Combination of tyrosine kinase inhibitors and pro-inflammatory agents for treatment of cancer

Abstract

The present application provides methods of treating cancer in a subject comprising administering to the subject a tyrosine kinase inhibitor and a pro-inflammatory agent, such as a TLR agonist, a STING activator, radiotherapy, or an immune checkpoint inhibitor. In some cases, the methods include administering a tyrosine kinase inhibitor when the individual is in an inflammatory response.

Description

[0001] Cross Reference to Related Applications

[0002] This application claims the benefit of and priority to U.S. Provisional Application No. 63 / 382,003, filed November 2, 2022, U.S. Provisional Application No. 63 / 491,000, filed March 17, 2023, and U.S. Provisional Application No. 63 / 581,197, filed September 7, 2023, the contents of each of which are incorporated by reference herein in their entirety. TECHNICAL FIELD

[0003] The present invention relates to compositions and methods for treating cancer, the methods comprising administration of a tyrosine kinase inhibitor and optionally a proinflammatory agent. BACKGROUND

[0005] Intratumoral myeloid leukocytes, including macrophages (i.e., tumor-associated macrophages or TAMs) and myeloid-derived suppressor cells (MDSCs), play a key role in controlling the tumor microenvironment (TME) immunosuppression that supports tumor growth and also confers resistance to immunotherapy in cancers such as solid tumors. One important mechanism by which myeloid leukocytes assume an immunosuppressive phenotype or enhance their immunosuppressive capacity upon tumor therapy is through their cell surface inhibitory receptors (iRs) that, when activated, are also driven by their extracellular ligand binding that triggers multiple pathways of negative regulation via their cytoplasmic domains based on immunoreceptor tyrosine-based inhibitory motifs (ITIMs) that activate the central signaling regulator SHP-1 to dephosphorylate, thereby inactivating many signal transduction molecules. This reduces the therapeutic-induced anticancer proinflammatory response Figure 1 ). In solid tumors, the expression of essential cell surface iRs (such as SIRPa, Siglecs family, LilRB and PirB family, LAIR1, lectin receptor family, SLAM receptor family, etc.) that are shown to increase with tumor progression in the TME are regulated via activation of SHP-1, which then mediates downstream suppression.

[0006] In view of these inhibitory mechanisms elucidated over the past few years, therapeutic development pipelines aimed at blocking iRs (e.g., anti-LilRB1 / 2 and anti-SIRPa) and their ligands (e.g., anti-CD47) are being advanced (3-5). However, these efforts that individually target each iR or its ligand, rather than simultaneously targeting all inhibitory pathways, have achieved weak to partial efficacy in controlling solid tumors.

[0007] The disclosures of all publications, patents, patent applications, and published patent applications referred to in this document are hereby incorporated by reference in their entirety. SUMMARY

[0008] The present application provides, in one aspect, a method of treating a cancer in an individual comprising administering to the individual a) a tyrosine kinase inhibitor, and b) a proinflammatory agent, optionally wherein the method comprises administering the tyrosine kinase inhibitor to the individual intermittently. In some embodiments, the method comprises administering the tyrosine kinase inhibitor systemically or locally (e.g., intratumorally). In some embodiments, the proinflammatory agent comprises an agent selected from the group consisting of a TLR agonist, a STING activator, a radiation therapy, a PAMP / DAMP molecule, a checkpoint inhibitor, a proinflammatory cytokine, a proinflammatory cell, a cell, a cancer vaccine, a chemotherapeutic agent, a bacterial component, a cancer vaccine, an oncolytic virus, a sound therapy, a magnetotherapy, an electrotherapy, and an electrostatic therapy.

[0009] The present application provides, in another aspect, a method of treating a cancer in an individual comprising administering to the individual a) a tyrosine kinase inhibitor, and b) a proinflammatory agent, wherein the method comprises administering the tyrosine kinase inhibitor systemically. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual intermittently. In some embodiments, the proinflammatory agent comprises an agent selected from the group consisting of a TLR agonist, a STING activator, a radiation therapy, a PAMP / DAMP activator, a checkpoint inhibitor, a proinflammatory cytokine, a chemotherapeutic agent, a bacterial component, a cancer vaccine, an oncolytic virus, a sound therapy, a magnetotherapy, an electrotherapy, and an electrostatic therapy.

[0010] The present application provides, in another aspect, a method of treating a cancer in an individual comprising administering to the individual a) a tyrosine kinase inhibitor, and b) a proinflammatory agent, and wherein the proinflammatory agent comprises an agent selected from the group consisting of a TLR agonist, a STING activator, a PAMP / DAMP activator, a chemotherapeutic, a proinflammatory cytokine, a cancer vaccine, a bacterial component, a sound therapy, a magnetotherapy, an electrotherapy, and an electrostatic therapy. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual intermittently. In some embodiments, the method comprises administering the tyrosine kinase inhibitor systemically.

[0011] The present application provides, in another aspect, a method of treating a cancer in an individual comprising administering to the individual a tyrosine kinase inhibitor, wherein the individual is in an inflammatory response or has an ongoing infection. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual intermittently. In some embodiments, the method comprises administering the tyrosine kinase inhibitor systemically. In some embodiments, the method further comprises an immune cell.

[0012] In some embodiments according to any of the methods described above, the method comprises administering the tyrosine kinase inhibitor to the individual at an interval of no more than once every three days at least twice.

[0013] In some embodiments according to any of the above methods, the method comprises administering the tyrosine kinase inhibitor to the individual for at least two cycles, wherein each cycle is from about three days to about twenty days.

[0014] In some embodiments according to any of the above methods, the tyrosine kinase inhibitor has a half-life of no more than about 5 days, optionally wherein the tyrosine kinase inhibitor has a half-life of no more than about 3 days.

[0015] In some embodiments according to any of the above methods, the tyrosine kinase inhibitor is effective to inhibit tyrosine kinase activity by more than 50% for no more than about 5 days, optionally wherein the tyrosine kinase inhibitor is effective to inhibit tyrosine kinase activity by more than 50% for no more than about 3 days.

[0016] In some embodiments according to any of the above methods, the tyrosine kinase inhibitor is selected from the group consisting of small molecules, nucleic acids (e.g., siRNA, shRNA, antisense RNA, microRNA), nucleic acid editing systems (e.g., CRISPR systems), and protein agents (e.g., antibody agents that target tyrosine kinases or activated tyrosine kinases). In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling. In some embodiments, the tyrosine kinase inhibitor inhibits Src family kinases (SFKs). In some embodiments, the tyrosine kinase inhibitor is an Hck inhibitor. In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406.

[0017] In some embodiments according to any of the above methods, the tyrosine kinase inhibitor is administered at least three times. In some embodiments according to any of the above methods, the method comprises administering the tyrosine kinase inhibitor systemically and locally, optionally wherein the method comprises administering the tyrosine kinase inhibitor intratumorally.

[0018] In some embodiments according to any of the above methods, the systemic administration of the tyrosine kinase comprises oral administration, intravenous administration, subcutaneous administration, and / or intraperitoneal administration.

[0019] In some embodiments according to any of the above methods, the pro-inflammatory agent and the tyrosine kinase inhibitor are administered within about 24 hours of each other (e.g., within about 16 hours, 8 hours, 4 hours, 2 hours, 1 hour, or 0.5 hours of each other).

[0020] In some embodiments according to any of the above methods, the method comprises administering the pro-inflammatory agent intratumorally.

