., a human such as a human having cancer) can be administered (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide to enhance cellular immunotherapy (also referred to as adoptive cell therapy) and, optionally, can also be administered one or more cellular immunotherapies (e.g., engineered or expanded TILs) to treat the cancer.

[0011] As demonstrated herein, dual inhibition of NOS2 and COX2 can improve the efficacy of immune checkpoint inhibitors. Having the ability to improve the efficacy of immune checkpoint inhibitors as described herein (e.g., by administering (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide) provides a unique and unrealized opportunity to improve the treatment outcomes for (e.g., the survival of) mammals (e.g., humans) having cancer such as HCC.

[0012] In general, one aspect of this document features methods for treating a mammal having cancer where the methods can include, or consist essentially of, (a) administering an inhibitor of aNOS2 polypeptide to the mammal; (b) administering an inhibitor of a COX2 polypeptide to the mammal; and (c) administering an immune checkpoint inhibitor or a cellular immunotherapy to the mammal. The mammal can be a human. The cancer can include a solid tumor. The cancer can be a liver cancer, a breast cancer, a pancreatic cancer, an ovarian cancer, a lung cancer, a glioblastoma, a melanoma, a colorectal cancer, a brain cancer, an endocrine cancer, a sarcoma, or a testicular cancer. The inhibitor of the NOS2 polypeptide can be 1400W, GW274150, cindunistat (SD-6010), L-NMMA, or BYK 191023. The inhibitor of the COX2 polypeptide can be celecoxib, etoricoxib, meloxicam, diclofenac, or 2-acetoxybenzoic acid. The method can include administering the immune checkpoint inhibitor to the mammal, where the immune checkpoint inhibitor can be pembrolizumab, nivolumab, cemiplimab, ipilimumab, tremelimumab, durvalumab, dostarlimab, avelumab, atezolizumab, relatlimab, spartalizumab, camrelizumab, sintilimab, tislelizumab, toripalimab, BMS-8, BMS-37, BMS-202, BMS-230, BMS-242, BMS-1001, BMS-1166, SB415286, vorinostat, decitabine, entitostat, JQ1, BET151, GSK503, panobinostat, ACY-241, azacytidine, DB36, DB71, DB15, CVN, MGCD0103, SNDX-275, IMP32, BMS986016, TSR-022, Sym023, ATIK2a, DZNep, AMP-224, AMP-514, KN035, CK-301, AUNP12, CAPO, BMS-986189, NewEl, or DX11. In some cases, the inhibitor of the NOS2 polypeptide can be 1400W, the COX2 polypeptide can be celecoxib, and the immune checkpoint inhibitor can be atezolizumab, durvalumab, or avelumab. The method can include administering the cellular immunotherapy to the mammal, where the cellular immunotherapy can be administration of autologous T cells, administration of allogenic T cells, a chimeric antigen receptor (CAR) T cell therapy, a tumor-infiltrating lymphocyte (TIL) therapy, a natural killer (NK) cell therapy, administration of a dendritic cell (DC) vaccine, a TCR-engineered T cell therapy, or a macrophage therapy.

[0013] In another aspect, this document features methods for sensitizing a cancer within a mammal to an immune checkpoint inhibitor where the methods can include, or consist essentially of, (a) administering an inhibitor of a NOS2 polypeptide to the mammal; and (b) administering an inhibitor of a COX2 polypeptide to the mammal. The mammal can be a human. The cancer can include a solid tumor. The cancer can be a liver cancer, a breast cancer, a pancreatic cancer, an ovarian cancer, a lung cancer, a glioblastoma, a melanoma, a colorectal cancer, a brain cancer, an endocrine cancer, a sarcoma, or a testicular cancer. The inhibitor of the NOS2 polypeptide can be 1400W, GW274150, cindunistat (SD-6010), L- NMMA, or BYK 191023. The inhibitor of the COX2 polypeptide can be celecoxib, etoricoxib, meloxicam, diclofenac, or 2-acetoxybenzoic acid. The immune checkpoint inhibitor can be pembrolizumab, nivolumab, cemiplimab, ipilimumab, tremelimumab, durvalumab, dostarlimab, avelumab, atezolizumab, relatlimab, spartalizumab, camrelizumab, sintilimab, tislelizumab, toripalimab, BMS-8, BMS-37, BMS-202, BMS-230, BMS-242, BMS-1001, BMS-1166, SB415286, vorinostat, decitabine, entitostat, JQ1, BET151, GSK5O3, panobinostat, ACY-241, azacytidine, DB36, DB71, DB15, CVN, MGCD0103, SNDX-275, IMP32, BMS986016, TSR-022, Sym023, ATIK2a, DZNep, AMP-224, AMP- 514, KN035, CK-301, AUNP12, CA-170, BMS-986189, NewEl, or DX11. The method also can include administering the immune checkpoint inhibitor to the mammal.

[0014] In another aspect, this document features methods for enhancing cellular immunotherapy within a mammal having cancer where the methods can include, or consist essentially of, (a) administering an inhibitor of a NOS2 polypeptide to the mammal; and (b) administering an inhibitor of a COX2 polypeptide to the mammal. The mammal can be a human. The cancer can include a solid tumor. The cancer can be a liver cancer, a breast cancer, a pancreatic cancer, an ovarian cancer, a lung cancer, a glioblastoma, a melanoma, a colorectal cancer, a brain cancer, an endocrine cancer, a sarcoma, or a testicular cancer. The inhibitor of the NOS2 polypeptide can be 1400W, GW274150, cindunistat (SD-6010), L- NMMA, or BYK 191023. The inhibitor of the COX2 polypeptide can be celecoxib, etoricoxib, meloxicam, diclofenac, or 2-acetoxybenzoic acid. The cellular immunotherapy can be administration of autologous T cells, administration of allogenic T cells, a CAR T cell therapy, a TIL therapy, a NK cell therapy, administration of a DC vaccine, a TCR-engineered T cell therapy, or a macrophage therapy. The method can include administering the cellular immunotherapy to the mammal. The method can include administering an immune checkpoint inhibitor to the mammal. The immune checkpoint inhibitor can be pembrolizumab, nivolumab, cemiplimab, ipilimumab, tremelimumab, durvalumab, dostarlimab, avelumab, atezolizumab, relatlimab, spartalizumab, camrelizumab, sintilimab, tislelizumab, toripalimab, BMS-8, BMS-37, BMS-202, BMS-230, BMS-242, BMS-1001, BMS-1166, SB415286, vorinostat, decitabine, entitostat, JQ1, BET151, GSK503, panobinostat, ACY-241, azacytidine, DB36, DB71, DB15, CVN, MGCD0103, SNDX-275, IMP32, BMS986016, TSR-022, Sym023, ATIK2a, DZNep, AMP-224, AMP-514, KN035, CK-301, AUNP12, CA-170, BMS-986189, NewEl, or DXll.

[0015] In another aspect, this document features methods for treating a mammal having cancer where the methods can include, or consist essentially of, (a) administering 1400W to the mammal; (b) administering celecoxib to the mammal; and (c) administering an immune checkpoint inhibitor to the mammal where the immune checkpoint inhibitor can beatezolizumab, durvalumab, or avelumab. The mammal can be a human. The cancer can include a solid tumor. The cancer can be a liver cancer, a breast cancer, a pancreatic cancer, an ovarian cancer, a lung cancer, a glioblastoma, a melanoma, a colorectal cancer, a brain cancer, an endocrine cancer, a sarcoma, or a testicular cancer.

[0016] In another aspect, this document features methods for treating a mammal having cancer where the methods can include, or consist essentially of, (a) administering 1400W to the mammal; (b) administering celecoxib to the mammal; and (c) administering a TIL therapy to the mammal. The mammal can be a human. The cancer can include a solid tumor. The cancer can be a liver cancer, a breast cancer, a pancreatic cancer, an ovarian cancer, a lung cancer, a glioblastoma, a melanoma, a colorectal cancer, a brain cancer, an endocrine cancer, a sarcoma, or a testicular cancer. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Although methods and materials similar or equivalent to those described herein can be used to practice the invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

[0017] The details of one or more embodiments of the invention are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims.

[0018] BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figures 1A-1J. Molecular analysis of human hepatocellular carcinoma (HCC) tumors. Figures 1A and IB: Staining of human HCC tumors with HE, Masson and triple staining for CD8, iNOS and C0X2, respectively. Figure 1C: Analysis of NOS2, C0X2, and CD8 expression in tumors. Figure ID: Correlation of % positive expression of NOS2 and C0X2; inverse correlation of NOS2 and CD8. Figure IE: Distribution of staining in the tumor microenvironment (TME). Figure IF: Comparison of NOS2, C0X2, and CD8 in tumor vs. stroma. C0X2 is high in tumor, CD8 is high in stroma. Figure 1G: Recurrence-free survival (RFS) analysis in patients with high vs. low COX2 or NOS2. Figure 1H: Overall survival (OS) analysis comparing high vs. low C0X2 and NOS2 levels. Figure II: RFS and OS comparison between COX2lo"7NOS2lo" / CD8ll|8|' and COX2llls|7NOS2l"=l7CD8l°" groups. Figure 1J: Multivariate analysis of OS and RFS with various variables.

[0020] Figure 2: Kaplan-Meier survival analysis of subject analyzed in Figure 1.

[0021] Figures 3A-3G. Murine Tumor Growth and Immune Response. Figure 3A: Tumor model. Figure 3B: Macroscopic and microscopic tumor growth in treated mice. Figure 3C: Concentration Nitrite in liver and plasma following treatment. Figure 3D: Concentration of PGE2 in liver and plasma following treatment Figure 3E: NOS2 and C0X2 positive cells in TME. Figure 3F: CD8+and CD4+T cell comparison in the TME. Higher in triple double therapy. Figure 3G: B cells higher in double therapy.

[0022] Figures 4A-4G. PD-L1 Treatment in Mice. Figure 4A: Tumor size in mice treated with triple therapy. Figure 4B: Bulk RNA sequencing of triple-treated mice. Figure 4C: Analysis of gene expression changes. Figure 4D: Alterations in DNA damage checkpoints, DNA strand elongation, and cell cycle regulation. Figure 4E: Expression of protumor cytokines like G-CSF, M-CSF, IL-6, and TEMPI. Figure 4F: Expression of antitumor cytokines. Figure 4G: Survival comparison in lean mice with different treatments.

[0023] Figure 5. Additional single-cell analysis of lymphocytes and HALO image analysis.

[0024] Figure 6A-F. Single-cell analysis in mice. Analysis showing upregulation of activated T cells carrying CD226 protein.

[0025] Figures 7A-7E. Obese Mice Study. Figure 7A: Introduction. Figure 7B: Tumor growth differences in obese mice. Figure 7C: Expression of CD4, CD8, CD226 in TME. Figure 7D: Flow cytometry and RNA sequencing analysis of CD112 expression in tumor cells. Figure 7E: Survival analysis in obese mice including different immune checkpoint inhibitors.

[0026] Figures 8A-8E. T cell response to PGE2 and nitric oxide. Figure 8A: in vitro experiments of CD4 or CD8 T cells treated with PGE2 and various PGE2 inhibitors. Figure 8B: CD4 and CD8 T cells treated with nitric oxide and expression levels of Tigit. Figures 8C and 8D: PCR analysis of various genes and their expression levels in combinations of T cells and cancers cell following treatment with different doses of PGE2 or NO. Figure 8E: PVRL2 expression levels in cancer cells treated with different doses of PGE2 or NO.

[0027] Figures 9A-9F. Analysis of a human dataset. Figure 9A: Single-cell analysis in human cancers. Figure 9B: Expression levels of T cell markers. Figure 9C: Pathway GO analysis of T cells. Figure 9D: Expression levels of T cells and immunosuppressive or activating markers based on T cell subgroup. Figure 9E: Survival analysis based on the expression levels of EOMES, TBX21, and CD226. Figure 9F: Genes of interest correlating to survival shown in Figure 9E.

[0028] Figures 10A-10E. Treatment in mice. Figure 10A: Kaplan-Meier survival curves of mice (10-15 per group) inoculated with tumors and treated seven days later in designated groups. Mice were monitored daily by a trained technician. Survival comparison between triple therapy (anti-PD-Ll + GW274150 + celecoxib) and (anti-PD-Ll + 1400W + celecoxib) shows a significant difference (p < 0.0001) compared to control or anti-PD-Ll monotherapy. Figure 10B: Kaplan-Meier survival curves and liver images of mice in an experimental setup similar to that of Figure 10A, with the addition of anti-TIGIT therapy. Treatment was discontinued at day 175 post-tumor inoculation. Two mice in the quadruple therapy group remained tumor-free at day 271. Figure 10C: Kaplan-Meier survival curves and liver images showing a comparison of triple therapy (anti-PD-Ll + 1400W / GW274150) with either diclofenac or celecoxib (n=5 mice per group). Figure 10D: Triple therapy with anti-PD-1 instead of anti-PD-Ll (n=5 mice per group). Figure 10E: Similar treatment scheme to that of Figure 10D but using anti-CTLA-4 immunotherapy.

[0029] Figures 11 A-l ID. Figure 11 A: Two-day-old B6 mice were treated with STZ and received either anti-PD-L l monotherapy or triple therapy (anti-PD-L l + GW274150 + Celecoxib) for 20 weeks. Figure 1 IB: Mice euthanized and macroscopic tumor burden in the liver was assessed. Figure 11C: Mice underwent tail vein injection of RIL175 cells and were monitored for four weeks before receiving treatment as outlined in the figure. Figure 1 ID: At eight weeks, mice were euthanized, and lung tissues were harvested for both microscopic and macroscopic analysis.

[0030] Figures 12A-12C. Figure 12A: Schematic representation of tumor inoculation and treatment schedule, with corresponding macroscopic and microscopic images of tumors. Figures 12B-12C: HALO immunofluorescence analysis of cell subpopulations in the tumor microenvironment (TME) compared to a healthy liver background (n=5 mice per group).

[0031] Figures 13A-13F. Figure 13A: qPCR analysis of splenic T cells isolated from healthy mice and treated with varying doses of PGE2 in addition to CD3 / CD28 stimulation. Data are presented as fold change in gene expression relative to control. Figure 13B: qPCR analysis of T cells treated with nitric oxide (NO). Figure 13C: Flow cytometric analysis and gating for % positive cells following treatment with PGE2. Figure 13D: Similar analysis as performed in Figure 13C, but with NO treatment. Figure 13E: Pathway analysis using PKA agonists and antagonists, followed by flow cytometric gating. Figure 13F: JAK pathway analysis using flow cytometry. Figure 14. Mice treated as per schematic shown in Figure 10A, but with the addition of cindunistat in the triple therapy combination.