[0021] In some embodiments according to any of the above methods, the method comprises administering a pro-inflammatory agent to a site different from the site of the cancer to be treated.

[0022] In some embodiments according to any of the above methods, the pro-inflammatory agent comprises a TLR agonist. In some embodiments, the TLR agonist activates a TLR on a macrophage. In some embodiments, the TLR comprises TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, and / or TLR9. In some embodiments, the TLR agonist comprises CpG, poly I:C, and / or R848, flagellin (TLR5), zymosan (TLR2 / 4), radiation therapy generated DAMPs such as HMGB1 (TLR2 / 4), DNA and RNA molecules (TLR3 / 7 / 8 / 9), etc. In some embodiments, the TLR agonist comprises CpG, poly I:C, and R848, e.g., at a ratio of 1 : 1 : 1.

[0023] In some embodiments according to any of the above methods, the pro-inflammatory agent comprises a bacterial component, optionally the bacterial component comprises lipopolysaccharide (LPS).

[0024] In some embodiments according to any of the above methods, the pro-inflammatory agent comprises a STING activator. In some embodiments, the STING activator comprises 2'3'-cGAMP.

[0025] In some embodiments according to any of the above methods, the pro-inflammatory agent comprises a chemotherapeutic agent. In some embodiments, the chemotherapeutic comprises azathioprine (AZA).

[0026] In some embodiments according to any of the above methods, the pro-inflammatory agent comprises a pro-inflammatory cytokine. In some embodiments, the pro-inflammatory cytokine comprises IL-1 family cytokines (e.g., IL-1b, IL-18), IL-6, IL-17, TNF family cytokines (e.g., TNFa), and combinations thereof with type I and type II interferons (IFNa, IFNb, and IFNy).

[0027] In some embodiments according to any of the above methods as applied, the pro-inflammatory agent comprises radiation therapy. In some embodiments, the radiation therapy comprises irradiation at the site of the cancer to be treated. In some embodiments, the radiation therapy comprises irradiation at a site different from the site of the cancer to be treated. In some embodiments, the dose of radiation therapy is non-ablative, insufficient to eliminate the tumor (kill all tumor cells).

[0028] In some embodiments according to any of the above methods, the proinflammatory agent comprises a checkpoint inhibitor. In some embodiments, the checkpoint inhibitor comprises an anti-PD-Ll antibody, an anti-PD-1 antibody, or an anti-CLTA4 antibody.

[0029] In some embodiments according to any of the above methods, the proinflammatory agent is administered intermittently.

[0030] In some embodiments according to any of the above methods, the proinflammatory agent and the tyrosine kinase inhibitor are administered simultaneously or concurrently.

[0031] In some embodiments according to any of the above methods, the proinflammatory agent comprises an immune cell. In some embodiments, the immune cell is derived from the same individual. In some embodiments, the immune cell comprises or is a macrophage, optionally wherein the macrophage has a proinflammatory (Ml) phenotype. In some embodiments, the immune cell is derived from a monocyte. In some embodiments, the immune cell expresses high levels of MHC-I, MHC-II, CD80, and / or CD86. In some embodiments, the immune cell expresses one or more proinflammatory cytokines, optionally wherein the one or more proinflammatory cytokines comprise TNFa and / or IL-12. In some embodiments, the immune cell does not express significant levels of TGF and / or IL-10. In some embodiments, the immune cell comprises a T cell. In some embodiments, the immune cell is engineered to express a chimeric antigen receptor, optionally wherein the chimeric antigen receptor specifically binds to a tumor antigen. In some embodiments, the macrophage is engineered to be tyrosine kinase expression and / or activation deficient. In some embodiments, the tyrosine kinase inhibitor and the immune cell are administered within 24 hours of each other, optionally wherein the tyrosine kinase inhibitor and the immune cell are administered within 4 hours of each other. In some embodiments, the immune cell is administered simultaneously or concurrently with the tyrosine kinase inhibitor.

[0032] In some embodiments according to any of the above methods, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof). In some embodiments, the SHP-1 is administered simultaneously with the tyrosine kinase inhibitor. In some embodiments, the SHP-1 is administered sequentially (e.g., before or after the tyrosine kinase inhibitor). In some embodiments, the SHP-1 administration follows the same dosing regimen as the tyrosine kinase inhibitor.

[0033] In some embodiments according to any of the above methods, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm, including but not limited to anti-TNFa antibodies and anti-IL6 antibodies. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered prior to (e.g., within any of about one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day or less, prior to) the tyrosine kinase inhibitor. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered sequentially (e.g., prior to or after the tyrosine kinase inhibitor). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered immediately after (e.g., within any of about 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours, after) the tyrosine kinase inhibitor. In some embodiments, the administration of the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm follows the same dosing regimen as the tyrosine kinase inhibitor.

[0034] In some embodiments according to any of the above methods, the cancer is a solid tumor.

[0035] In some embodiments according to any of the above methods, the cancer is a hematological cancer.

[0036] In some embodiments according to any of the above methods, the cancer is an end-stage cancer.

[0037] In some embodiments according to any of the above methods, the cancer is resistant or refractory to radiation therapy, a chemotherapeutic agent, and / or a checkpoint inhibitor.

[0038] In some embodiments according to any of the above methods, the individual is a human.

[0039] The present application provides, in another aspect, a composition comprising a tyrosine kinase inhibitor and a proinflammatory agent, optionally wherein the proinflammatory agent comprises an agent selected from the group consisting of an immune cell, a TLR agonist, a STING activator, an agent for use in radiation therapy, a PAMP / DAMP activator, a checkpoint inhibitor, a proinflammatory cytokine, a chemotherapeutic agent, a bacterial component, a cancer vaccine, an oncolytic virus, and an agent for use in sound therapy, magnetotherapy, electrotherapy, or electrostatic therapy. BRIEF DESCRIPTION OF DRAWINGS

[0040] Figure 1 SHP-1 is depicted as a “master” signaling mediator downstream of multiple inhibitory receptors on myeloid leukocytes in the tumor microenvironment (TME). Diminished activity of SHP-1 attenuates RT and immunotherapy-induced proinflammatory pathways and anticancer efficacy, and maintains a myeloid leukocyte immunosuppressive phenotype. Methods of tyrosine kinase inhibition as an anticancer strategy are marked with a circle. Partially listed are companies and methods aimed at depleting or blocking individual cell surface inhibitory receptors, including SIRPa (SIRPa nt Immunotherapeutics, anti-CD47 Gilead) and anti-SIRPa (Biosion) methods), Siglecs (NextCure), LilRB (Next-IO), SLAMF (BMS), and others.

[0041] Figures 2A-2E Sample assays for in vitro studies of RK-20449 and dasatinib are shown. Figure 2A In vitro assay systems are shown. Figure 2B RK20449 and dasatinib dose-dependently reduce macrophage SHP-1 activity induced by aT LR and cancer cell ligation are shown. Figure 2C RK20449 and dasatinib deplete aT LR and cancer cell ligation-induced iR phosphorylation and binding to SHP-1 are shown. Figure 2D and Figure 2E RK-20449 or dasatinib treatment enables macrophages to overcome the suppression exerted by tumor cells to release proinflammatory phenotype expression induced by aT LR, demonstrating a significant increase in production of proinflammatory cytokines TNFa, IL-6, and CXCL1 (panel D) and expression of cell surface antigen-presenting mechanisms (panel E) are shown. Figure 2E In parallel experiments, the SHI-1 inhibitor TPI-1 was used, which also exhibited similar effects.

[0042] Figures 3A-3G Effects of RK-20449 or dasatinib in combination with TLR agonists on solid tumors are shown. Figure 3A and Figure 3F The upper panel of shows the experimental design. Figure 3BEfficacy of tyrosine kinase inhibitor (TKi) treatment alone or in combination with TLR agonists on MC38 colorectal carcinoma is shown. Figure 3C and Figure 3F Efficacy of TKi treatment alone or in combination with TLR agonists on KPC pancreatic ductal adenocarcinoma is shown. Figure 3D Dose-dependent effects of TKi on LLC lung carcinoma are shown. Figure 3E Efficacy of dasatinib in combination with various TLR agonists for treatment of MC38 colorectal carcinoma is shown. Figure 3G Inhibition of TK reduces tumor angiogenesis. TKi such as dasatinib inhibit angiogenesis in tumors. This effect is achieved by inhibiting VEGFR, a receptor tyrosine kinase.

[0043] Figures 4A-4B Dasatinib and aTLR in combination induce anti-tumor T cell immunity is shown. Figure 4A Flow cytometry analysis of the frequency of immune cells (CD45+) within the total cell population of tumor dissociates and the frequency of individual immune cell types labeled by specific antibodies is shown. Figure 4B Summary of the dynamics of each immune population is shown. Dasatinib, once combined with aTLR, exhibits a dose-dependent effect of increasing CD8+ T cells and NK cells and decreasing macrophages and MDSCs within the TME.

[0044] Figures 5A-5C Efficacy of RK-20449 or dasatinib in combination with a Sting activator (VISTA), Figure 5A tumor lesion RT ( Figure 5B ), or pro-inflammatory cytokines ( Figure 5C ) for treatment of MC38 colorectal carcinoma is shown.

[0045] Figures 6A-6C PD-1 / PD-L1 immune checkpoint blockade enhances the efficacy of dasatinib and aTLR combination therapy is shown. Figure 6A Experimental protocol is shown. Figure 6B Luminescence images showing tumor location and size are shown. Figure 6C Tumor volume change after treatment is shown.

[0046] Figures 7A-7C In vitro testing of UM-164, R406, piceatannol, baricitinib, and ibrutinib for reducing the TK-iRS-SHP-1 axis in TAMs induced by aTLR and cancer cell ligation is shown. Various TKi are tested for their ability to reduce SHP-1 activity in macrophages induced by aTLR and cancer cell ligation Figure 7A ), their ability to improve antigen presentation mechanisms Figure 7B , and their ability to induce pro-inflammatory cytokinesFigure 7C ).

[0047] Figures 8A-8C In vivo testing of the anti-tumor efficacy of R406, UM-164, whitbyol, and SHP inhibitor 3Ac in combination with aTLR is shown. Figure 8A R406, but not the other inhibitors, is shown to be effective in combination with aTLR to inhibit LLC tumors. Figure 8B TME analysis is shown demonstrating that R406 in combination with aTLR induces expansion of CD8+ T cells within the tumor. Figure 8C R406 in combination with aTLR is shown to induce antigen presentation by macrophages within the tumor.

[0048] Figures 9A-9C Anti-cancer effects of TK inhibitors ponatinib, bosutinib, saracatinib, and KX2-391 are shown. Four TK inhibitors including ponatinib, bosutinib, saracatinib, and KX2-391 were tested for their ability to reduce macrophage SHP-1 activity induced by aTLR and cancer cell ligation ( Figure 9A ), and to increase expression of antigen presentation molecules otherwise suppressed by cancer cell ligation ( Figure 9B ). In vivo anti-tumor efficacy of aTLR combinations is shown in ( Figure 9C ).

[0049] Figures 10A-10E Synergy of SHP-1 inhibitor TPI-1 and TK inhibitor dasatinib is shown. Figure 10A An experimental design to test treatments against KPC is shown. Figure 10B KPC tumor volume changes after treatment with aTLR plus TPI-1 or aTLR plus TPI-1 and dasatinib are shown compared to no treatment (NT) and tumors showing continued progression. Figure 10C Results of TME analysis are shown. aTLR plus TPI-1 and dasatinib further enhance T cell immunity while reducing PMN infiltration compared to aTLR plus TPI-1, resulting in increased CD8 (Tc) and CD4 (Th) T cells, and modestly increased NK cells, but reduced PMN in the TME after treatment. Figure 10D An experimental design to test treatments against MC38 is shown. Figure 10E MC38 tumor volume changes after treatment with aTLR plus TPI-1, aTLR plus dasatinib, or aTLR plus TPI-1 and dasatinib are shown compared to no treatment (NT) and tumors showing continued progression.

[0050] Figures 11A-11G Anti-TNFa mAb is shown to inhibit systemic inflammation and reduce adverse toxicities. Figure 11AThe experimental design is shown. Patients with established MC38 colorectal cancer (200-400 mm) were treated with αTLR, TPI-1, and dasatinib (subcutaneously) without additional treatment, or with anti-TNFα mAb or anti-IL-6 mAb (150 μg, intraperitoneal). 3 Mice were used. Treatment was repeated once (day 1 and day 2). Tumor volume changes were recorded, and immune infiltration of the tumor TME was analyzed on day 6 post-treatment. Figure 11B The changes in tumor volume after various treatments are shown. Figure 11C and Figure 11D The results of the TME analysis are shown. Treatment with anti-TNFα mAb or anti-IL-6 mAb did not affect the increase in CD8 T cells (Tc) and NK cells induced by αTLR / TPI-1 / dasatinib treatment, nor the decrease in macrophages and MDSCs in the TME. Figure 11E The study showed that treating mice with anti-TNFα mAb instead of anti-IL-6 mAb significantly reduced the induction of inflammatory cytokines (TNFα, IL-6, IL-1β, IL-10, IFNα, and IFNγ) associated with combination therapy with αTLR / TPI-1 / dasatinib. Figure 11F The study showed that anti-TNFα treatment also significantly reduced circulating monocytes and PMN chemokines CCL2, CCL5, and CXCL1, without reducing CXCL10, which is essential for T cell transport. Figure 11G Anti-TNFα treatment was shown to protect mice from developing splenomegaly and intestinal inflammation, which are typically associated with αTLR / TPI-1 / dasatinib treatment.

[0051] Figure 12 This study demonstrates the proprietary binding of pro-inflammatory stimuli (TLR agonists, pro-inflammatory cytokines IL-1β, IL-6, IL-12, IL-17, IL-18, TNFα, IFNγ, etc., and cancer therapy) to SIRPαITIM phosphorylation and SHP-1 (not SHP-2). Detailed Implementation

[0052] The present application provides, in one aspect, a method of treating cancer in an individual comprising administering to the individual a tyrosine kinase inhibitor, wherein the individual a) has received, is receiving, or will receive a proinflammatory agent, or b) is in an inflammatory response or has an ongoing infection. The present application provides, in another aspect, a method of treating cancer in an individual comprising administering to the individual a monocyte or macrophage deficient in tyrosine kinase expression or activation, and wherein the individual a) has received, is receiving, or will receive a proinflammatory agent, or b) is in an inflammatory response or has an ongoing infection. In some embodiments, the tyrosine kinase inhibitor is administered systemically. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual at least twice at intervals of no more than once every three days. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual for at least two cycles, wherein the tyrosine kinase inhibitor is administered at least once in each cycle, and wherein each cycle has about three days to about twenty days. In some embodiments, the proinflammatory agent comprises an agent selected from the group consisting of a TLR agonist, a STING activator, a radiation therapy, a PAMP / DAMP activator, a checkpoint inhibitor, a proinflammatory cytokine, a chemotherapeutic agent, a bacterial component, a cancer vaccine, an oncolytic virus, a radiation therapy, a sound therapy, a magnet therapy, an electric therapy, and an electrostatic therapy. Further combination therapy methods are provided.