[0032] Figure 15. Surface nitrate measurements of macrophage cells following treatment with lipopolysaccharide (LPS) (positive control) and varying doses of known NOS inhibitors.

[0033] DETAILED DESCRIPTION

[0034] This document provides methods and materials for treating a mammal (e.g., a human) having cancer (e.g., a cancer including one or more solid tumors). For example, this document provides methods and materials for using (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide to treat a mammal (e.g., a human) having cancer. In some cases, administering (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide can be effective to sensitize a cancer to ICB (e.g., to administration of one or more immune checkpoint inhibitors). For example, a mammal (e.g., a human such as a human having cancer) can be administered (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide to sensitize the cancer to ICB and, optionally, can also be administered one or more immune checkpoint inhibitors to treat the cancer. In some cases, administering (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide can be effective to enhance cellular immunotherapy (e.g., administration of one or more immunotherapies). For example, a mammal (e.g., a human such as a human having cancer) can be administered (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide to enhance cellular immunotherapy and, optionally, can also be administered one or more cellular immunotherapies to treat the cancer.

[0035] Any appropriate mammal having cancer can be treated as described herein (e.g., by administering (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide). Examples of mammals that can have cancer and can be treated as described herein include, without limitation, humans, non-human primates (e.g., monkeys), horses, bovine species, porcine species, dogs, cats, mice, and rats. In some cases, a human having cancer can be treated by administering (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide.

[0036] A mammal (e.g., a human) having any type of cancer can be treated as described herein (e.g., by administering (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide). In some cases, a cancer that can be treated as described herein can include one or more solid tumors. In some cases, a cancer that can be treated as described herein can be a blood cancer. In some cases, a cancer treated as described herein can be a primary cancer. In some cases, a cancer treated as described herein can be a metastatic cancer. In some cases, a cancer treated as described herein can be a refractory cancer. In some cases, a cancer treated as described herein can have failed multiple lines of chemotherapy and / or immunotherapy. Examples of cancers that can be treated as described herein include, without limitation, liver cancers (e.g., hepatocellular cancers such as hepatocellular carcinomas (HCCs) and cholangiocarcinomas), breast cancers (e.g., triplenegative breast cancers), pancreatic cancers, ovarian cancers, lung cancers (e.g., small cell lung cancers and non-small cell lung cancers such as lung adenocarcinomas), glioblastomas, melanomas, colorectal cancers, brain cancers (e.g., meningiomas, schwannomas, and astrocytomas), endocrine cancers (e.g., papillary thyroid cancers, follicular thyroid cancers, and anaplastic thyroid cancers), sarcomas, and testicular cancers. In some cases, a cancer treated as described herein (e.g., by administering (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide) can be a metastatic cancer (e.g., a metastatic colorectal cancer, a metastatic breast cancer, and a metastatic pancreatic cancer) that has spread to the liver. In some cases, a cancer that can be treated as described herein (e.g., by administering (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide) can be a brain cancer (e.g., a meningioma, a schwannoma, or an astrocytoma), an endocrine cancer (e.g., a papillary thyroid cancer, a follicular thyroid cancer, or an anaplastic thyroid cancer), or a sarcoma. In some cases, a cancer that can be treated as described herein (e.g., by administering (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide) can be a brain cancer (e.g., a meningioma, a schwannoma, or an astrocytoma), an endocrine cancer (e.g., a papillary thyroid cancer, a follicular thyroid cancer, or an anaplastic thyroid cancer), a sarcoma, or a testicular cancer. In some cases, a cancer that can be treated as described herein (e.g., by administering (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide) can be a glioblastoma, a melanoma, a HCC, a metastatic colorectal cancer, or a metastatic ovarian cancer.

[0037] In some cases, the methods described herein also can include identifying a mammal as having cancer. Examples of methods that can be used to identify a mammal as having cancer include, without limitation, physical examination, laboratory tests (e.g., blood and / or urine), biopsy, imaging tests (e.g., X-ray, PET / CT, MRI, and / or ultrasound), nuclear medicine scans (e.g., bone scans), endoscopy, and / or genetic tests.

[0038] Once identified as having cancer, a mammal can be treated as described herein (e.g., by administering (a) one or more inhibitors of aNOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide).

[0039] A mammal (e.g., a human) having cancer can be administered or instructed to selfadminister any appropriate one or more (e.g., one, two, three, four, or more) inhibitors of a NOS2 polypeptide. In some cases, a mammal (e.g., a human) having cancer can be administered or instructed to self-administer two or more inhibitors of a NOS2 polypeptide.

[0040] An inhibitor of a NOS2 polypeptide can be an inhibitor of NOS2 polypeptide activity (e.g., anti-NOS2 antibodies such as neutralizing anti-NOS2 antibodies and small molecules that target a NOS2 polypeptide) or an inhibitor of NOS2 polypeptide expression (e.g., nucleic acid molecules designed to induce RNA interference (RNAi) of NOS2 polypeptide expression such as antisense oligonucleotides (ASOs), siRNA molecules, and shRNA molecules).

[0041] An inhibitor of a NOS2 polypeptide can inhibit any appropriate NOS2 polypeptide. In some cases, a NOS2 polypeptide that can be inhibited by an inhibitor of a NOS2 polypeptide that can be used to treat a mammal (e.g., a human) having cancer as described herein can be an inducible NOS (iNOS) polypeptide. Examples of NOS2 polypeptides that can be inhibited by an inhibitor of a NOS2 polypeptide that can be used to treat a mammal (e.g., a human) having cancer as described herein include, without limitation, those set forth in National Center for Biotechnology Information (NCBI) accession no. NP 000616.

[0042] Examples of inhibitors of a NOS2 polypeptide that can be used to treat a mammal (e.g., a human) having cancer as described herein (e.g., by administering (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide) can be as shown in Table 1.

[0043] Table 1. Inhibitors of NOS2 polypeptides.

[0044] In some cases, an inhibitor of a NOS2 polypeptide that can be used to treat a mammal (e.g., a human) having cancer as described herein can be as described elsewhere (see, e.g., Ho et al., Cephalalgia, 30(12): 1458-67 (2010); le Graverand et al., Ann. Rheum. Dis., 2013 Feb;72(2): 187-95 (2013); Chung et al., Sei. Transl. Med., 13(624):eabj5070 (2021); Vannini et al., Redox. Biol., 6:334-343 (2015); Minhas et al., Med. Res. Rev., 40(3):823-855 (2020); and Granados-Principal et al., Breast Cancer Research 17:25 (2015)).

[0045] A mammal (e.g., a human) having cancer can be administered or instructed to selfadminister any appropriate one or more (e.g., one, two, three, four, or more) inhibitors of a COX2 polypeptide. In some cases, a mammal (e.g., a human) having cancer can be administered or instructed to self-administer two or more inhibitors of a COX2 polypeptide.

[0046] An inhibitor of a COX2 polypeptide can be an inhibitor of COX2 polypeptide activity (e.g., anti-COX2 antibodies such as neutralizing anti-COX2 antibodies and small molecules that target a COX2 polypeptide) or an inhibitor of COX2 polypeptide expression (e.g., nucleic acid molecules designed to induce RNAi of NOS2 polypeptide expression such as ASOs, siRNA molecules, and shRNA molecules).

[0047] An inhibitor of a COX2 polypeptide can inhibit any appropriate COX2 polypeptide. Examples of COX2 polypeptides that can be inhibited by an inhibitor of a C0X2 polypeptide that can be used to treat a mammal (e.g., a human) having cancer as described herein include, without limitation, those set forth in NCBI accession no. NM_000963 or NP_000954.1.

[0048] Examples of inhibitors of a C0X2 polypeptide that can be used to treat a mammal (e.g., a human) having cancer as described herein (e.g., by administering (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a C0X2 polypeptide) can be as shown in Table 2.

[0049] Table 2. Inhibitors of COX2 polypeptides.

[0050] In some cases, an inhibitor of a C0X2 polypeptide that can be used to treat a mammal (e.g., a human) having cancer as described herein can be as described elsewhere (see, e.g., Hashemi Goradel et al., J. Cell. Physiol., 234(5): 5683 -5699 (2019); and Pu et al., Front. Oncol., 11 :637504 (2021)).

[0051] In some cases, administering (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide can be effective to sensitize a cancer to ICB (e.g., to administration of one or more immune checkpoint inhibitors). In some cases, a mammal (e.g., a human such as a human having cancer) can be administered (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide to sensitize the cancer to ICB and, optionally, can also be administered one or more immune checkpoint inhibitors to treat the cancer. An immune checkpoint inhibitor can inhibit one or more polypeptides involved in an immune checkpoint pathway. Examples of immune checkpoint pathways include, without limitation, PD-1 / PD-L1 pathways, PD-1 / PD-L2 pathways, CTLA-4 pathways, TRAIL pathways, LAG-3 pathways, TIM-3 pathways, BTLA pathways, TIGIT pathways, VISTA pathways, OX40 / OX40L pathways, CD27 / CD70 pathways, ICOS / ICOSL pathways, and CD226 pathways. An immune checkpoint inhibitor can inhibit any polypeptide involved in an immune checkpoint pathway. Examples of polypeptides involved in an immune checkpoint pathway that can be inhibited by an immune checkpoint inhibitor as described herein include, without limitation, PD-1 polypeptides, PD-L1 polypeptides, CTL4A polypeptides, LAG-3 polypeptides, TIM-3 polypeptides, B7-H3 polypeptides, B7-H4 polypeptides, TIGIT polypeptides, A2aR polypeptides, CD73 polypeptides, NKG2A polypeptides, PVRIG polypeptides, PVRL2 polypeptides, HAVCR2 polypeptides, IDO1 polypeptides, SIGLEC polypeptides, KIR polypeptides, and LILRB polypeptides.

[0052] An immune checkpoint inhibitor can inhibit polypeptide activity of a polypeptide involved in an immune checkpoint pathway or can inhibit polypeptide expression of a polypeptide involved in an immune checkpoint pathway. Examples of compounds that can inhibit polypeptide activity of a polypeptide involved in an immune checkpoint pathway include, without limitation, antibodies (e.g., neutralizing antibodies) that target (e.g., target and bind) to a polypeptide involved in an immune checkpoint pathway and small molecules that target (e.g., target and bind) to a polypeptide involved in an immune checkpoint pathway. Examples of compounds that can inhibit polypeptide expression of a polypeptide involved in an immune checkpoint pathway include, without limitation, nucleic acid molecules designed to induce RNA interference of polypeptide expression of a polypeptide involved in an immune checkpoint pathway (e.g., a siRNA molecule or a shRNA molecule), antisense molecules that can target (e.g., are complementary to) nucleic acid encoding a polypeptide involved in an immune checkpoint pathway, and miRNAs that can target (e.g., are complementary to) nucleic acid encoding a polypeptide involved in an immune checkpoint pathway.

[0053] In some cases, an immune checkpoint inhibitor can be an antibody (e.g., a monoclonal antibody). For example, an immune checkpoint inhibitor can be an anti-PD-1 antibody, an anti-PD-Ll antibody, an anti-CTL4A antibody, an anti -LAG-3 antibody, an anti-TIM-3 antibody, an anti-B7-H3 antibody, an anti-B7-H4 antibody, an anti-TIGIT antibody, an anti-A2aR antibody, an anti-CD73 antibody, an anti-NKG2A antibody, an anti- PVRIG antibody, an anti-PVRL2 antibody, an anti-CD27 antibody, an anti-CD40 antibody, an anti-OX40 antibody, an anti-GITR antibody, an anti-HAVCR2 antibody, or an anti-IDOl antibody. In some cases, an immune checkpoint inhibitor can be a bispecific antibody such as a bispecific T cell engager (BiTE).

[0054] Examples of immune checkpoint inhibitors that can be used as described herein include, without limitation, those set forth in Table 3.

[0055] Table 3. Exemplary immune checkpoint inhibitors

[0056]

[0057] In some cases, an immune checkpoint inhibitor can be as described elsewhere (see, e g., Smith et al., Am. J. Transl. Res., 11 (2):529-541 (2019) at, for example, Table 1 ; and Terranova-Barberio et al., Immunotherapy, 8(6):705-719 (2016) at, for example, Table 1). In some cases, a mammal (e.g., a human) having cancer can be administered 1400W and celecoxib (e.g., CELEBREX®). For example, 1400W and celecoxib (e.g., CELEBREX®) can be administered to a mammal (e.g., a human) having cancer (e.g., a cancer that demonstrate little to no responsiveness to ICB) to sensitize the cancer to ICB. In some cases, a mammal (e.g., a human) having cancer can be administered 1400W and celecoxib (e.g., CELEBREX®) to sensitize the cancer to an anti-PD-Ll antibody (e.g., atezolizumab, durvalumab, or avelumab), and, optionally, can be administered the anti-PD-Ll antibody to treat the cancer.

[0058] In some cases, a mammal (e.g., a human) having cancer can be administered 1400W and celecoxib (e.g., CELEBREX®). For example, 1400W and celecoxib (e.g., CELEBREX®) can be administered to a mammal (e.g., a human) having cancer (e.g., a cancer that demonstrate little to no responsiveness to ICB) to sensitize the cancer to ICB. In some cases, a mammal (e.g., a human) having cancer can be administered 1400W and celecoxib (e.g., CELEBREX®) to sensitize the cancer to an anti-PD-1 antibody (e.g., pembrolizumab, nivolumab, or cemiplimab), and, optionally, can be administered the anti-PD-Ll antibody to treat the cancer.

[0059] In some cases, a mammal (e.g., a human) having cancer can be administered (a) one or more inhibitors of a NOS2 polypeptide, (b) one or more inhibitors of a C0X2 polypeptide, and (c) one or more immune checkpoint inhibitors to treat the cancer. Exemplary combinations for treating cancer are set forth in Tables 4-6.

[0060] Table 4. Exemplary combinations

[0061]

[0062] Table 5. Exemplary combinations Table 6. Exemplary combinations

[0063] In some cases, administering (a) one or more inhibitors of a N0S2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide can be effective to enhance one or more cellular immunotherapies. In some cases, a mammal (e.g., a human such as a human having cancer) can be administered (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide to enhance cellular immunotherapy and, optionally, can also be administered one or more cellular immunotherapies to treat the cancer.