[0053] The present application is based, at least in part, on the surprising discovery that the combination of a tyrosine kinase inhibitor that potentially inhibits the activation of the “master” suppressor SHP-1 with a proinflammatory therapy releases proinflammatory signaling in the tumor environment, particularly acting on tumor infiltrating macrophages, resulting in a dramatic reprogramming of the TME and enhanced activation of innate and adaptive immune cells to promote anti-cancer immunity. Specifically, it was discovered that iR-SHP-1 mediated suppressive regulation within the tumor is particularly strong under tumor therapy, as these therapies often induce ITIM hyperphosphorylation, thereby stimulating ‘over-activation’ of SHP-1, a feedback loop that protects tumors from therapy damage and inflammatory attack, and also elicits wound healing responses to promote tumor progression. This discovery highlights the potential of inhibiting upstream tyrosine kinases that can deplete ITIM phosphorylation and SHP-1 activation as a combination in tumor immunotherapy to achieve efficacy. This approach depletes ITIM phosphorylation and SHP-1 activation by inactivating TK activity, thereby enabling cancer therapy.

[0054] It has been demonstrated that the combination of a tyrosine kinase inhibitor with a proinflammatory agent, such as a TLR agonist and / or a checkpoint inhibitor, achieves a remarkable effect of transforming an immunosuppressive TME into an inflammatory TME, energizing various types of immune cells, such as macrophages, T cells, and B cells, and completely depleting tumors. See, e.g., Figures 3B-3E , Figures 4A-4B and Figures 5A-5C .

[0055] It was also discovered that the tyrosine kinase inhibitors and TLR agonists further enhance their therapeutic efficacy when combined with immune checkpoint inhibitors (e.g., anti-PD-Ll inhibitors) to accelerate tumor regression. See Figures 6A-6C .

[0056] In addition, the tyrosine kinase inhibitors and TLR agonists show synergistic effects in various tumor models when combined with SHP-1 inhibitors. See Figures 10A-10E The administration of agents that reduce systemic inflammation (e.g., anti-TNFa mAbs) further suppress systemic inflammation and reduce adverse toxicities. See Figures 11A-11G .

[0057] Accordingly, the present application provides new methods that can effectively reprogram the immunosuppression caused by the tumor environment and allow both innate and adaptive immunity against cancer, resulting in significant anti-tumor efficacy.

[0058] I. DEFINITIONS

[0059] In general, the terms used in the claims and the specification should be construed to have their ordinary meaning as understood by those with ordinary skill in the art. To the extent that any definitions provided herein conflict with the ordinary meaning of a term, the definitions provided herein should be controlling.

[0060] The terms "individual," "subject," or "patient" are used synonymously herein to describe a mammal, including a human. An individual includes, but is not limited to, a human, a bovine, a horse, a cat, a dog, a rodent, or a primate. In some embodiments, the individual is a human. In some embodiments, the individual has a disease, such as cancer. In some embodiments, the individual is in need of treatment.

[0061] "Reference," as used herein, refers to any sample, standard, or level used for comparison purposes. The reference can be obtained from a healthy and / or non-diseased sample. In some examples, the reference can be obtained from an untreated sample. In some examples, the reference is obtained from a non-diseased or untreated sample of the individual. In some examples, the reference is obtained from the individual or one or more healthy individuals of the individual.

[0062] The term "intermittent" or "intermittently," as used herein, in the context of dosing, refers to non-continuous dosing. In some cases, "intermittent" dosing refers to dosing where a) the tyrosine kinase inhibitor is administered for less than 12 consecutive days (e.g., less than 11 days, 10 days, 9 days, 8 days, 7 days, 6 days, 5 days, 4 days, and 3 days), and b) the tyrosine kinase inhibitor is administered at least twice, and the two administrations are separated by at least one day (i.e., on day 1 and day 3).

[0063] As used herein, the term "cycle" in the context of dosing refers to a period of time during which there is at least one administration of a tyrosine kinase inhibitor. Day 1 of a cycle is defined as the day on which the first administration of a tyrosine kinase inhibitor occurs during that period of time. When there are several consecutive days of administration of a tyrosine kinase inhibitor, day 1 of a cycle is defined as the day on which the first administration in the several consecutive days of administration occurs. The last day of a cycle is defined as the day preceding the occurrence of the next non-consecutive administration of a tyrosine kinase inhibitor. See, e.g., FIGS. 14A and 14B. These cycles need not have the same length of time. For example, a first cycle can have five days and a second cycle has seven days. Each cycle can have a different number of administrations of a tyrosine kinase inhibitor. For example, a first cycle that can have five days can have one administration of a tyrosine kinase inhibitor and a second cycle that can have seven days can have two administrations of a tyrosine kinase inhibitor. Figure 12 A and FIG. 14A. These cycles need not have the same length of time. For example, a first cycle can have five days and a second cycle has seven days. Each cycle can have a different number of administrations of a tyrosine kinase inhibitor. For example, a first cycle that can have five days can have one administration of a tyrosine kinase inhibitor and a second cycle that can have seven days can have two administrations of a tyrosine kinase inhibitor.

[0064] As used herein, the term "immunogenicity" is the ability to elicit an immune response, e.g., via T cells, B cells, or both.

[0065] As used herein, "treatment" or "treating" is an approach for obtaining beneficial or desired results, including clinical results. For the purposes of this application, beneficial or desired clinical results include, but are not limited to, one or more of the following: reducing one or more symptoms resulting from the disease, diminishing the extent of the disease, stabilizing the disease (e.g., preventing or delaying the worsening of the disease), preventing or delaying the spread (e.g., metastasis) of the disease, preventing or delaying the onset of the disease or recurrence of the disease, delaying or slowing the progression of the disease, ameliorating the disease state, providing a partial or complete resolution of a disease, reducing the dose of one or more other medications required to treat the disease, delaying the progression of the disease, increasing the quality of life, and / or increasing the survival period. "Treatment" also encompasses reduction of pathological consequences of a cancer. The methods of the application encompass any one or more of these treatment aspects.

[0066] As used herein, "delaying" development of a cancer means to defer, hinder, slow, retard, stabilize, and / or postpone development of the disease. This delay can be of varying lengths, specific to the history of the disease and / or individual being treated. As will be apparent, a sufficient or significant delay can in effect encompass prevention, in that the individual does not develop the disease. A method that "delays" development of a cancer is one that reduces the likelihood of development of the disease within a given timeframe and / or reduces the extent of the disease within a given timeframe as compared to not using the method. Such comparisons are typically based on clinical studies using statistically significant numbers of individuals. Cancer development can be detected using standard methods, including but not limited to computed axial tomography (CAT scan), magnetic resonance imaging (MRI), abdominal ultrasound, coagulation testing, arteriography, or biopsy. Development can also refer to progression of a cancer that can not initially be detectable and includes onset, recurrence, and attack.

[0067] As used herein, the term "simultaneous administration" means that the first treatment and the second treatment in a combination therapy are administered with a time interval of no more than about 15 minutes, such as no more than about any of 10 minutes, 5 minutes, or 1 minute. When the first treatment and the second treatment are administered simultaneously, the first treatment and the second treatment can be contained in the same composition (e.g., a composition containing both the first treatment and the second treatment) or in separate compositions (e.g., the first treatment is contained in one composition and the second treatment is contained in another composition).

[0068] As used herein, the term "sequential administration" means that the first treatment and the second treatment in a combination therapy are administered with a time interval of more than about 15 minutes, such as more than about any of 20 minutes, 30 minutes, 40 minutes, 50 minutes, 60 minutes, or more. Either the first treatment or the second treatment can be implemented first. The first treatment and the second treatment are contained in separate compositions, which can be contained in the same or different packaging or kits.

[0069] As used herein, the term "concurrent administration" means that the administration of the first treatment and the administration of the second treatment in a combination therapy overlap each other.

[0070] As used herein, by "pharmaceutically acceptable" or "pharmacologically compatible" is meant a material that is not biologically or otherwise undesirable, e.g., the material can be incorporated into a pharmaceutical composition administered to an individual without any appreciable deleterious effects and without interacting in a deleterious manner with any of the other components of the composition in which it is contained. Pharmaceutically acceptable carriers or excipients preferably meet the required standards of toxicology and manufacturing testing, and / or are included in the U.S. Food and Drug Administration's guidelines for inactive ingredients.

[0071] It should be understood that the embodiments of this application described herein include “consisting of embodiments” and / or “substantially consisting of embodiments”.

[0072] In this document, mentioning "about" a value or parameter includes (and describes) variations relating to that value or parameter itself. For example, a description of "about X" includes a description of "X".

[0073] As used in this article, mentioning "not" a value or parameter generally means and describes something that is "different" from a value or parameter. For example, "the method is not used to treat type X cancer" means that the method is used to treat a type of cancer different from X.

[0074] The term “about XY” as used in this article has the same meaning as “about X to about Y”.

[0075] It should be noted that, unless the context clearly specifies otherwise, the singular forms “a,” “an,” and “the” used in the specification and appended claims include a plurality of indicators.

[0076] Any term not directly defined herein should be understood to have the meaning generally associated with it as understood within the field of this invention. Certain terms are discussed herein to provide additional guidance to practitioners in describing compositions, apparatuses, methods, etc., and how they are made or used in relation to aspects of the invention. It should be understood that the same thing can be expressed in more than one way. Therefore, alternative language and synonyms may be used for any one or more of the terms discussed herein. Whether a term is explicitly stated or discussed herein is not significant. Several synonyms or alternative methods, materials, etc., are provided. Unless explicitly stated, the use of one or more synonyms or equivalents does not preclude the use of other synonyms or equivalents. The use of examples, including instances of terms, is for illustrative purposes only and does not limit the scope and meaning of aspects of the invention.

[0077] II. Treatment methods

[0078] This application provides, in one aspect, a method for treating cancer by administering a tyrosine kinase inhibitor. In some embodiments, the individual being treated has received, is receiving, or will receive a pro-inflammatory agent, such as any of those described herein. In some embodiments, the individual is in an inflammatory response or has an ongoing infection.

[0079] In some embodiments, the method includes administering both a tyrosine kinase inhibitor and a pro-inflammatory agent to an individual. In some embodiments, the tyrosine kinase inhibitor is administered intermittently. In some embodiments, the method includes systemic administration of the tyrosine kinase inhibitor.

[0080] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor, wherein the individual a) has received, is receiving, or will receive a pro-inflammatory agent (e.g., a TLR agonist, e.g., radiation therapy), or b) is in an inflammatory response or has an ongoing infection, and optionally wherein the tyrosine kinase inhibitor is administered systemically (e.g., intravenously or subcutaneously). In some embodiments, the tyrosine kinase inhibitor is administered intermittently. In some embodiments, the tyrosine kinase inhibitor is administered at intervals of no more than once every two days. In some embodiments, the tyrosine kinase inhibitor is administered no less than two and no more than 5 times within a consecutive ten days (e.g., twice within ten days, three times within ten days, four times within ten days, or five times within ten days). In some embodiments, the tyrosine kinase inhibitor is administered concurrently with the pro-inflammatory agent. In some embodiments, the tyrosine kinase inhibitor is administered concurrently with the pro-inflammatory agent. In some embodiments, the tyrosine kinase inhibitor and the pro-inflammatory agent are administered sequentially within 2 weeks (e.g., within 10 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or the same day). In some embodiments, the tyrosine kinase inhibitor has a half-life of no more than about 10 days (e.g., no more than about 7 days, 5 days, 4 days, or 3 days). In some embodiments, the tyrosine kinase inhibitor is effective to inhibit more than 50% of tyrosine kinase activity for no more than about 7 days (e.g., about 5 days, 4 days, or 3 days). In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of a small molecule, a nucleic acid (e.g., siRNA, shRNA, antisense RNA, microRNA), a nucleic acid editing system (e.g., a CRISPR system), and a protein agent (e.g., an antibody agent that targets a tyrosine kinase or an activated tyrosine kinase). In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling. In some embodiments, the tyrosine kinase inhibitor inhibits a Src family kinase (SFK). In some embodiments, the tyrosine kinase inhibitor is an Hck inhibitor. In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406. In some embodiments, the method further comprises administering the pro-inflammatory agent locally (e.g., intratumorally) to the individual. In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof). In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody).In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered prior to (e.g., within about any of one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to the tyrosine kinase inhibitor). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered concurrently with the tyrosine kinase inhibitor. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered concurrently with the tyrosine kinase inhibitor. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered sequentially (e.g., prior to or after the tyrosine kinase inhibitor). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered immediately after (e.g., within about any of 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours, after the tyrosine kinase inhibitor). In some embodiments, the TLR agonist is selected from the group consisting of LTA, CpG, Poly I;C, LPS, MPLA, flagellin, R848, resiquimod, bromopindine, motolimod, and loxoribine.

[0081] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and a proinflammatory agent (e.g., a TLR agonist, e.g., radiation therapy), and wherein the method optionally comprises oral, intravenous, or subcutaneous administration of the tyrosine kinase inhibitor, optionally wherein the tyrosine kinase inhibitor is administered intermittently. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual at an interval of no more than once every three days at least twice. In some embodiments, the tyrosine kinase inhibitor is administered twice (e.g., two on days) every seven to twenty days. In some embodiments, the tyrosine kinase inhibitor is administered three times (e.g., three on days) every ten to twenty days. In some embodiments, the tyrosine kinase inhibitor is administered at an interval of no more than once every two days. In some embodiments, the tyrosine kinase inhibitor is administered no less than two and no more than 5 times within a consecutive ten days (e.g., twice within ten days, three times within ten days, four times within ten days, or five times within ten days). In some embodiments, the tyrosine kinase inhibitor is administered concurrently with the proinflammatory agent. In some embodiments, the tyrosine kinase inhibitor is administered concurrently with the proinflammatory agent. In some embodiments, the tyrosine kinase inhibitor and the proinflammatory agent are administered sequentially within 2 weeks (e.g., within 10 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or the same day). In some embodiments, the tyrosine kinase inhibitor has a half-life of no more than about 10 days (e.g., no more than about 7 days, 5 days, 4 days, or 3 days). In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of a small molecule, a nucleic acid (e.g., siRNA, shRNA, antisense RNA, microRNA), a nucleic acid editing system (e.g., a CRISPR system), and a protein agent (e.g., an antibody agent that targets a tyrosine kinase or an activated tyrosine kinase). In some embodiments, the tyrosine kinase inhibitor is a Src inhibitor. In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling. In some embodiments, the tyrosine kinase inhibitor inhibits a Src family kinase (SFK). In some embodiments, the tyrosine kinase inhibitor is a Hck inhibitor. In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406. In some embodiments, the method further comprises administering the proinflammatory agent locally (e.g., intratumorally) into the individual. In some embodiments, the method further comprises administering an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) to the individual (e.g., locally or systemically).In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered prior to (e.g., within about any of one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to) the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered sequentially (e.g., prior to or after the tyrosine kinase inhibitor and / or the proinflammatory agent). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered immediately after (e.g., within about any of 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours, after) the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the proinflammatory agent comprises an agent or is selected from the group consisting of R848, 3M-852A, motolimod, bromopindine, and visamminde. In some embodiments, the proinflammatory agent comprises a TLR agonist (e.g., R848) and a proinflammatory cytokine (e.g., IFN-g). In some embodiments, the TLR agonist is selected from the group consisting of LTA, CpG, Poly I;C, LPS, MPLA, flagellin, R848, visamminde, bromopindine, motolimod, and loxoribine.