[0064] A cellular immunotherapy can be any appropriate type of cellular immunotherapy. In some cases, a cellular immunotherapy can enhance effectiveness of administration of immune cells. In some cases, a cellular immunotherapy can include engineered immune cells. In some cases, a cellular immunotherapy can include expanded immune cells. Examples of cellular immunotherapies include, without limitation, administration of autologous T cells, administration of allogenic T cells, chimeric antigen receptor (CAR) T cell therapies, TIL therapies, natural killer (NK) cell therapies, administration of a dendritic cell (DC) vaccines, TCR-engineered T cell therapies, and macrophage therapies. In some cases, administering (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide can be effective to enhance a CAR T cell therapy. In some cases, administering (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide can be effective to enhance a TIL therapy.

[0065] In some cases, a cellular immunotherapy can be as described elsewhere (see, e.g., Hayes, Ir. J. Med. Set., 190(l):41 -57 (2021); and Finck et al., Nat. Med., 28(4):678-689).

[0066] In some cases, a mammal (e.g., a human) having cancer can be administered 1400W and celecoxib (e g., CELEBREX®). For example, 1400W and celecoxib (e.g., CELEBREX®) can be administered to a mammal (e.g., a human) having cancer (e.g., a cancer that demonstrate little to no responsiveness to cellular immunotherapy) to enhance cellular immunotherapy. In some cases, a mammal (e.g., a human) having cancer can be administered 1400W and celecoxib (e.g., CELEBREX®) to enhance one or more cellular immunotherapies, and, optionally, can be administered a TIL therapy to treat the cancer. In some cases, a mammal (e.g., a human) having cancer can be administered (a) one or more inhibitors of a NOS2 polypeptide, (b) one or more inhibitors of a C0X2 polypeptide, and (c) one or more cellular immunotherapies to treat the cancer. Exemplary combinations for treating cancer are set forth in Table 7.

[0067] Table 7. Exemplary combinations

[0068] In some cases, a mammal (e.g., a human) having cancer can be administered (a) one or more inhibitors of a NOS2 polypeptide, (b) one or more inhibitors of a COX2 polypeptide, (c) one or more immune checkpoint inhibitors, and (d) one or more cellular immunotherapies to treat the cancer. For example, any of the exemplary combinations for treating cancer set forth in Tables 4-6 can be designed to include one or more cellular immunotherapies included in Table 7 (e.g., one or more of TIL therapy, CAR T cell therapy, NK cell therapy, TCR engineered T cell therapy, dendritic cell vaccine therapy, and macrophage therapy).

[0069] In some cases, one or more inhibitors of a NOS2 polypeptide and one or more inhibitors of a COX2 polypeptide (and, optionally, one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies) can be formulated into a composition (e.g., a pharmaceutically acceptable composition) for administration to a mammal (e.g., a human) having cancer. For example, a composition can include (a) one or more inhibitors of a NOS2 polypeptide and / or (b) one or more inhibitors of a COX2 polypeptide. For example, one or more inhibitors of a NOS2 polypeptide and / or one or more inhibitors of a COX2 polypeptide can be formulated together with one or more pharmaceutically acceptable carriers (additives), excipients, and / or diluents. In some cases, a pharmaceutically acceptable carrier, excipient, or diluent can be a naturally occurring pharmaceutically acceptable carrier, excipient, or diluent. In some cases, a pharmaceutically acceptable carrier, excipient, or diluent can be a non-naturally occurring (e g., an artificial or synthetic) pharmaceutically acceptable carrier, excipient, or diluent. Examples of pharmaceutically acceptable carriers, excipients, and diluents that can be used in a composition described herein (e.g., a pharmaceutically acceptable composition) include, without limitation, serum proteins (e g., human serum albumin), water, and salts or electrolytes (e.g., phosphate salts, saline, protamine sulfate, and DMSO).

[0070] In some cases, one or more inhibitors of a NOS2 polypeptide and one or more inhibitors of a COX2 polypeptide (and, optionally, one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies) can be administered to a mammal at the same time (e.g., in a single composition).

[0071] One or more inhibitors of a NOS2 polypeptide and / or one or more inhibitors of a COX2 polypeptide (and, optionally, one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies) can be administered to a mammal by any appropriate route. For example, a composition including (a) one or more inhibitors of a NOS2 polypeptide and / or (b) one or more inhibitors of a COX2 polypeptide can be administered locally or systemically.

[0072] In some cases, one or more inhibitors of a NOS2 polypeptide and / or one or more inhibitors of a COX2 polypeptide (and, optionally, one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies) can be designed for oral administration. Compositions suitable for oral administration include, without limitation, solid compositions, liquid compositions, and gel-like compositions. In some cases, a composition including (a) one or more inhibitors of a NOS2 polypeptide and / or (b) one or more inhibitors of a COX2 polypeptide can be designed for parenteral (e.g., subcutaneous, intramuscular, intravenous, intraperitoneal, and intratumoral) administration. Compositions suitable for parenteral administration include, without limitation, aqueous and non-aqueous sterile injection solutions that can contain anti-oxidants, buffers, bacteriostats, and solutes which render the composition isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents.

[0073] In some cases, a composition including (a) one or more inhibitors of a NOS2 polypeptide and / or (b) one or more inhibitors of a COX2 polypeptide (and, optionally, one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies) can be presented in unit-dose or multi-dose containers, for example, sealed ampules and vials, and may be stored in a freeze dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules, and tablets.

[0074] In some cases, one or more inhibitors of a NOS2 polypeptide and one or more inhibitors of a COX2 polypeptide (and, optionally, one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies) can be administered a mammal (e.g., a human) separately. For example, one or more inhibitors of a NOS2 polypeptide and one or more inhibitors of a COX2 polypeptide can be administered to a mammal at the same time (e g., concurrently) as independent compositions. When one or more inhibitors of a NOS2 polypeptide and one or more inhibitors of a COX2 polypeptide are administered concurrently, the composition including the one or more inhibitors of a NOS2 polypeptide and the composition including the one or more inhibitors of a COX2 polypeptide can be administered to a mammal within from about 1 second to about 15 minutes (e.g., about 2 seconds to about 15 minutes, about 5 seconds to about 15 minutes, about 10 seconds to about 15 minutes, about 15 seconds to about 15 minutes, about 1 second to about 10 minutes, about 1 second to about 5 minutes, or about 5 seconds to about 10 minutes) of each other.

[0075] In some cases, a composition including one or more inhibitors of a NOS2 polypeptide and a composition including one or more inhibitors of a COX2 polypeptide can be administered a mammal (e.g., a human) at different times. When a composition including one or more inhibitors of a NOS2 polypeptide and a composition including one or more inhibitors of a COX2 polypeptide are administered at different times, the composition including the one or more inhibitors of a NOS2 polypeptide and the composition including the one or more inhibitors of a COX2 polypeptide can be administered to a mammal with from about 16 minutes to about 48 hours (e.g., about 16 minutes to about 45 hours, about 16 minutes to about 36 hours, about 16 minutes to about 24 hours, about 16 minutes to about 12 hours, about 16 minutes to about 8 hours, about 16 minutes to about 6 hours, about 16 minutes to about 4 hours, about 30 minutes to about 48 hours, about 1 hour to about 48 hours, about 2 hours to about 48 hours, about 4 hours to about 48 hours, about 6 hours to about 48 hours, or 8 hours minutes to about 48 hours) between each administration.

[0076] When one or more inhibitors of a NOS2 polypeptide and one or more inhibitors of a COX2 polypeptide (and, optionally, one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies) are administered as separate compositions (e.g., administered concurrently as separate compositions or administered as separate compositions with from about 16 minutes to about 48 hours between each administration), each composition can be administered to a mammal by any appropriate route. In some cases, a composition including one or more inhibitors of a NOS2 polypeptide and a composition including one or more inhibitors of a COX2 polypeptide can be administered by the same route. In some cases, a composition including one or more inhibitors of a NOS2 polypeptide and a composition including one or more inhibitors of a COX2 polypeptide can be administered by different routes.

[0077] When one or more inhibitors of a NOS2 polypeptide and one or more inhibitors of a COX2 polypeptide (and, optionally, one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies) are administered as separate compositions (e.g., administered concurrently as separate compositions or administered as separate compositions with from about 0 seconds to about 15 minutes between each administration), a composition including the one or more inhibitors of a NOS2 polypeptide can be administered first, and a composition including the one or more inhibitors of a COX2 polypeptide administered second, or vice versa. One or more inhibitors of aNOS2 polypeptide can be administered to a mammal (e.g., a human) having cancer in any appropriate amount (e.g., any appropriate dose). In some cases, an effective dose of one or more inhibitors of a NOS2 polypeptide can be a flat dose. In some cases, an effective amount of cindunistat can be from about 50 mg to about 200 mg (e.g., from about 50 mg to about 185 mg, about 50 mg to about 170 mg, about 50 mg to about 155 mg, about 50 mg to about 140 mg, about 50 mg to about 125 mg, about 50 mg to about 110 mg, about 50 mg to about 95 mg, about 50 mg to about 80 mg, about 50 mg to about 65 mg, about 65 mg to about 200 mg, about 80 mg to about 200 mg, about 95 mg to about 200 mg, about 110 mg to about 200 mg, about 125 mg to about 200 mg, about 140 mg to about 200 mg, about 155 mg to about 200 mg, about 170 mg to about 200 mg, about 185 mg to about 200 mg, about 65 mg to about 185 mg, about 80 mg to about 170 mg, about 95 mg to about 155 mg, about 110 mg to about 140 mg, about 50 mg to about 80 mg, about 65 mg to about 95 mg, about 80 mg to about 110 mg, about 95 mg to about 125 mg, about 110 mg to about 140 mg, about 125 mg to about 155 mg, about 140 mg to about 170 mg, about 155 mg to about 185 mg, or about 170 mg to about 200 mg) (e.g., per day). In some cases, an effective amount of GW-274150 can be from about 60 mg to about 120 mg (e.g., from about 60 mg to about 115 mg, about 60 mg to about 110 mg, about 60 mg to about 105 mg, about 60 mg to about 100 mg, about 60 mg to about 95 mg, about 60 mg to about 90 mg, about 60 mg to about 85 mg, about 60 mg to about 80 mg, about 60 mg to about 75 mg, about 60 mg to about 70 mg, about 60 mg to about 65 mg, about 65 mg to about 120 mg, about 70 mg to about 120 mg, about 75 mg to about 120 mg, about 80 mg to about 120 mg, about 85 mg to about 120 mg, about 90 mg to about 120 mg, about 95 mg to about 120 mg, about 100 mg to about 120 mg, about 105 mg to about 120 mg, about 110 mg to about 120 mg, about 115 mg to about 120 mg, about 65 mg to about 115 mg, about 70 mg to about 110 mg, about 75 mg to about 105 mg, about 80 mg to about 100 mg, about 85 mg to about 95 mg, about 60 mg to about 70 mg, about 65 mg to about 75 mg, about 70 mg to about 80 mg, about 75 mg to about 85 mg, about 80 mg to about 90 mg, about 85 mg to about 95 mg, about 90 mg to about 100 mg, about 95 mg to about 105 mg, about 100 mg to about 110 mg, about 105 mg to about 115 mg, or about 110 mg to about 120 mg) (e.g., per day). In some cases, an effective dose of one or more inhibitors of a NOS2 polypeptide can be based on the body weight of a mammal (e.g., a human) to be treated as described herein. In some cases, an effective amount of GW274150 can be from about 0.1 mg / kg to about 200 mg / kg (e.g., from about 0.1 mg / kg to about 180 mg / kg, about 0.1 mg / kg to about 160 mg / kg, about 0.1 mg / kg to about 140 mg / kg, about 0.1 mg / kg to about 120 mg / kg, about 0.1 mg / kg to about 100 mg / kg, about 0.1 mg / kg to about 80 mg / kg, about 0.1 mg / kg to about 60 mg / kg, about 0.1 mg / kg to about 40 mg / kg, about 0.1 mg / kg to about 20 mg / kg, about 0.1 mg / kg to about 10 mg / kg, about 0.1 mg / kg to about 1 mg / kg, about 0.1 mg / kg to about 0.5 mg / kg, about 0.5 mg / kg to about 200 mg / kg, about 1 mg / kg to about 200 mg / kg, about 10 mg / kg to about 200 mg / kg, about 20 mg / kg to about 200 mg / kg, about 40 mg / kg to about 200 mg / kg, about 60 mg / kg to about 200 mg / kg, about 80 mg / kg to about 200 mg / kg, about 100 mg / kg to about 200 mg / kg, about 120 mg / kg to about 200 mg / kg, about 140 mg / kg to about 200 mg / kg, about 160 mg / kg to about 200 mg / kg, about 180 mg / kg to about 200 mg / kg, about 0.5 mg / kg to about 180 mg / kg, about 1 mg / kg to about 160 mg / kg, about 10 mg / kg to about 140 mg / kg, about 20 mg / kg to about 120 mg / kg, about 40 mg / kg to about 100 mg / kg, about 60 mg / kg to about 8 mg / kg, about 60 mg / kg, or about 90 mg / kg) (e.g., per day). In some cases, an effective amount of L-NMMA can be from about 5 mg / kg to about 20 mg / kg (e.g., from about 5 mg / kg to about 18 mg / kg, about 5 mg / kg to about 16 mg / kg, about 5 mg / kg to about 14 mg / kg, about 5 mg / kg to about 12 mg / kg, about 5 mg / kg to about 10 mg / kg, about 5 mg / kg to about 9 mg / kg, about 5 mg / kg to about 7 mg / kg, about 7 mg / kg to about 20 mg / kg, about 9 mg / kg to about 20 mg / kg, about 10 mg / kg to about 20 mg / kg, about 12 mg / kg to about 20 mg / kg, about 14 mg / kg to about 20 mg / kg, about 16 mg / kg to about 20 mg / kg, about 18 mg / kg to about 20 mg / kg, about 7 mg / kg to about 18 mg / kg, about 9 mg / kg to about 16 mg / kg, about 10 mg / kg to about 14 mg / kg, about 5 mg / kg to about 9 mg / kg, about 7 mg / kg to about 10 mg / kg, about 9 mg / kg to about 12 mg / kg, about 10 mg / kg to about 14 mg / kg, about 12 mg / kg to about 16 mg / kg, about 14 mg / kg to about 18 mg / kg, about 16 mg / kg to about 20 mg / kg, about 5 mg / kg, about 7.5 mg / kg, about 10 mg / kg, about 12.5 mg / kg, about 15 mg / kg, about 17.5 mg / kg, or about 20 mg / kg) (e.g., per day). In some cases, an effective amount of 1400W can be from about 0.3 mg / kg to about 60 mg / kg (e.g., from about 0.3 mg / kg to about 40 mg / kg, about 0.3 mg / kg to about 20 mg / kg, about 0.3 mg / kg to about 10 mg / kg, about 0.3 mg / kg to about 5 mg / kg, about 0.3 mg / kg to about 1 mg / kg, about 0.3 mg / kg to about 0.5 mg / kg, about 0.5 mg / kg to about 60 mg / kg, about 1 mg / kg to about 60 mg / kg, about 5 mg / kg to about 60 mg / kg, about 10 mg / kg to about 60 mg / kg, about 20 mg / kg to about 60 mg / kg, about 40 mg / kg to about 60 mg / kg, about 0.5 mg / kg to about 40 mg / kg, about 1 mg / kg to about 20 mg / kg, or about 5 mg / kg to about 10 mg / kg) (e.g., per day). In some cases, an effective dose of one or more inhibitors of a NOS2 polypeptide can be as described elsewhere (see, e.g., Hellio le Graverand et al., Ann. Rheum. Dis., 72(2): 187-95 (2013); Aiderton et al., Br. J. Pharmacol., 145(3):301 - 12 (2005); Dugo et al., Br. J. Pharmacol., 141(6):979-87 (2004); Seymour et al., Clin. Exp. Rheumatol., 30(2):254-61 (2012); Singh et al., Am. J. Respir. Crit. Care Med., 176(10):988-93 (2007); and Chung et al., Sci. Transl. Med., 13(624):eabj5070 (2021)). The effective amount of one or more inhibitors of a NOS2 polypeptide can remain constant or can be adjusted as a sliding scale or variable dose depending on the mammal’s response to treatment. Various factors can influence the actual effective amount used for a particular application. For example, the frequency of administration, duration of treatment, use of multiple treatment agents, route of administration, and / or severity of the cancer in the mammal being treated may require an increase or decrease in the actual effective amount administered.

[0078] One or more inhibitors of aNOS2 polypeptide can be administered to a mammal (e.g., a human) having cancer at any appropriate frequency. The frequency of administration can be any frequency that can treat a mammal having cancer without producing significant toxicity to the mammal. For example, the frequency of administration can be from about twice a day to about one every other day, from about once a day to about once a week, from about once a day to about once a month, from about once a week to about once a month, or from about twice a month to about once a month. The frequency of administration can remain constant or can be variable during the duration of treatment. As with the effective amount, various factors can influence the actual frequency of administration used for a particular application. For example, the effective amount, duration of treatment, use of multiple treatment agents, and / or route of administration may require an increase or decrease in administration frequency.

[0079] One or more inhibitors of a NOS2 polypeptide can be administered to a mammal (e.g., a human) having cancer for any appropriate duration. An effective duration can be any duration that can treat a mammal having cancer without producing significant toxicity to the mammal. For example, the effective duration can vary from several weeks to several months, from several months to several years, or from several years to a lifetime. Multiple factors can influence the actual effective duration used for a particular treatment. For example, an effective duration can vary with the frequency of administration, effective amount, use of multiple treatment agents, and / or route of administration.

[0080] One or more inhibitors of a COX2 polypeptide can be administered to a mammal (e.g., a human) having cancer in any appropriate amount (e.g., any appropriate dose). In some cases, an effective dose of one or more inhibitors of a COX2 polypeptide can be a flat dose. In some cases, an effective dose of one or more inhibitors of a COX2 polypeptide can be based on the body of a mammal (e.g., a human) to be treated as described herein. In some cases, an effective amount of celecoxib can be from about 100 mg to about 4800 mg (e.g., from about 100 mg to about 4000 mg, about 100 mg to about 3200 mg, about 100 mg to about 2400 mg, about 100 mg to about 1600 mg, about 100 mg to about 800 mg, about 100 mg to about 660 mg, about 100 mg to about 520 mg, about 100 mg to about 380 mg, about 100 mg to about 240 mg, about 240 mg to about 4800 mg, about 380 mg to about 4800 mg, about 520 mg to about 4800 mg, about 660 mg to about 4800 mg, about 800 mg to about 4800 mg, about 1600 mg to about 4800 mg, about 2400 mg to about 4800 mg, about 3200 mg to about 4800 mg, about 4000 mg to about 4800 mg, about 240 mg to about 4000 mg, about 380 mg to about 3200 mg, about 520 mg to about 2400 mg, about 660 mg to about 1600 mg, about 100 mg to about 380 mg, about 240 mg to about 520 mg, about 380 mg to about 660 mg, about 520 mg to about 800 mg, about 660 mg to about 1600 mg, about 800 mg to about 2400 mg, about 1600 mg to about 3200 mg, about 2400 mg to about 4000 mg, or about 3200 mg to about 4800 mg) (e.g., per day). In some cases, an effective amount of etoricoxib can be from about 30 mg to about 120 mg (e.g., from about 30 mg to about 110 mg, about 30 mg to about 100 mg, about 30 mg to about 90 mg, about 30 mg to about 80 mg, about 30 mg to about 70 mg, about 30 mg to about 55 mg, about 30 mg to about 45 mg, about 45 mg to about 120 mg, about 55 mg to about 120 mg, about 70 mg to about 120 mg, about 80 mg to about 120 mg, about 90 mg to about 120 mg, about 100 mg to about 120 mg, about 110 mg to about 120 mg, about 45 mg to about 110 mg, about 55 mg to about 100 mg, about 70 mg to about 90 mg, about 30 mg to about 55 mg, about 45 mg to about 70 mg, about 55 mg to about 80 mg, about 70 mg to about 90 mg, about 80 mg to about 100 mg, about 90 mg to about 110 mg, or about 100 mg to about 120 mg) (e.g., per day). In some cases, an effective amount of meloxicam can be from about 7.5 mg to about 15 mg (e.g., from about 7.5 mg to about 14.0 mg, about 7.5 mg to about 13.0 mg, about 7.5 mg to about 12.0 mg, about 7.5 mg to about 11.0 mg, about 7.5 mg to about 10.0 mg, about 7.5 mg to about 9.5 mg, about 7.5 mg to about 9.0 mg, about 7.5 mg to about 8.5 mg, about 7.5 mg to about 8.0 mg, about 8.0 mg to about 15.0 mg, about 8.5 mg to about 15.0 mg, about 9.0 mg to about 15.0 mg, about 9.5 mg to about 15.0 mg, about 10.0 mg to about 15.0 mg, about 11.0 mg to about 15.0 mg, about 12.0 mg to about 15.0 mg, about 13.0 mg to about 15.0 mg, about 14.0 mg to about 15.0 mg, about 8.0 mg to about 14.0 mg, about 8.5 mg to about 13.0 mg, about 9.0 mg to about 12.0 mg, about 9.5 mg to about 11.0 mg, about 7.5 mg to about 8.5 mg, about 8.0 mg to about 9.0 mg, about 8.5 mg to about 9.5 mg, about 9.0 mg to about 10.0 mg, about 9.5 mg to about 11.0 mg, about 10.0 mg to about 12.0 mg, about 11.0 mg to about 13.0 mg, about 12.0 mg to about 14.0 mg, or about 13.0 mg to about 15.0 mg) (e.g., per day). In some cases, an effective amount of diclofenac can be from about 50 mg to about 150 mg (e.g., from about 50 mg to about 140 mg, about 50 mg to about 130 mg, about 50 mg to about 120 mg, about 50 mg to about 110 mg, about 50 mg to about 100 mg, about 50 mg to about 90 mg, about 50 mg to about 80 mg, about 50 mg to about 70 mg, about 50 mg to about 60 mg, about 60 mg to about 150 mg, about 70 mg to about 150 mg, about 80 mg to about 150 mg, about 90 mg to about 150 mg, about 100 mg to about 150 mg, about 110 mg to about 150 mg, about 120 mg to about 150 mg, about 130 mg to about 150 mg, about 140 mg to about 150 mg, about 60 mg to about 140 mg, about 70 mg to about 130 mg, about 80 mg to about 120 mg, about 90 mg to about 110 mg, about 50 mg to about 70 mg, about 60 mg to about 80 mg, about 70 mg to about 90 mg, about 80 mg to about 100 mg, about 90 mg to about 110 mg, about 100 mg to about 120 mg, about 110 mg to about 130 mg, about 120 mg to about 140 mg, or about 130 mg to about 150 mg) (e.g., per day). In some cases, an effective amount of 2-acetoxybenzoic acid can be from about 75 mg to about 4000 mg (e.g., from about 75 mg to about 3320 mg, about 75 mg to about 2640 mg, about 75 mg to about 1960 mg, about 75 mg to about 1280 mg, about 75 mg to about 600 mg, about 75 mg to about 495 mg, about 75 mg to about 390 mg, about 75 mg to about 285 mg, about 75 mg to about 180 mg, about 180 mg to about 4000 mg, about 285 mg to about 4000 mg, about 390 mg to about 4000 mg, about 495 mg to about 4000 mg, about 600 mg to about 4000 mg, about 1280 mg to about 4000 mg, about 1960 mg to about 4000 mg, about 2640 mg to about 4000 mg, about 3320 mg to about 4000 mg, about 180 mg to about 3320 mg, about 285 mg to about 2640 mg, about 390 mg to about 1960 mg, about 495 mg to about 1280 mg, about 75 mg to about 285 mg, about 180 mg to about 390 mg, about 285 mg to about 495 mg, about 390 mg to about 600 mg, about 495 mg to about 1280 mg, about 600 mg to about 1960 mg, about 1280 mg to about 2640 mg, about 1960 mg to about 3320 mg, or about 2640 mg to about 4000 mg) (per day). In some cases, an effective amount of nabumetone can be from about 500 mg to about 2000 mg (e.g., from about 500 mg to about 1800 mg, about 500 mg to about 1600 mg, about 500 mg to about 1400 mg, about 500 mg to about 1200 mg, about 500 mg to about 1020 mg, about 500 mg to about 860 mg, about 500 mg to about 680 mg, about 680 mg to about 2000 mg, about 860 mg to about 2000 mg, about 1020 mg to about 2000 mg, about 1200 mg to about 2000 mg, about 1400 mg to about 2000 mg, about 1600 mg to about 2000 mg, about 1800 mg to about 2000 mg, about 680 mg to about 1800 mg, about 860 mg to about 1600 mg, about 1020 mg to about 1400 mg, about 500 mg to about 860 mg, about 680 mg to about 1020 mg, about 860 mg to about 1200 mg, about 1020 mg to about 1400 mg, about 1200 mg to about 1600 mg, about 1400 mg to about 1800 mg, or about 1600 mg to about 2000 mg) (e.g., per day). In some cases, an effective amount of ibuprofen can be from about 200 mg to about 3200 mg (e.g., from about 200 mg to about 2800 mg, about 200 mg to about 2400 mg, about 200 mg to about 2000 mg, about 200 mg to about 1600 mg, about 200 mg to about 1200 mg, about 200 mg to about 1000 mg, about 200 mg to about 800 mg, about 200 mg to about 600 mg, about 200 mg to about 400 mg, about 400 mg to about 3200 mg, about 600 mg to about 3200 mg, about 800 mg to about 3200 mg, about 1000 mg to about 3200 mg, about 1200 mg to about 3200 mg, about 1600 mg to about 3200 mg, about 2000 mg to about 3200 mg, about 2400 mg to about 3200 mg, about 2800 mg to about 3200 mg, about 400 mg to about 2800 mg, about 600 mg to about 2400 mg, about 800 mg to about 2000 mg, about 1000 mg to about 1600 mg, about 200 mg to about 600 mg, about 400 mg to about 800 mg, about 600 mg to about 1000 mg, about 800 mg to about 1200 mg, about 1000 mg to about 1600 mg, about 1200 mg to about 2000 mg, about 1600 mg to about 2400 mg, about 2000 mg to about 2800 mg, or about 2400 mg to about 3200 mg) (e.g., per day). In some cases, an effective amount of naproxen can be from about 220 mg to about 1375 mg (e g., from about 220 mg to about 1260 mg, about 220 mg to about 1145 mg, about 220 mg to about 1030 mg, about 220 mg to about 915 mg, about 220 mg to about 800 mg, about 220 mg to about 680 mg, about 220 mg to about 560 mg, about 220 mg to about 460 mg, about 220 mg to about 340 mg, about 340 mg to about 1375 mg, about 460 mg to about 1375 mg, about 560 mg to about 1375 mg, about 680 mg to about 1375 mg, about 800 mg to about 1375 mg, about 915 mg to about 1375 mg, about 1030 mg to about 1375 mg, about 1145 mg to about 1375 mg, about 1260 mg to about 1375 mg, about 340 mg to about 1260 mg, about 460 mg to about 1145 mg, about 560 mg to about 1030 mg, about 680 mg to about 915 mg, about 220 mg to about 460 mg, about 340 mg to about 560 mg, about 460 mg to about 680 mg, about 560 mg to about 800 mg, about 680 mg to about 915 mg, about 800 mg to about 1030 mg, about 915 mg to about 1145 mg, about 1030 mg to about 1260 mg, or about 1145 mg to about 1375 mg) (per day). In some cases, an effective amount of sulindac can be from about 150 mg to about 400 mg (e.g., from about 150 mg to about 370 mg, about 150 mg to about 340 mg, about 150 mg to about 310 mg, about 150 mg to about 280 mg, about 150 mg to about 250 mg, about 150 mg to about 220 mg, about 150 mg to about 180 mg, about 180 mg to about 400 mg, about 220 mg to about 400 mg, about 250 mg to about 400 mg, about 280 mg to about 400 mg, about 310 mg to about 400 mg, about 340 mg to about 400 mg, about 370 mg to about 400 mg, about 180 mg to about 370 mg, about 220 mg to about 340 mg, about 250 mg to about 310 mg, about 150 mg to about 220 mg, about 180 mg to about 250 mg, about 220 mg to about 280 mg, about 250 mg to about 310 mg, about 280 mg to about 340 mg, about 310 mg to about 370 mg, or about 340 mg to about 400 mg) (e.g., per day). In some cases, an effective dose of one or more inhibitors of a COX2 polypeptide can be as described elsewhere (see, e g., Silverstein et al., JAMA, 284:1247-55 (2000); Steinbach et al., N. Engl. J. Med., 342: 1946-52 (2000); and Bertagnolli et al., N. Engl. J. Med., 355:873-84 (2006)). The effective amount of one or more inhibitors of a COX2 polypeptide can remain constant or can be adjusted as a sliding scale or variable dose depending on the mammal’s response to treatment. Various factors can influence the actual effective amount used for a particular application. For example, the frequency of administration, duration of treatment, use of multiple treatment agents, route of administration, and / or severity of the cancer in the mammal being treated may require an increase or decrease in the actual effective amount administered.