[0082] In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and a proinflammatory agent (e.g., a TLR agonist, e.g., radiation therapy), and wherein the method comprises administering the tyrosine kinase inhibitor orally, intravenously, or subcutaneously, optionally wherein the tyrosine kinase inhibitor is administered intermittently. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual for at least two cycles, further optionally wherein the tyrosine kinase inhibitor is administered at least once in each cycle, and wherein each cycle has from about three days to about twenty days. In some embodiments, the tyrosine kinase inhibitor is administered at least twice (e.g., for at least two consecutive days) in each cycle. In some embodiments, the tyrosine kinase inhibitor is administered at least three times (e.g., for at least three consecutive days) in each cycle. In some embodiments, the tyrosine kinase inhibitor is administered concurrently with the proinflammatory agent. In some embodiments, the tyrosine kinase inhibitor is administered concurrently with the proinflammatory agent. In some embodiments, the tyrosine kinase inhibitor and the proinflammatory agent are administered sequentially within 2 weeks (e.g., within 10 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or the same day). In some embodiments, the tyrosine kinase inhibitor has a half-life of no more than about 10 days (e.g., no more than about 7 days, 5 days, 4 days, or 3 days). In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of a small molecule, a nucleic acid (e.g., siRNA, shRNA, antisense RNA, microRNA), a nucleic acid editing system (e.g., a CRISPR system), and a protein agent (e.g., an antibody agent that targets a tyrosine kinase or an activated tyrosine kinase). In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling. In some embodiments, the tyrosine kinase inhibitor inhibits Src family kinases (SFKs). In some embodiments, the tyrosine kinase inhibitor is an Hck inhibitor. In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406. In some embodiments, the method further comprises administering the proinflammatory agent locally (e.g., intratumorally) to the individual. In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof). In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody).In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered prior to (e.g., within about any of one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to) the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered sequentially (e.g., prior to or after the tyrosine kinase inhibitor and / or the proinflammatory agent). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered immediately after (e.g., within about any of 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours, after) the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the proinflammatory agent comprises an agent or is selected from the group consisting of R848, 3M-852A, motolimod, bromopindine, and visamminde. In some embodiments, the proinflammatory agent comprises a TLR agonist (e.g., R848) and a proinflammatory cytokine (e.g., IFN-γ). In some embodiments, the TLR agonist is selected from the group consisting of LTA, CpG, Poly I;C, LPS, MPLA, flagellin, R848, visamminde, bromopindine, motolimod, and loxoribine.

[0083] In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising orally, intravenously, subcutaneously, and / or intratumorally administering to the individual a tyrosine kinase inhibitor and a proinflammatory agent (e.g., a TLR agonist, e.g., radiation therapy), optionally wherein the tyrosine kinase inhibitor is effective to inhibit more than 50% of tyrosine kinase activity for no more than about 5 days, and optionally wherein the method comprises administering the tyrosine kinase inhibitor to the individual at an interval of no more than once every three days at least two times (e.g., at least 3 times, 4 times, 5 times, or 6 times). In some embodiments, the tyrosine kinase inhibitor is administered intermittently. In some embodiments, the tyrosine kinase inhibitor is administered at an interval of no more than two times every seven to twenty days. In some embodiments, the tyrosine kinase inhibitor is administered at an interval of no more than three times every seven to twenty days. In some embodiments, the tyrosine kinase inhibitor is administered at an interval of about 1-3 times every seven to twenty days for a period of at least fourteen to twenty days. In some embodiments, the tyrosine kinase inhibitor is administered at least about 2 times, 3 times, 4 times, 5 times, or 6 times over a period of about fourteen to about forty days (e.g., about fourteen to about twenty days). In some embodiments, the tyrosine kinase inhibitor is administered concurrently with the proinflammatory agent. In some embodiments, the tyrosine kinase inhibitor is administered concurrently with the proinflammatory agent. In some embodiments, the tyrosine kinase inhibitor and the proinflammatory agent are administered sequentially within 2 weeks (e.g., within 10 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or the same day). In some embodiments, the tyrosine kinase inhibitor has a half-life of no more than about 10 days (e.g., no more than about 7 days, 5 days, 4 days, or 3 days). In some embodiments, the tyrosine kinase inhibitor is effective to inhibit more than 50% of tyrosine kinase activity for no more than about 7 days (e.g., about 5 days, 4 days, or 3 days). In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of a small molecule, a nucleic acid (e.g., siRNA, shRNA, antisense RNA, microRNA), a nucleic acid editing system (e.g., a CRISPR system), and a protein agent (e.g., an antibody agent that targets a tyrosine kinase or an activated tyrosine kinase). In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling. In some embodiments, the tyrosine kinase inhibitor inhibits a Src family kinase (SFK). In some embodiments, the tyrosine kinase inhibitor is an Hck inhibitor. In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406. In some embodiments, the method further comprises locally (e.g., intratumorally) administering the proinflammatory agent to the individual. In some embodiments, the tyrosine kinase inhibitor is administered systemically and the proinflammatory agent is administered intratumorally.In some embodiments, the tyrosine kinase inhibitor is administered systemically and intratumorally. In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof). In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered prior to (e.g., within about any of one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to) the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered sequentially (e.g., prior to or after) the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered immediately after (e.g., within about any of 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours, after) the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the proinflammatory agent comprises an agent or is selected from the group consisting of R848, 3M-852A, motolimod, bromopindine, and visamminde. In some embodiments, the proinflammatory agent comprises a TLR agonist (e.g., R848) and a proinflammatory cytokine (e.g., IFN-g). In some embodiments, the TLR agonist is selected from the group consisting of LTA, CpG, Poly I;C, LPS, MPLA, flagellin, R848, visamminde, bromopindine, motolimod, and loxoribine.