[0081] One or more inhibitors of a COX2 polypeptide can be administered to a mammal (e.g., a human) having cancer at any appropriate frequency. The frequency of administration can be any frequency that can treat a mammal having cancer without producing significant toxicity to the mammal. For example, the frequency of administration can be from about twice a day to about one every other day, from about once a day to about once a week, from about once a day to about once a month, from about once a week to about once a month, or from about twice a month to about once a month. The frequency of administration can remain constant or can be variable during the duration of treatment. As with the effective amount, various factors can influence the actual frequency of administration used for a particular application. For example, the effective amount, duration of treatment, use of multiple treatment agents, and / or route of administration may require an increase or decrease in administration frequency.

[0082] One or more inhibitors of a COX2 polypeptide can be administered to a mammal (e.g., a human) having cancer for any appropriate duration. An effective duration can be any duration that can treat a mammal having cancer without producing significant toxicity to the mammal. For example, the effective duration can vary from several weeks to several months, from several months to several years, or from several years to a lifetime. Multiple factors can influence the actual effective duration used for a particular treatment. For example, an effective duration can vary with the frequency of administration, effective amount, use of multiple treatment agents, and / or route of administration.

[0083] One or more immune checkpoint inhibitors can be administered to a mammal (e.g., a human) having cancer in any appropriate amount (e.g., any appropriate dose). In some cases, an effective dose of one or more immune checkpoint inhibitors can be a flat dose. In some cases, an effective amount of cemiplimab can be about 350 mg (e.g., about 100 mg, about 200 mg, about 500 mg, or about 1000 mg). In some cases, an effective amount of avelumab can be about 800 mg (e.g., about 600 mg, about 700 mg, about 900 mg, or about 1000 mg). In some cases, an effective amount of relatimab can be about 160 mg (e.g., about 100 mg, about 140 mg, about 180 mg, or about 200 mg). In some cases, an effective amount of relatlimab + nivolumab can be about 480 mg (e.g., about 100 mg, about 200 mg, about 600 mg, or about 800 mg). In some cases, an effective amount of sintilimab can be about 200 mg (e.g., about 100 mg, about 150 mg, about 250 mg, or about 400 mg). In some cases, an effective amount of camrelizuma can be about 200 mg (e.g., about 100 mg, about 150 mg, about 250 mg, or about 400 mg). In some cases, an effective amount of toripalimab can be about 240 mg (e.g., about 100 mg, about 200 mg, about 300 mg, or about 400 mg). In some cases, an effective amount of tislelizumab can be about 200 mg (e.g., about 100 mg, about 150 mg, about 250 mg, or about 400 mg). In some cases, an effective amount of pembrolizumab can be from about 200 mg to about 400 mg (e.g., from about 200 mg to about 380 mg, about 200 mg to about 360 mg, about 200 mg to about 340 mg, about 200 mg to about 320 mg, about 200 mg to about 300 mg, about 200 mg to about 280 mg, about 200 mg to about 260 mg, about 200 mg to about 240 mg, about 200 mg to about 220 mg, about 220 mg to about 400 mg, about 240 mg to about 400 mg, about 260 mg to about 400 mg, about 280 mg to about 400 mg, about 300 mg to about 400 mg, about 320 mg to about 400 mg, about 340 mg to about 400 mg, about 360 mg to about 400 mg, about 380 mg to about 400 mg, about 220 mg to about 380 mg, about 240 mg to about 360 mg, about 260 mg to about 340 mg, about 280 mg to about 320 mg, about 200 mg to about 240 mg, about 220 mg to about 260 mg, about 240 mg to about 280 mg, about 260 mg to about 300 mg, about 280 mg to about 320 mg, about 300 mg to about 340 mg, about 320 mg to about 360 mg, about 340 mg to about 380 mg, or about 360 mg to about 400 mg). In some cases, an effective amount of nivolumab can be from about 240 mg to about 480 mg (e.g., from about 240 mg to about 460 mg, about 240 mg to about 440 mg, about 240 mg to about 420 mg, about 240 mg to about 400 mg, about 240 mg to about 380 mg, about 240 mg to about 360 mg, about 240 mg to about 340 mg, about 240 mg to about 320 mg, about 240 mg to about 300 mg, about 240 mg to about 280 mg, about 240 mg to about 260 mg, about 260 mg to about 480 mg, about 280 mg to about 480 mg, about 300 mg to about 480 mg, about 320 mg to about 480 mg, about 340 mg to about 480 mg, about 360 mg to about 480 mg, about 380 mg to about 480 mg, about 400 mg to about 480 mg, about 420 mg to about 480 mg, about 440 mg to about 480 mg, about 460 mg to about 480 mg, about 260 mg to about 460 mg, about 280 mg to about 440 mg, about 300 mg to about 420 mg, about 320 mg to about 400 mg, about 340 mg to about 380 mg, about 240 mg to about 280 mg, about 260 mg to about 300 mg, about 280 mg to about 320 mg, about 300 mg to about 340 mg, about 320 mg to about 360 mg, about 340 mg to about 380 mg, about 360 mg to about 400 mg, about 380 mg to about 420 mg, about 400 mg to about 440 mg, about 420 mg to about 460 mg, or about 440 mg to about 480 mg). In some cases, an effective amount of tremelimuma can be from about 75 mg to about 300 mg (e.g., from about 75 mg to about 275 mg, about 75 mg to about 250 mg, about 75 mg to about 225 mg, about 75 mg to about 200 mg, about 75 mg to about 175 mg, about 75 mg to about 155 mg, about 75 mg to about 135 mg, about 75 mg to about 115 mg, about 75 mg to about 95 mg, about 95 mg to about 300 mg, about 115 mg to about 300 mg, about 135 mg to about 300 mg, about 155 mg to about 300 mg, about 175 mg to about 300 mg, about 200 mg to about 300 mg, about 225 mg to about 300 mg, about 250 mg to about 300 mg, about 275 mg to about 300 mg, about 95 mg to about 275 mg, about 115 mg to about 250 mg, about 135 mg to about 225 mg, about 155 mg to about 200 mg, about 75 mg to about 115 mg, about 95 mg to about 135 mg, about 115 mg to about 155 mg, about 135 mg to about 175 mg, about 155 mg to about 200 mg, about 175 mg to about 225 mg, about 200 mg to about 250 mg, about 225 mg to about 275 mg, or about 250 mg to about 300 mg). In some cases, an effective amount of durvalumab can be from about 1200 mg to about 1500 mg (e.g., from about 1200 mg to about 1475 mg, about 1200 mg to about 1450 mg, about 1200 mg to about 1425 mg, about 1200 mg to about 1400 mg, about 1200 mg to about 1375 mg, about 1200 mg to about 1350 mg, about 1200 mg to about 1325 mg, about 1200 mg to about 1300 mg, about 1200 mg to about 1275 mg, about 1200 mg to about 1250 mg, about 1200 mg to about 1225 mg, about 1225 mg to about 1500 mg, about 1250 mg to about 1500 mg, about 1275 mg to about 1500 mg, about 1300 mg to about 1500 mg, about 1325 mg to about 1500 mg, about 1350 mg to about 1500 mg, about 1375 mg to about 1500 mg, about 1400 mg to about 1500 mg, about 1425 mg to about 1500 mg, about 1450 mg to about 1500 mg, about 1475 mg to about 1500 mg, about 1225 mg to about 1475 mg, about 1250 mg to about 1450 mg, about 1275 mg to about 1425 mg, about 1300 mg to about 1400 mg, about 1325 mg to about 1375 mg, about 1200 mg to about 1250 mg, about 1225 mg to about 1275 mg, about 1250 mg to about 1300 mg, about 1275 mg to about 1325 mg, about 1300 mg to about 1350 mg, about 1325 mg to about 1375 mg, about 1350 mg to about 1400 mg, about 1375 mg to about 1425 mg, about 1400 mg to about 1450 mg, about 1425 mg to about 1475 mg, or about 1450 mg to about 1500 mg). In some cases, an effective amount of dostarlimab can be from about 500 mg to about 1000 mg (e.g., from about 500 mg to about 950 mg, about 500 mg to about 900 mg, about 500 mg to about 850 mg, about 500 mg to about 800 mg, about 500 mg to about 750 mg, about 500 mg to about 700 mg, about 500 mg to about 650 mg, about 500 mg to about 600 mg, about 500 mg to about 550 mg, about 550 mg to about 1000 mg, about 600 mg to about 1000 mg, about 650 mg to about 1000 mg, about 700 mg to about 1000 mg, about 750 mg to about 1000 mg, about 800 mg to about 1000 mg, about 850 mg to about 1000 mg, about 900 mg to about 1000 mg, about 950 mg to about 1000 mg, about 550 mg to about 950 mg, about 600 mg to about 900 mg, about 650 mg to about 850 mg, about 700 mg to about 800 mg, about 500 mg to about 600 mg, about 550 mg to about 650 mg, about 600 mg to about 700 mg, about 650 mg to about 750 mg, about 700 mg to about 800 mg, about 750 mg to about 850 mg, about 800 mg to about 900 mg, about 850 mg to about 950 mg, or about 900 mg to about 1000 mg). In some cases, an effective amount of atezolizuma can be from about 840 mg to about 1680 mg (e.g., from about 840 mg to about 1600 mg, about 840 mg to about 1520 mg, about 840 mg to about 1440 mg, about 840 mg to about 1360 mg, about 840 mg to about 1280 mg, about 840 mg to about 1200 mg, about 840 mg to about 1140 mg, about 840 mg to about 1080 mg, about 840 mg to about 1020 mg, about 840 mg to about 960 mg, about 840 mg to about 900 mg, about 900 mg to about 1680 mg, about 960 mg to about 1680 mg, about 1020 mg to about 1680 mg, about 1080 mg to about 1680 mg, about 1140 mg to about 1680 mg, about 1200 mg to about 1680 mg, about 1280 mg to about 1680 mg, about 1360 mg to about 1680 mg, about 1440 mg to about 1680 mg, about 1520 mg to about 1680 mg, about 1600 mg to about 1680 mg, about 900 mg to about 1600 mg, about 960 mg to about 1520 mg, about 1020 mg to about 1440 mg, about 1080 mg to about 1360 mg, about 1140 mg to about 1280 mg, about 840 mg to about 960 mg, about 900 mg to about 1020 mg, about 960 mg to about 1080 mg, about 1020 mg to about 1140 mg, about 1080 mg to about 1200 mg, about 1140 mg to about 1280 mg, about 1200 mg to about 1360 mg, about 1280 mg to about 1440 mg, about 1360 mg to about 1520 mg, about 1440 mg to about 1600 mg, or about 1520 mg to about 1680 mg). In some cases, an effective dose of one or more immune checkpoint inhibitors can be based on the body weight of a mammal (e.g., a human) to be treated as described herein. In some cases, an effective amount of nivolumab can be about 3 mg / kg (e.g., about 1 mg / kg, about 2 mg / kg, about 5 mg / kg, or about 10 mg / kg). In some cases, an effective amount of durvalumab can be about 10 mg / kg (e.g., about 1 mg / kg, about 5 mg / kg, about 15 mg / kg, or about 20 mg / kg). In some cases, an effective amount of ipilimumab can be from about 1 mg / kg to about 3 mg / kg (e.g., from about 1 .0 mg / kg to about 2.8 mg / kg, about 1 .0 mg / kg to about 2.6 mg / kg, about 1.0 mg / kg to about 2.4 mg / kg, about 1.0 mg / kg to about 2.2 mg / kg, about 1.0 mg / kg to about 2.0 mg / kg, about 1.0 mg / kg to about 1.8 mg / kg, about 1.0 mg / kg to about 1.6 mg / kg, about 1.0 mg / kg to about 1.4 mg / kg, about 1.0 mg / kg to about 1.2 mg / kg, about 1.2 mg / kg to about 3.0 mg / kg, about 1.4 mg / kg to about 3.0 mg / kg, about 1.6 mg / kg to about 3.0 mg / kg, about

[0084] 1.8 mg / kg to about 3.0 mg / kg, about 2.0 mg / kg to about 3.0 mg / kg, about 2.2 mg / kg to about 3.0 mg / kg, about 2.4 mg / kg to about 3.0 mg / kg, about 2.6 mg / kg to about 3.0 mg / kg, about

[0085] 2.8 mg / kg to about 3.0 mg / kg, about 1.2 mg / kg to about 2.8 mg / kg, about 1.4 mg / kg to about 2.6 mg / kg, about 1.6 mg / kg to about 2.4 mg / kg, about 1.8 mg / kg to about 2.2 mg / kg, about 1.0 mg / kg to about 1.4 mg / kg, about 1.2 mg / kg to about 1.6 mg / kg, about 1.4 mg / kg to about

[0086] 1.8 mg / kg, about 1.6 mg / kg to about 2.0 mg / kg, about 1.8 mg / kg to about 2.2 mg / kg, about 2.0 mg / kg to about 2.4 mg / kg, about 2.2 mg / kg to about 2.6 mg / kg, about 2.4 mg / kg to about

[0087] 2.8 mg / kg, or about 2.6 mg / kg to about 3.0 mg / kg). The effective amount of one or more immune checkpoint inhibitors can remain constant or can be adjusted as a sliding scale or variable dose depending on the mammal’s response to treatment. Various factors can influence the actual effective amount used for a particular application. For example, the frequency of administration, duration of treatment, use of multiple treatment agents, route of administration, and / or severity of the cancer in the mammal being treated may require an increase or decrease in the actual effective amount administered.

[0088] One or more immune checkpoint inhibitors can be administered to a mammal (e.g., a human) having cancer at any appropriate frequency. The frequency of administration can be any frequency that can treat a mammal having cancer without producing significant toxicity to the mammal. For example, the frequency of administration can be from about twice a day to about one every other day, from about once a day to about once a week, from about once a day to about once a month, from about once a week to about once a month, or from about twice a month to about once a month. The frequency of administration can remain constant or can be variable during the duration of treatment. As with the effective amount, various factors can influence the actual frequency of administration used for a particular application. For example, the effective amount, duration of treatment, use of multiple treatment agents, and / or route of administration may require an increase or decrease in administration frequency.

[0089] One or more immune checkpoint inhibitors can be administered to a mammal (e.g., a human) having cancer for any appropriate duration. An effective duration can be any duration that can treat a mammal having cancer without producing significant toxicity to the mammal. For example, the effective duration can vary from several weeks to several months, from several months to several years, or from several years to a lifetime. Multiple factors can influence the actual effective duration used for a particular treatment. For example, an effective duration can vary with the frequency of administration, effective amount, use of multiple treatment agents, and / or route of administration.

[0090] In some cases, a dosing regimen for administering one or more checkpoint inhibitors to a mammal (e.g., a human) as described herein can be as shown in Table 8.

[0091] Table 8. Exemplary treatment regimens

[0092] One or more cellular immunotherapies can be administered to a mammal (e.g., a human) having cancer in any appropriate amount (e.g., any appropriate dose). In some cases, an effective amount of one or more cellular immunotherapies can include from about 1 billion cells to about 100 billion cells (e.g., per day) (e.g., from about 1 billion cells to about 70 billion cells, from about 1 billion cells to about 50 billion cells, from about 1 billion cells to about 30 billion cells, from about 1 billion cells to about 20 billion cells, from about 1 billion cells to about 10 billion cells, from about 20 billion cells to about 100 billion cells, from about 40 billion cells to about 100 billion cells, from about 60 billion cells to about 100 billion cells, from about 70 billion cells to about 100 billion cells, from about 80 billion cells to about 100 billion cells, from about 90 billion cells to about 100 billion cells, from about 10 billion cells to about 90 billion cells, from about 20 billion cells to about 80 billion cells, from about 30 billion cells to about 70 billion cells, from about 40 billion cells to about 60 billion cells, from about 10 billion cells to about 30 billion cells, from about 20 billion cells to about 40 billion cells, from about 30 billion cells to about 50 billion cells, from about 40 billion cells to about 60 billion cells, from about 50 billion cells to about 70 billion cells, from about 60 billion cells to about 80 billion cells, or from about 70 billion cells to about 90 billion cells (e.g., per day)). The effective amount of one or more cellular immunotherapies can remain constant or can be adjusted as a sliding scale or variable dose depending on the mammal’s response to treatment. Various factors can influence the actual effective amount used for a particular application. For example, the frequency of administration, duration of treatment, use of multiple treatment agents, route of administration, and / or severity of the cancer in the mammal being treated may require an increase or decrease in the actual effective amount administered.

[0093] One or more cellular immunotherapies can be administered to a mammal (e.g., a human) having cancer at any appropriate frequency. The frequency of administration can be any frequency that can treat a mammal having cancer without producing significant toxicity to the mammal. For example, the frequency of administration can be from about twice a day to about one every other day, from about once a day to about once a week, from about once a day to about once a month, from about once a week to about once a month, or from about twice a month to about once a month. The frequency of administration can remain constant or can be variable during the duration of treatment. As with the effective amount, various factors can influence the actual frequency of administration used for a particular application. For example, the effective amount, duration of treatment, use of multiple treatment agents, and / or route of administration may require an increase or decrease in administration frequency.

[0094] One or more immune cellular immunotherapies can be administered to a mammal (e.g., a human) having cancer for any appropriate duration. An effective duration can be any duration that can treat a mammal having cancer without producing significant toxicity to the mammal. For example, the effective duration can vary from several weeks to several months, from several months to several years, or from several years to a lifetime. Multiple factors can influence the actual effective duration used for a particular treatment. For example, an effective duration can vary with the frequency of administration, effective amount, use of multiple treatment agents, and / or route of administration.

[0095] In some cases, one or more inhibitors of a NOS2 polypeptide and one or more inhibitors of a COX2 polypeptide (and, optionally, one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies) can be administered to a mammal (e.g., a human) having cancer as the sole active agents to treat the cancer. In some cases, methods for treating a mammal (e.g., a human) as described herein (e.g., by administering (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide and, optionally, one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies) also can include administering to the mammal one or more (e.g., one, two, three, or more) additional agents used to treat cancer and / or performing one or more (e.g., one, two, three, or more) therapies used to treat cancer. For example, a combination therapy used to treat a mammal (e.g., a human) having cancer can include administering to the mammal (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide (and, optionally, administering one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies), and administering to the mammal one or more (e.g., one, two, three, or more) additional agents used to treat cancer. In some cases, an additional agent that can be administered to a mammal to treat cancer can be a chemotherapeutic agent. In some cases, an additional agent that can be administered to a mammal to treat cancer can be a cytotoxic agent. In some cases, an additional agent that can be administered to a mammal to treat cancer can be a targeted therapy. In some cases, an additional agent that can be administered to a mammal to treat cancer can be a hormone therapy. Examples of additional agents that can be administered to a mammal (e.g., a human) having cancer (e.g., a cancer including one or more solid tumors) to treat the mammal include, without limitation, sorafenib (e.g., NEXAVAR®), lenvatinib (e.g., LENVIMA®), goserelin (e.g., ZOLADEX®), leuprolide (e.g., ELIGARD® and LUPRON DEPOT®), bevacizumab (e.g., AVASTIN®), trastuzumab (e.g., HERCEPTIN®), axitinib (e.g., INLYTA®), cabozantinib (e.g., CABOMETYX®), olaparib (e.g., LYNPARZA®), palbociclib (e g., IB RANCE®), rituximab (e.g., RITUXAN®), and any combinations thereof. In cases where (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide (and, optionally, one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies) are used in combination with additional agents used to treat a mammal (e.g., a human) having cancer, the one or more additional agents can be administered at the same time (e.g., in a single composition containing (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide (and, optionally, one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies), and containing the one or more additional agents) or independently. For example, a composition including (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide (and, optionally, one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies) can be administered first, and the one or more additional agents administered second, or vice versa.

[0096] In some cases, a combination therapy used to treat a mammal (e.g., a human) having cancer can include administering to the mammal (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide (and, optionally, administering one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies), and can include performing one or more (e.g., one, two, three, or more) therapies used to treat cancer. Examples of additional therapies that can be used to treat a mammal (e.g., a human) having cancer include, without limitation, radiation therapies, and / or surgeries. In cases where (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide (and, optionally, one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies) are used in combination with one or more therapies used to treat a mammal (e.g., a human) having cancer, the one or more additional therapies can be performed at the same time or independently of the administration of the (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide (and, optionally, the one or more immune checkpoint inhibitors and / or the one or more cellular immunotherapies). For example, (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide (and, optionally, one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies) can be administered before, during, or after the one or more additional therapies are performed.

[0097] In some cases, the materials and methods provided herein can be used to reduce the number of immune cells present in a TME present within in a mammal (e.g., a human) having cancer. For example, a mammal having cancer (e.g., a human having cancer) in need of treatment thereof can be administered (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide (and, optionally, one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies) to reduce the number of immunosuppressive cells present in a TME present within the mammal. Examples of immunosuppressive cells that can be reduced in a TME present within a mammal (e.g., a human) having cancer as described herein (e.g., by administering (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide (and, optionally, one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies)) include, without limitation, regulatory T cells (Tregs), myeloid-derived suppressor cells (MDSCs), tumor-associated macrophages, cancer-associated fibroblasts, M2 macrophages, tumor-associated neutrophils, tumor-associated endothelial cells, regulatory stem cells, and cancer stem cells. Tn some cases, the methods and materials provided herein can be used as described herein to reduce the number of cancer cells in the mammal by, for example, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or more percent. In some cases, the methods and materials provided herein can be used as described herein to reduce the volume of one or more tumors in the mammal by, for example, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or more percent.

[0098] In some cases, the materials and methods provided herein can be used to increase the number of tumor infiltrating lymphocytes (TILs) present in a TME present within in a mammal (e.g., a human) having cancer. For example, a mammal having cancer (e.g., a human having cancer) in need of treatment thereof can be administered (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide (and, optionally, one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies) to increase the number of immunosuppressive cells present in a TME present within the mammal. In some cases, the methods and materials provided herein can be used as described herein to increase the number of TILs in a TME within the mammal by, for example, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or more percent.

[0099] In some cases, the materials and methods provided herein can be used to improve survival (e.g., recurrence-free survival (RFS) and / or overall survival (OS)) of a mammal (e.g., a human) having cancer. For example, a mammal having cancer (e.g., a human having cancer) in need of treatment thereof can be administered (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide (and, optionally, one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies) to improve survival of the mammal. For example, a mammal (e.g., a human) having cancer can be administered (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide, and can be administered (c) one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies to improve the survival of a mammal having cancer by, for example, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or more percent (e.g., as compared to a mammal having cancer that was administered only (a) and (b) or administered only (c)). For example, a mammal (e.g., a human) having cancer can be administered (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide, and can be administered (c) one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies to improve the survival of a mammal having cancer by, for example, at least 6 months (e.g., about 6 months, about 8 months, about 10 months, about 1 year, about 1.5 years, about 2 years, about 2.5 years, about 3 years, about 4 years, about 5 years, or more) (e.g., as compared to a mammal having cancer that was administered only (a) and (b) or administered only (c)).

[0100] In some cases, the materials and methods provided herein can be used to reduce the size of a cancer in a mammal (e.g., a human). For example, a mammal having cancer (e.g., a human having cancer) in need of treatment thereof can be administered (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide (and, optionally, one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies) to reduce the size of the cancer in the mammal. In some cases, a mammal (e.g., a human) having cancer can be administered (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide, and can be administered (c) one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies to reduce the number of cancer cells in the mammal by, for example, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or more percent (e.g., as compared to a mammal having cancer that was administered only (a) and (b) or administered only (c)). In some cases, a mammal (e.g., a human) having cancer can be administered (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide, and can be administered (c) one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies to reduce the volume of one or more tumors in the mammal by, for example, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or more percent (e.g., as compared to a mammal having cancer that was administered only (a) and (b) or administered only (c)).

[0101] In some cases, the materials and methods provided herein can include monitoring the mammal (e.g., the human) being treated as described herein (e.g., by administering (a) one or more inhibitors of a NOS2 polypeptide and (b) one or more inhibitors of a COX2 polypeptide (and, optionally, one or more immune checkpoint inhibitors and / or one or more cellular immunotherapies)). For example, the size of the cancer (e.g., the number of cancer cells and / or the volume of one or more tumors) present within a mammal can be monitored. Any appropriate method can be used to determine whether or not the size of the cancer present within a mammal is reduced. For example, imaging techniques can be used to assess the size of the cancer present within a mammal (e.g., a human).

[0102] The invention will be further described in the following examples, which do not limit the scope of the invention described in the claims.

[0103] EXAMPLES

[0104] Example 1: Inhibition of NOS and COX2 promotes T cell function and inhibits cancer growth

[0105] Hepatocellular carcinoma (HCC) is often unresectable when found and a rising cause of cancer death worldwide.

[0106] This Example demonstrates that co-inhibition of INOS and COX2 improved activation of T cells, slowed tumor growth, and improved survival of C57 / B6 mice bearing orthotopic transplants of HCC cells.

[0107] Methods

[0108] All mice received injection of 1 xlO6tumor cells into the liver parenchyma. Seven days later daily treatment with 1400W (iNOS inhibitor), celecoxib (COX2 inhibitor), 10F.9G2™ (an anti-PD-Ll antibody), or a combination of these agents was begun (double= 1400W+CELEBREX® ; tripl e= 1400W+CELEBREX®+anti-PD-L 1 ) . Sub sequent experiments were carried out with additional combinations of diclofenac (COX2 inhibitor), GW274150 (iNOS inhibitor), and a monoclonal anti-PD-1 antibody (RMP1-14) or a monoclonal anti-CTLA4 antibody (9D9). Treatment regimens and dosing were as provided in Table 9. Two weeks later the tumors were removed for flow cytometric examination of tumor and inflammatory cellular infdtrates and the cells were subjected to qPCR / bulk RNA sequencing and single cell analysis.

[0109] Table 9. Experimental treatment regimens

[0110] Results

[0111] All mice injected with RIL-175 developed macroscopic tumors by day 7 prior to start of treatment. Control untreated mice survived a median of 20 days, anti-PD-Ll treated mice survived a median of 35 days, double treated mice survived a median of 32 days, and mice that received triple therapy survived a median of 52 days (p<0.001). Each treatment alone increased the infiltration of non-exhausted CD4 and CD8 T cells into the tumor microenvironment (TME). The highest percentage of non-exhausted CD4 and CD8 T cells in the TME was found in mice treated with double or triple combination therapy (e g., as compared to control mice).

[0112] To determine the mechanism by which iNOS and COX2 inhibition led to reduced tumor growth in vivo (in addition to the PD-1 / PD-L1 system), single cell and bulk sequencing of the tumor infiltrating immune cells and the tumor cells, respectively, were performed. The combination of 1400W, CELEBREX®, and anti-PD-Ll showed a distinct population of highly activated T cells that expressed the marker CD226 (p< 10“86) compared to control or any other treatment combination. Bulk RNA sequencing of the tumor cells showed that CD112 had the highest expression in mice treated with the triple therapy. Thus, the degree of tumor growth reduction coincided with an activated T cell immune response, prolonged survival, and engagement of the emerging CD112 (tumor) with CD226 (T cell activation) pathway. Confirmatory experiments in vitro showed that treatment of RIL- 175 with PGE2 (end product of COX2 pathway) and / or NO led to several fold downregulation of both CD112 on cancers as well as CD226 on T cells.

[0113] Together, these results demonstrate that inhibition of NOS2 / COX2 led to increased infiltration of T cells, decreased tumor growth, and longer survival. Accordingly, a combination of one or more inhibitors of a NOS2 polypeptide and one or more inhibitors of a C0X2 polypeptide can be used to sensitize the cancer to ICB and, optionally, can be administered together with one or more immune checkpoint inhibitors to treat a mammal (e.g., a human) having cancer (e.g., a cancer including one or more solid tumors).