[0084] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising orally, intravenously, subcutaneously, and / or intratumorally administering to the individual a tyrosine kinase inhibitor and a proinflammatory agent (e.g., a TLR agonist, e.g., radiation therapy), wherein the tyrosine kinase inhibitor is effective to inhibit more than 50% of tyrosine kinase activity for no more than about 5 days (e.g., no more than 5 days, 4 days, or 3 days). In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual for at least two cycles, wherein the tyrosine kinase inhibitor is administered at least once in each cycle, and wherein each cycle has from about three days to about twenty days. In some embodiments, the tyrosine kinase inhibitor is administered at least twice (e.g., at least for two consecutive days) in each cycle. In some embodiments, the tyrosine kinase inhibitor is administered at least three times (e.g., at least for three consecutive days) in each cycle. In some embodiments, the tyrosine kinase inhibitor is administered concurrently with the proinflammatory agent. In some embodiments, the tyrosine kinase inhibitor is administered concurrently with the proinflammatory agent. In some embodiments, the tyrosine kinase inhibitor and the proinflammatory agent are administered sequentially within 2 weeks (e.g., within 10 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or the same day). In some embodiments, the tyrosine kinase inhibitor has a half-life of no more than about 10 days (e.g., no more than about 7 days, 5 days, 4 days, or 3 days). In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of a small molecule, a nucleic acid (e.g., siRNA, shRNA, antisense RNA, microRNA), a nucleic acid editing system (e.g., a CRISPR system), and a protein agent (e.g., an antibody agent that targets a tyrosine kinase or an activated tyrosine kinase). In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling. In some embodiments, the tyrosine kinase inhibitor inhibits Src family kinases (SFKs). In some embodiments, the tyrosine kinase inhibitor is an Hck inhibitor. In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406. In some embodiments, the method further comprises locally (e.g., intratumorally) administering the proinflammatory agent to the individual. In some embodiments, the tyrosine kinase inhibitor is administered systemically and the proinflammatory agent is administered intratumorally. In some embodiments, the tyrosine kinase inhibitor is administered systemically and intratumorally. In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof). In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody).In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered prior to (e.g., within about any of one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to) the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered sequentially (e.g., prior to or after the tyrosine kinase inhibitor and / or the proinflammatory agent). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered immediately after (e.g., within about any of 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours, after) the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the proinflammatory agent comprises an agent or is selected from the group consisting of R848, 3M-852A, motolimod, bromopindine, and visamminde. In some embodiments, the proinflammatory agent comprises a TLR agonist (e.g., R848) and a proinflammatory cytokine (e.g., IFN-γ). In some embodiments, the TLR agonist is selected from the group consisting of LTA, CpG, Poly I;C, LPS, MPLA, flagellin, R848, visamminde, bromopindine, motolimod, and loxoribine.

[0085] In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual (e.g., orally, intravenously, subcutaneously, and / or intratumorally) a tyrosine kinase inhibitor and an immune cell, such as any of the immune cells described herein. In some embodiments, the individual has received, is receiving, or will receive a proinflammatory agent (e.g., a TLR agonist, e.g., R848, e.g., radiation therapy). In some embodiments, the individual is in an inflammatory response or has an ongoing infection. In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual (e.g., intravenously, subcutaneously, and / or intratumorally) a tyrosine kinase inhibitor and a proinflammatory agent (e.g., a TLR agonist, e.g., radiation therapy) and an immune cell. In some embodiments, the immune cell is derived from the same individual. In some embodiments, the immune cell comprises a monocyte or macrophage. In some embodiments, the immune cell comprises a T cell (e.g., a CAR-T cell). In some embodiments, the immune cell comprises an NK cell (e.g., a CAR-NK cell). In some embodiments, the immune cell comprises a neutrophil (e.g., a CAR-expressing neutrophil). In some embodiments, the immune cell comprises an antigen presenting cell (APC). In some embodiments, the immune cell is engineered to express a chimeric receptor that specifically binds to a tumor antigen. In some embodiments, the tyrosine kinase inhibitor is administered intermittently. In some embodiments, the tyrosine kinase inhibitor, immune cell, and / or proinflammatory agent are administered within 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or 1 day of each other. In some embodiments, the tyrosine kinase inhibitor and immune cell are administered within 24 hours of each other (e.g., within 12 hours, 8 hours, 4 hours, 2 hours, or 1 hour of each other, or within 30 minutes of each other). In some embodiments, the tyrosine kinase inhibitor, immune cell, and / or proinflammatory agent are administered simultaneously. In some embodiments, the tyrosine kinase inhibitor, immune cell, and / or proinflammatory agent are administered concurrently. In some embodiments, the tyrosine kinase inhibitor, immune cell, and / or proinflammatory agent are administered sequentially. In some embodiments, the tyrosine kinase inhibitor is administered systemically and the proinflammatory agent is administered intratumorally. In some embodiments, the tyrosine kinase inhibitor is administered systemically and intratumorally. In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling. In some embodiments, the tyrosine kinase inhibitor inhibits a Src family kinase (SFK). In some embodiments, the tyrosine kinase inhibitor is an Hck inhibitor. In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES.In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406. In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof). In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered prior to (e.g., within about any of one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to) the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered sequentially (e.g., prior to or after) the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered immediately after (e.g., within about any of 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours, after) the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the proinflammatory agent comprises an agent or is selected from the group consisting of R848, 3M-852A, motolimod, bromopindine, and visamminde. In some embodiments, the proinflammatory agent comprises a TLR agonist (e.g., R848) and a proinflammatory cytokine (e.g., IFN-g). In some embodiments, the TLR agonist is selected from the group consisting of LTA, CpG, Poly I;C, LPS, MPLA, flagellin, R848, visamminde, bromopindine, motolimod, and losoxantrone.