[0114] Example 2: N0S2 and C0X2 Expression in the Tumor Microenvironment and Therapeutic Outcomes in Hepatocellular Carcinoma

[0115] This Examples describes the interplay between NOS2 and COX2 expression in hepatocellular carcinoma (HCC) and demonstrates the impact of NOS2 and C0X2 expression on tumor progression and response to therapy in human and murine models. For example, the correlation of NOS2 and C0X2 with HCC progression was explored, focusing on recurrence-free survival (RFS) and overall survival (OS) in human subjects, and extended these findings to a murine model.

[0116] Methods

[0117] Human Study

[0118] Analysis of 141 HCC tumors for NOS2, C0X2, and CD8 expression.

[0119] Sample Collection and Preparation

[0120] Patient Selection and Consent: Patients diagnosed with HCC who were scheduled for surgical resection were selected for this study. Surgical Resection: Tumors were surgically resected, with care taken to minimize ischemia time. Immediately after resection, a portion of the tumor and adjacent healthy liver tissue were separated.

[0121] Tissue Processing: The collected tissue samples were fixed in 10% neutral buffered formalin for 24-48 hours at room temperature to preserve cellular details and prevent degradation.

[0122] Paraffin Embedding: After fixation, tissues were dehydrated through a graded series of ethanol, cleared in xylene, and embedded in paraffin blocks. This process preserved the tissue structure and allowed for long-term storage.

[0123] Sectioning: Paraffin-embedded tissues were sectioned at a thickness of 4 pm using a microtome. Sections were mounted on positively charged glass slides to enhance tissue adhesion.

[0124] Inmnmofluore scent Staining

[0125] Deparaffinization and Rehydration: Slides were placed in xylene to remove paraffin, then rehydrated through a graded alcohol series ending in distilled water.

[0126] Antigen Retrieval: Slides were heated in a citrate buffer (pH 6.0) or EDTA buffer (pH 8.0) for antigen retrieval. This step unmasked epitopes, improving antibody accessibility.

[0127] Blocking: A blocking solution (e.g., 5% BSA or normal serum) was applied to prevent non-specific binding of antibodies.

[0128] Primary Antibody Incubation: Slides were incubated with primary antibodies against NOS2, COX2, and CD8. Incubation was typically done overnight at 4°C.

[0129] Secondary Antibody Incubation: Apply fluorophore-conjugated secondary antibodies specific to the primary antibodies' host species. Incubate according to the manufacturer's recommendations, usually for 1-2 hours at room temperature in the dark.

[0130] Nuclear Staining: Nuclei were counterstained with a dye like DAPI to facilitate cell identification and analysis.

[0131] Mounting: An anti-fade mounting medium was applied to the slides to preserve fluorescence and prevent photobleaching. Image Analysis

[0132] Digital Scanning: Stained slides were scanned at a high resolution using a digital slide scanner to create digital images for analysis.

[0133] HALO Software Analysis: The HALO image analysis software (Indica Labs) was employed to quantify the expression and distribution of NOS2, C0X2, and CD8+ T cells within the tumor and adjacent healthy tissue. Specific analysis modules designed for IHC quantification were utilized.

[0134] Marker Quantification: The software was calibrated to recognize DAB staining for NOS2, C0X2, and CD8, and hematoxylin staining for nuclei. Parameters were adjusted to accurately segment and quantify positive staining in both tumor and healthy tissues.

[0135] Cell Distribution Analysis: The spatial distribution of CD8+ T cells in relation to NOS2 and C0X2 expressing cells was analyzed to understand the immune contexture of the tumor microenvironment.

[0136] Statistical Analysis

[0137] Data Compilation: Data on the percentage of positive cells, staining intensity, and spatial distribution were compiled for each sample.

[0138] Murine Study

[0139] Orthotopic HCC injections in C57BL / 6 mice, treated with celecoxib, 1400W, or GW274150 (GW).

[0140] Animal Preparation

[0141] 8-week-old C57BL / 6 mice were acclimated to the laboratory environment for at least 1 week before the experiment. Mice were housed under specific pathogen-free conditions, with ad libitum access to food and water, and maintain them on a 12-hour light / dark cycle.

[0142] Cell Preparation

[0143] RIL-175 HCC cells were cultivated under recommended conditions until they reached 80-90% confluency. Cells were harvested using trypsin-EDTA, washed with PBS, and resuspended in a suitable injection medium (e.g., PBS or Matrigel) at a concentration of 2 x 1OA6 cells / mL. The cell suspension was kept on ice until injection.

[0144] Anesthesia and Analgesia:

[0145] Mice were anesthetized using an appropriate method (e.g., isoflurane inhalation) to ensure sufficient depth of anesthesia throughout the procedure.

[0146] Analgesics were administered pre-operatively according to institutional guidelines to manage pain.

[0147] Surgical Procedure for Orthotopic Injection:

[0148] A laparotomy was performed through a small midline abdominal incision under sterile conditions.

[0149] The liver was gently exposed and 600,000 (300 pL) RIL-175 cells were injected directly into the left lateral lobe of the liver using a 27-gauge needle.

[0150] A few seconds after injection (to prevent leakage), the needle was withdrawn.

[0151] The abdominal wall was closed in two layers using absorbable sutures for the muscle layer and non-absorbable sutures or surgical staples for the skin.

[0152] Post-Operative Care

[0153] Mice were monitored closely for signs of pain, distress, or infection, and postoperative analgesia was administered as per the approved protocol.

[0154] Mice were returned to their housing once they were fully recovered from anesthesia.

[0155] Treatment Administration

[0156] Treatment Groups: Mice were divided into four groups: Control (vehicle), celecoxib, 1400W, and GW-treated groups.

[0157] Treatment Preparation and Administration: celecoxib, 1400W, and GW were prepared according to the manufacturer's instructions.

[0158] Treatments were administered orally (for celecoxib) or intraperitoneally (for 1400W and GW), depending on the compound's pharmacokinetics and bioavailability, 7 days after the HCC cell injection. Treatment regimens and dosing were as provided in Table 9. Treatment administration was continued for a specified duration as determined by the experimental design, and mice were monitored daily for health status and body weight.

[0159] Monitoring and Endpoints

[0160] Health Monitoring: Mice were observed daily for signs of tumor burden, distress, or adverse effects from the treatments. This included monitoring for weight loss, lethargy, hunched posture, and any other signs of illness.

[0161] Tumor Assessment: Depending on the study's endpoints, non-invasive imaging techniques (e.g., ultrasound or MRI) were used to monitor tumor growth and progression in vivo at predetermined intervals.

[0162] Euthanasia and Post-Mortem Analysis: Mice were euthanized humanely at the end of the study period or if they show signs of excessive tumor burden or distress.

[0163] Necropsies were performed to assess tumor growth and metastasis, liver and tumor tissues were collected for further histological and molecular analysis.

[0164] Results

[0165] Human tissue slides post-resection were used for a molecular analysis of human HCC tumors. Hematoxylin and Eosin (H&E) staining and Masson's trichrome staining (Figure 1A), along with immunofluorescent staining for CD8, NOS2, and COX2 markers (Figure IB- IF) were used to identify areas of cancer and distinguish between stroma and cancer areas. Recurrence-free survival (RFS) rates in patients having cancers either high or low expressions of C0X2 or NOS2 were determined (Figure G). Overall survival (OS) rates between patients having cancers with high or low expressions of C0X2 and N0S2 were compared (Figure 1H). RFS and OS outcomes for patients having cancers with varying expression levels of COX2, NOS2, and CD8 were also examined (Figure II) and multivariate analyses of OS and RFS were performed (Figure 1J). OS and RFS across various combinations of NOS2, COX2, and CD8 levels are shown in Figure 2. Patients with high NOS2 and / or C0X2 had significantly worse overall and recurrence-free survival compared to those with low expression levels (univariate and multivariate analysis). The majority of C0X2 was found within the tumor, whereas NOS2 was predominantly located within the stroma. A mouse model was used to evaluate tumor growth and immune response (Figure 3 A). Macroscopic and microscopic tumor burdens in animals treated with a vehicle control, CELEBREX®, 1400W, or a combination of CELEBREX® + 1400W were evaluated (Figure 3B). Plasma and liver expression levels of nitric oxide (Figure 3C) and PGE2 (Figure 3D) were examined in treated animals. Nitric oxide and PGE2 levels in response to various iNOS inhibitors were investigated. COX2 and NO levels within the tumor (Figure 3F), as well as expression levels of CD4, CD8, and B cells within the tumor (Figure 3G) were also assessed. Mice treated with either anti -iNOS (1400W), anti-COX2 (CELEBREX®), or a double combination exhibited significant reductions in tumor growth. They showed diminished levels of liver and plasma PGE2 (end product of the COX2 pathway) and nitrites (NO pathway end product). In addition, mice with double treatment had increased tumor infiltration of CD4, CD8, and B cells.

[0166] Tumor burdens in animals treated with different drugs were evaluated (Figure 4A), and PCR analysis of tumor cells in groups treated with vehicle, CELEBREX®, a combination of CELEBREX® + 1400W, or triple therapy (1400W, CELEBREX®, PD-L1) was performed for bulk RNA sequencing (Figure 4B). A heatmap of different expression levels of PVRL genes in shown in Figure 4C. A GSEA analysis of DNA pathways in mice treated with PD- L1 compared to those treated with CELEBREX® and 1400W is shown in Figure 4D. Protumor cytokines and chemokines in mice treated with various agents are shown in Figure 4E, with serum and / or plasma analyzed at the time of tumor harvest. Anti-tumor cytokines and chemokines in mice treated with various agents are shown in Figure 4F. Survival of mice treated with different drugs is plotted in Figure 4G. HALO tissue analysis was also performed for varying tissue markers in mice treated with different agents (Figure 5). Mice treated with the anti-PDLl inhibitor alone failed to control tumor growth. Mice treated with anti-PDLl therapy upregulated pro-tumor cytokines and chemokines. The addition of anti- iNOS and anti-COX2 inhibitors significantly reduced tumor burden, increased survival, and promoted anti-tumor cytokine production.

[0167] Single-cell analysis was performed in mice treated with various agents (Figure 6A). A heatmap of varying subgroups of the highest expressed genes is shown in Figure 6B. Genomic analysis of different T cell subgroups and their expression levels of CD8, CD4, and CD226 is shown in Figure 6C. Average expression levels of different T cell groups categorized into naive, cytotoxic, and activated are shown in Figure 6D. Gene expression pathway analysis of T cells in mice treated with triple therapy is shown in Figure 6E. A subgroup analysis of T cell coinhibitory, cytotoxic, costimulatory, or dysfunctional genes in various groups of treated mice is shown in Figure 6F. Single-cell analysis of mice undergoing treatment with anti-PDLl+1400W+ CELEBREX® compared to anti-PDLl shows an increase in T cell and B cell infiltration. These mice also had increased T cells expressing CD226 (to promote T cell survival and activation) and downregulated Tigit (to promote T cell dysfunction). These T cells were highly active and upregulated several pro-survival and anti-tumor cytokine pathways.

[0168] Obese mice (NASH) studies were also performed. Obese B6 BIO mice were injected with cancer, and treatment commenced after 7 days (Figure 7A). Macroscopic tumors of mice treated with vehicle control, 1400W, CELEBREX®, anti-PD-Ll, or a combination of 1400W, CELEBREX®, and anti-PD-Ll are shown in Figure 7B. Quantified percentage of CD8+ and CD4+, CD226+ and CD8+, or CD4+ Tigit+ T cells are shown in Figure 7C. Tumor cells were isolated and expression levels of CD112 were explored by flow cytometry and PCR (Figure 7D). Survival of mice with different combinations of immune checkpoint inhibitors is shown in Figure 7E. Experiments carried out in obese mice with single, double, or triple combinations show that mice treated with triple combinations compared to PDL-1 alone have significantly smaller tumors and better survival. Other immune checkpoint inhibitors, such as anti-PD-1 or anti-CTLA-4, also fail to achieve disease control and are no better than vehicle alone. The addition of iNOS and COX2 inhibitors significantly improves the survival of mice. Mice treated with the triple combination (iNOS, COX2, anti-PD-Ll) increase T cells high in CD226 and low in Tigit. In addition, cancer cells in the tumor microenvironment increase the marker PVRL2 (CD112), the main stimulator of CD226, promoting T cell activation.

[0169] T cell responses to PGE2 and nitric oxide were evaluated. In vitro experiments were used to investigate CD4 or CD8 T cells treated with PGE2 and various PGE2 inhibitors (Figure 8A). CD4 and CD8 T cells treated with nitric oxide were evaluated for expression levels of Tigit (Figure 8B). Figures 8C / D explore the Various combinations of T cells and tumor cells with varied supplementation with PGE2 and nitric oxide were assessed for posttreatment gene expression levels by PCR (Figure 8C ad 8D). PVRL2 expression levels in cancer cells treated with different doses of PGE2 or NO are shown in Figure 8E. In vitro experiments show that stimulation of T cells with either PGE2 or nitric oxide promoted the upregulation of Tigit and downregulation of CD226. This suggests that blocking the production of PGE2 / NO can lead to improved T cell function. T cells treated with PGE2 and PGE2 receptor blocker increased the amount of CD226 expressed on the T cell surface. Cancer cells treated with PGE2 alone decreased the expression of PVRL2 (CD1 12), the costimulatory receptor for CD226.

[0170] Single-cell analysis was also performed on a dataset of human patients who underwent liver resection (Figure 9A). Expression levels of T cell markers by the identity of T cell are shown in Figure 9B. Pathway GO analysis of T cells is shown in Figure 9C. Expression levels of T cells and immunosuppressive or activating markers based on T cell subgroup are shown in Figure 9D. A survival analysis based on the expression levels of EOMES, TBX21, and CD226 is shown in Figure 9E, and correlating genes of interest are shown in Figure 9F. A human gene dataset (GSE15153O) was analyzed based on in vivo findings. T cells with the same expression markers, CD226 high and Tigit low, were found in a subset of patients. These T cells have a similar genomic phenotype compared to T cells found in mice treated with anti-iNOS and anti-COX2. Genes discovered in vitro, including EOMES, TBX21, and CD226, were highly significant for survival in this human cohort.

[0171] Together, these results demonstrate a clear correlation between NOS2 / COX2 expression and immune cell dynamics in HCC and show that co-inhibition of NOS2 and COX2 expression can sensitize HCC cells to immune checkpoint inhibitors.

[0172] Example 3: Correlation ofNOS2 and C0X2 Expression with Tumor Microenvironment and Therapeutic Outcomes in Hepatocellular Carcinoma

[0173] Obese mice with underlying NASH were injected with RIL1-75 HCC cell line into the liver at age 12 weeks. Treatment was started 7 days later. Treatment regimens and dosing were as provided in Table 9. Mice were fed a diet containing GW and CELEBREX®. In addition, similar experiments in mice that grow spontaneous HCC. A STAM model was used in which pups were injected with STZ when they were 2 days old followed by feeding a high fat diet. These mice undergo the same progression as humans: the mice show fatty liver infdtration which progresses to steatosis, to fibrosis, to cirrhosis, to NASH, and finally to HCC. Treatment was started at 20 weeks and was continued until 27 weeks. The only injection the mice received was PDL1, the GW and the CELEBREX® were mixed in the diet. The median survival in days was: Control: 26, CELEBREX®: 30, anti-PDLl: 27.5, CELEB REX®+GW: 27, and CELEBREX®+GW+ anti-PDLl : 47.5.

[0174] Example 4: NOS2 and C0X2 Inhibition and Therapeutic Outcomes

[0175] The results in this Example re-present and expand on at least some of the results provided in other Examples.

[0176] Mice bearing orthotopic liver tumors were established by injecting 600,000 RIL-175 cells directly into the liver. After a 7-day tumor establishment period, animals received various treatment regimens. Treatment regimens and dosing were as provided in Table 9. In the initial experiment (Figure 10A), dual inhibition of inducible nitric oxide synthase (iNOS) using either GW274150 or 1400W in combination with CELEBREX® and anti-PD-Ll significantly improved survival compared to controls or any single-agent treatment.

[0177] Whether combining additional immune checkpoint inhibitors could further enhance therapeutic efficacy was evaluated. As shown in Figure 10B, adding an anti-TIGIT to the regimen (i.e., anti-PD-Ll + GW274150 + CELEBREX® + anti-TIGIT) resulted in superior survival relative to all other treatment arms. Notably, treatment in these studies was halted on day 175, at which time two mice in the quadruple treatment group and one mouse in the anti- PD-Ll + GW274150 + CELEBREX® group were tumor-free at sacrifice. Similar survival benefits were observed when CELEBREX® was replaced by diclofenac, another well- characterized COX-2 inhibitor (Figure 10C). In contrast, anti-PD-1 monotherapy failed to control tumor burden, performing no better than untreated controls (Figure 10D), while similar improvements in survival were also observed with CTLA-4 blockade (Figure 10E).

[0178] To validate these findings across different models, the effects of iNOS / COX-2 inhibition in two additional mouse models of liver cancer were examined. In one model (Figure 11A), 2-day-old C57BL / 6 mice received a single injection of streptozotocin (50 mg / kg) to induce liver cirrhosis and subsequent tumor development by 20 weeks. At 20 weeks, mice were randomized into three groups: untreated controls, anti-PD-Ll alone, or the triple combination of CELEBREX® + GW274150 + anti-PD-Ll. At week 27, control and anti-PD-Ll -treated mice exhibited massive livers with extensive tumor burden, whereas the triple combination completely abrogated macroscopic tumor formation (Figure 1 IB). In a second model (Figure 11C), mice were injected via tail vein with 200,000 RIL-175 cells, and treatment commenced 4 weeks later. Both macroscopic and microscopic evaluations revealed that only the combination of CELEBREX® + GW274150 + anti-PD-Ll achieved complete tumor eradication (Figure 1 ID).

[0179] The impact of dual iNOS / COX-2 inhibition on the tumor immune microenvironment was further explored in the orthotopic model (Figure 12A). As illustrated in Figures 12B- 12C, treatment induced a shift in macrophage polarization, with significantly higher levels of M2 macrophages observed in both non-tumorous liver tissue and within tumors. Moreover, treatment led to a reduction in pro-inflammatory neutrophils and an increase in antigen- presenting dendritic cells within the tumor.

[0180] Complementary in vitro experiments were performed using primary T cells to delineate the molecular effects of inflammatory mediators. Treatment with increasing concentrations of prostaglandin E2 (PGE2; 2 pM and 10 pM) resulted in a dose-dependent downregulation of the activation markers CD226 and T-bet, while TIGIT expression remained unchanged, as determined by qPCR and confirmed by flow cytometry (Figures 13A-13D). In subsequent mechanistic studies, a protein kinase A (PKA) agonist mimicked the effects of PGE2 on CD226 and T-bet expression, whereas co-admini strati on of a PKA inhibitor reversed these effects (Figure 13E). Similarly, exposure to the nitric oxide (NO) donor DETA-NONO decreased CD226 and TBX21 (T-bet) expression while increasing TIGIT levels via activation of the JAK-STAT pathway (Figure 13F). This effect was abrogated by the JAK1 / 2 inhibitor ruxolitinib.

[0181] In a head-to-head comparison of iNOS inhibitors (Figure 14), the triple combination of cindunistat + celecoxib + anti-PD-Ll outperformed the regimen using GW274150 + celecoxib + anti-PD-Ll, although both combinations markedly improved survival relative to untreated controls. Finally, experiments in RAW macrophage cells (Figure 15) demonstrated that LPS-induced nitrite / nitrate secretion was dose-dependently reduced by treatment with various iNOS inhibitors (1400W, GW274150, and cindunistat).

[0182] Collectively, these results indicate that combined inhibition of iNOS and COX-2, particularly when integrated with immune checkpoint blockade, enhances anti-tumor immunity and survival in multiple murine models of liver cancer. These results indicate that combined iNOS / COX-2 inhibition, particularly when paired with immune checkpoint blockade, offers a robust strategy to enhance survival and modulate the tumor immune microenvironment in liver cancer models. OTHER EMBODIMENTS

[0183] It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Claims

WHAT IS CLAIMED IS:

1. A method for treating a mammal having cancer, wherein said method comprises:(a) administering an inhibitor of a nitric oxide synthase (NOS) 2 polypeptide to said mammal;(b) administering an inhibitor of a cyclooxygenase (COX) 2 polypeptide to said mammal; and(c) administering an immune checkpoint inhibitor or a cellular immunotherapy to said mammal.

2. The method of claim 1, wherein said mammal is a human.

3. The method of any one of claims 1-2, wherein said cancer comprises a solid tumor.

4. The method of any one of claims 1-3, wherein said cancer is selected from the group consisting of a liver cancer, a breast cancer, a pancreatic cancer, an ovarian cancer, a lung cancer, a glioblastoma, a melanoma, a colorectal cancer, a brain cancer, an endocrine cancer, a sarcoma, and a testicular cancer.

5. The method of any one of claims 1-4, wherein said inhibitor of said NOS2 polypeptide is selected from the group consisting of 1400W, GW274150, cindunistat (SD- 6010), L-NMMA, and BYK 191023.

6. The method of any one of claims 1-5, wherein said inhibitor of said COX2 polypeptide is selected from the group consisting of celecoxib, etoricoxib, meloxicam, diclofenac, and 2-acetoxybenzoic acid.

7. The method of any one of claims 1-6, wherein said method comprises administering said immune checkpoint inhibitor to said mammal, wherein said immune checkpoint inhibitor is selected from the group consisting of pembrolizumab, nivolumab, cemiplimab,65ipilimumab, tremelimumab, durvalumab, dostarlimab, avelumab, atezolizumab, relatlimab, spartalizumab, camrelizumab, sintilimab, tislelizumab, toripalimab, BMS-8, BMS-37, BMS- 202, BMS-230, BMS-242, BMS-1001, BMS-1166, SB415286, vorinostat, decitabine, entitostat, JQ1, BET151, GSK5O3, panobinostat, ACY-241, azacytidine, DB36, DB71, DB15, CVN, MGCD0103, SNDX-275, IMP32, BMS986016, TSR-022, Sym023, ATIK2a, DZNep, AMP-224, AMP-514, KN035, CK-301, AUNP12, CA-170, BMS-986189, NewEl, and DX11.

8. The method of any one of claims 1-7, wherein said inhibitor of said NOS2 polypeptide is 1400W, wherein said inhibitor of said COX2 polypeptide is celecoxib, and wherein said immune checkpoint inhibitor is selected from the group consisting of atezolizumab, durvalumab, and avelumab.

9. The method of any one of claims 1-8, wherein said method comprises administering said cellular immunotherapy to said mammal, wherein said cellular immunotherapy is selected from the group consisting of administration of autologous T cells, administration of allogenic T cells, a chimeric antigen receptor (CAR) T cell therapy, a tumor-infiltrating lymphocyte (TIL) therapy, a natural killer (NK) cell therapy, administration of a dendritic cell (DC) vaccine, a TCR-engineered T cell therapy, and a macrophage therapy.

10. A method for sensitizing a cancer within a mammal to an immune checkpoint inhibitor, wherein said method comprises:(a) administering an inhibitor of a nitric oxide synthase (NOS) 2 polypeptide to said mammal; and(b) administering an inhibitor of a cyclooxygenase (COX) 2 polypeptide to said mammal.

11. The method of claim 10, wherein said mammal is a human.

12. The method of any one of claims 10-11, wherein said cancer comprises a solid tumor.6613. The method of any one of claims 10-12, wherein said cancer is selected from the group consisting of a liver cancer, a breast cancer, a pancreatic cancer, an ovarian cancer, a lung cancer, a glioblastoma, a melanoma, a colorectal cancer, a brain cancer, an endocrine cancer, a sarcoma, and a testicular cancer.

14. The method of any one of claims 10-13, wherein said inhibitor of said NOS2 polypeptide is selected from the group consisting of 1400W, GW274150, cindunistat (SD- 6010), L-NMMA, and BYK 191023.

15. The method of any one of claims 10-13, wherein said inhibitor of said COX2 polypeptide is selected from the group consisting of celecoxib, etoricoxib, meloxicam, diclofenac, and 2-acetoxybenzoic acid.

16. The method of any one of claims 10-15, wherein said immune checkpoint inhibitor is selected from the group consisting of pembrolizumab, nivolumab, cemiplimab, ipilimumab, tremelimumab, durvalumab, dostarlimab, avelumab, atezolizumab, relatlimab, spartalizumab, camrelizumab, sintilimab, tislelizumab, toripalimab, BMS-8, BMS-37, BMS-202, BMS-230, BMS-242, BMS-1001, BMS-1166, SB415286, vorinostat, decitabine, entitostat, JQ1, BET151, GSK503, panobinostat, ACY-241, azacytidine, DB36, DB71, DB15, CVN, MGCD0103, SNDX-275, IMP32, BMS986016, TSR-022, Sym023, ATIK2a, DZNep, AMP- 224, AMP-514, KN035, CK-301, AUNP12, CA-170, BMS-986189, NewEl, and DX11.

17. The method of any one of claims 10-16, wherein said method further comprises administering said immune checkpoint inhibitor to said mammal.

18. A method for enhancing cellular immunotherapy within a mammal having cancer, wherein said method comprises:(a) administering an inhibitor of a nitric oxide synthase (NOS) 2 polypeptide to said mammal; and67(b) administering an inhibitor of a cyclooxygenase (COX) 2 polypeptide to said mammal.

19. The method of claim 18, wherein said mammal is a human.

20. The method of any one of claims 18-19, wherein said cancer comprises a solid tumor.21 . The method of any one of claims 18-20, wherein said cancer is selected from the group consisting of a liver cancer, a breast cancer, a pancreatic cancer, an ovarian cancer, a lung cancer, a glioblastoma, a melanoma, a colorectal cancer, a brain cancer, an endocrine cancer, a sarcoma, and a testicular cancer.

22. The method of any one of claims 18-21, wherein said inhibitor of said NOS2 polypeptide is selected from the group consisting of 1400W, GW274150, cindunistat (SD- 6010), L-NMMA, and BYK 191023.

23. The method of any one of claims 18-22, wherein said inhibitor of said COX2 polypeptide is selected from the group consisting of celecoxib, etoricoxib, meloxicam, diclofenac, and 2-acetoxybenzoic acid.

24. The method of any one of claims 18-23, wherein said cellular immunotherapy is selected from the group consisting of administration of autologous T cells, administration of allogenic T cells, CAR T cell therapies, TIL therapies, NK cell therapies, DC vaccines, TCR- engineered T cell therapies, and macrophage therapies.

25. The method of any one of claims 18-24, wherein said method further comprises administering said cellular immunotherapy to said mammal.

26. The method of any one of claims 18-25, wherein said method further comprises administering an immune checkpoint inhibitor to said mammal.6827. The method of claim 26, wherein said immune checkpoint inhibitor is selected from the group consisting of pembrolizumab, nivolumab, cemiplimab, ipilimumab, tremelimumab, durvalumab, dostarlimab, avelumab, atezolizumab, relatlimab, spartalizumab, camrelizumab, sintilimab, tislelizumab, toripalimab, BMS-8, BMS-37, BMS-202, BMS-230, BMS-242, BMS-1001, BMS-1166, SB415286, vorinostat, decitabine, entitostat, JQ1, BET151, GSK5O3, panobinostat, ACY-241, azacytidine, DB36, DB71, DB15, CVN, MGCD0103, SNDX-275, IMP32, BMS986016, TSR-022, Sym023, ATIK2a, DZNep, AMP-224, AMP- 514, KN035, CK-301, AUNP12, CA-170, BMS-986189, NewEl, and DX11.

28. A method for treating a mammal having cancer, wherein said method comprises:(a) administering 1400W to said mammal;(b) administering celecoxib to said mammal; and(c) administering an immune checkpoint inhibitor selected from the group consisting of atezolizumab, durvalumab, and avelumab to said mammal.

29. The method of claim 28, wherein said mammal is a human.

30. The method of any one of claims 28-29, wherein said cancer comprises a solid tumor.

31. The method of any one of claims 28-30, wherein said cancer is selected from the group consisting of a liver cancer, a breast cancer, a pancreatic cancer, an ovarian cancer, a lung cancer, a glioblastoma, a melanoma, a colorectal cancer, a brain cancer, an endocrine cancer, a sarcoma, and a testicular cancer.

32. A method for treating a mammal having cancer, wherein said method comprises:(a) administering 1400W to said mammal;(b) administering celecoxib to said mammal; and(c) administering a TIL therapy to said mammal.6933. The method of claim 32, wherein said mammal is a human.

34. The method of any one of claims 32-33, wherein said cancer comprises a solid tumor.

35. The method of any one of claims 32-34, wherein said cancer is selected from the group consisting of a liver cancer, a breast cancer, a pancreatic cancer, an ovarian cancer, a lung cancer, a glioblastoma, a melanoma, a colorectal cancer, a brain cancer, an endocrine cancer, a sarcoma, and a testicular cancer.70

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