[0086] In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and a TLR agonist (e.g., R848), wherein the tyrosine kinase inhibitor is administered at least twice (e.g., at least 3, 4, or 5 times). In some embodiments, the tyrosine kinase inhibitor is administered intermittently. In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and a TLR agonist, wherein the tyrosine kinase inhibitor and the TLR agonist are administered within 24 hours of each other (e.g., within 12 hours, 8 hours, 4 hours, 2 hours, or 1 hour of each other, or within 30 minutes of each other). In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual at least twice at intervals of no more than once every three days. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual for at least two cycles, wherein the tyrosine kinase inhibitor is administered at least once (e.g., at least two or three times) in each cycle, and wherein each cycle has a duration of about three days to about twenty days. In some embodiments, the tyrosine kinase inhibitor is administered systemically (e.g., intravenously or subcutaneously) and / or locally (e.g., intratumorally). In some embodiments, the tyrosine kinase inhibitor and the TLR agonist are administered simultaneously, concurrently, or sequentially. In some embodiments, the TLR agonist activates TLR1 or TLR2, optionally wherein the TLR agonist comprises triacylated lipoprotein, peptidoglycan, zymosan, and / or Pam3CSK4. In some embodiments, the TLR agonist activates any of TLR2, TLR3, TLR4, TLR5, and TLR6, optionally wherein the TLR agonist comprises diacylated lipopeptide, heat shock protein, HMGB1, uric acid, fibronectin, and / or ECM protein. In some embodiments, the TLR agonist activates TLR2, optionally wherein the TLR agonist comprises Pam3Cys, SMP-105, and / or CBLB612. In some embodiments, the TLR agonist activates TLR3, optionally wherein the TLR agonist comprises dsRNA, Poly I:C, PolyICIC, Poly-IC12U, IPH302, ARNAX, and / or MPLA. In some embodiments, the TLR agonist activates TLR4, optionally wherein the TLR agonist comprises LPS, lipoteichoic acid beta-defensin 2, fibronectin EDA, HMGB1, snapin, tenascin C, OK-432, AS04, and / or GLA-SE. In some embodiments, the TLR agonist activates TLR5, optionally wherein the TLR agonist comprises flagellin, CBLB502, and / or M-VM3. In some embodiments, the TLR agonist activates TLR6.In some embodiments, the TLR agonist activates TLR7 or TLR8, optionally wherein the TLR agonist comprises ssRNA, CpG-A, poly G10, and / or poly G3. In some embodiments, the TLR agonist activates TLR7, optionally wherein the TLR agonist comprises bistriazolyl and / or R848. In some embodiments, the TLR agonist activates TLR8, optionally wherein the TLR agonist comprises VTX1463 and / or R848. In some embodiments, the TLR agonist activates TLR9, optionally wherein the TLR agonist comprises unmethylated CpG DNA, CpG (e.g., CpG-7909, KSK-CpG, CpG-1826), MGN1703, dsSLIM, IMO2055, SD101, and / or ODN M362. In some embodiments, the TLR agonist activates TLR10, optionally wherein the TLR agonist comprises Pam3CSK4. In some embodiments, the TLR agonist activates TLR11, optionally wherein the TLR agonist comprises Toxoplasma gondii inhibitor protein. In some embodiments, the TLR agonist activates TLR12. In some embodiments, the TLR agonist activates TLR13, optionally wherein the TLR agonist comprises VSV. In some embodiments, the TLR agonist activates TLR1, TLR2, TLR3, TLR4, TLR7, TLR8, and / or TLR9. In some embodiments, the TLR agonist activates TLR9, TLR4, and TLR7 / 8. In some embodiments, the TLR agonist comprises CpG, poly I:C, and / or R848. In some embodiments, the TLR agonist comprises CpG, poly I:C, and R848, e.g., at a ratio of 1:1:1. In some embodiments, the tyrosine kinase inhibitor is administered systemically and the TLR agonist is administered intratumorally. In some embodiments, the tyrosine kinase inhibitor is administered systemically and intratumorally. In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling. In some embodiments, the tyrosine kinase inhibitor inhibits Src family kinases (SFKs). In some embodiments, the tyrosine kinase inhibitor is an Hck inhibitor. In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406. In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof). In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody).In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered prior to (e.g., within about any of one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to) the tyrosine kinase inhibitor and / or TLR agonist. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or TLR agonist. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or TLR agonist. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered sequentially (e.g., prior to or after the tyrosine kinase inhibitor and / or TLR agonist). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered immediately after (e.g., within about any of 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours, after) the tyrosine kinase inhibitor and / or TLR agonist. In some embodiments, the TLR agonist is selected from the group consisting of LTA, CpG, Poly I;C, LPS, MPLA, flagellin, R848, vesatmold, bromopindine, motolimod, and losoxantrone.

[0087] In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering a tyrosine kinase inhibitor and a TLR agonist (e.g., R848), optionally wherein the TLR agonist activates one or more TLRs selected from the group consisting of TLR9, TLR4, TLR7, and TLR8. In some embodiments, the tyrosine kinase inhibitor and the TLR agonist are administered within the same day. In some embodiments, the tyrosine kinase inhibitor is administered intermittently. In some embodiments, the tyrosine kinase inhibitor and / or the TLR agonist is administered at least twice (e.g., at least three, four, five, or six times). In some embodiments, the tyrosine kinase inhibitor and the TLR agonist are administered for at least two cycles (e.g., at least three cycles), optionally wherein the tyrosine kinase inhibitor and the TLR agonist are administered within the same day for at least two consecutive days (e.g., at least three consecutive days) in each cycle. In some embodiments, each cycle has from about seven days to about twenty days. In some embodiments, the TLR agonist activates a TLR on a macrophage, optionally wherein the TLR comprises TLR9. In some embodiments, the TLR agonist activates at least two TLRs (e.g., TLR4, TLR7, TLR8, or TLR9). In some embodiments, the TLR agonist activates at least three TLRs (e.g., TLR9, TLR4, and TLR7 / 8). In some embodiments, the TLR agonist comprises CpG, poly I:C, and / or R848. In some embodiments, the TLR agonist comprises CpG, poly I:C, and R848, e.g., at a ratio of 1 : 1 : 1. In some embodiments, the tyrosine kinase inhibitor is administered systemically, and the TLR agonist is administered intratumorally. In some embodiments, the tyrosine kinase inhibitor is administered systemically and intratumorally. In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling. In some embodiments, the tyrosine kinase inhibitor inhibits Src family kinases (SFKs). In some embodiments, the tyrosine kinase inhibitor is an Hck inhibitor. In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406. In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof). In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody).In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered prior to (e.g., within about any of one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to) the tyrosine kinase inhibitor and / or TLR agonist. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or TLR agonist. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or TLR agonist. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered sequentially (e.g., prior to or after the tyrosine kinase inhibitor and / or TLR agonist). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered immediately after (e.g., within about any of 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours, after) the tyrosine kinase inhibitor and / or TLR agonist. In some embodiments, the TLR agonist is selected from the group consisting of LTA, CpG, Poly I;C, LPS, MPLA, flagellin, R848, vesatmold, bromopindine, motolimod, and losoxantrone.

[0088] In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and a STING activator (e.g., MSA-2, ADU-S100, or cGAMP), optionally wherein the tyrosine kinase inhibitor is administered at least twice (at least three, four, five, or six times). In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and a STING activator (e.g., MSA-2, ADU-S100, or cGAMP), optionally wherein the tyrosine kinase inhibitor and the STING activator are administered within 24 hours of each other (e.g., within 12 hours, 8 hours, 4 hours, 2 hours, or 1 hour of each other, or within 30 minutes of each other). In some embodiments, the tyrosine kinase inhibitor is administered intermittently. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual at least twice at intervals of no more than once every three days. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual for at least two cycles, wherein the tyrosine kinase inhibitor is administered at least once in each cycle, and wherein each cycle has about three days to about twenty days. In some embodiments, the tyrosine kinase inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or locally (e.g., intratumorally). In some embodiments, the tyrosine kinase inhibitor and the STING activator are administered sequentially, simultaneously, or concurrently. In some embodiments, the STING activator is a cyclic guanosine monophosphate-adenosine monophosphate (cGAMP, e.g., 3'3' cGAMP, e.g., 2'3' cGAMP), a bacterial vector (e.g., SYNB1891, STACT-TREX-1), a CDN compound (e.g., ADU-S100, BI-STING, BMS-986301, GSK532, JNJ-4412, MK-1454, SB11285, 3'3'-cyclic AIMP), a non-CDN small molecule (e.g., ALG-031048, E7755, JNJ-‘6196, MK-2118, MSA-1, MSA-2, SNX281, SR-717, TAK676, TT I-10001), a nanovaccine (e.g., PC7A NP, cCAMP-NP, ONM-500), or an antibody-drug conjugate (e.g., XMT-2056, CRD-5500). In some embodiments, the tyrosine kinase inhibitor is administered systemically, and the STING activator is administered intratumorally. In some embodiments, the tyrosine kinase inhibitor is administered systemically and intratumorally. In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling.In some embodiments, the tyrosine kinase inhibitor inhibits a Src family kinase (SFK). In some embodiments, the tyrosine kinase inhibitor is an Hck inhibitor. In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406. In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof). In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody). In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered prior to (e.g., within any of about one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to) the tyrosine kinase inhibitor and / or the STING activator. In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the STING activator. In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the STING activator. In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered sequentially (e.g., prior to or after) the tyrosine kinase inhibitor and / or the STING activator. In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered immediately after (e.g., within any of about 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours, after) the tyrosine kinase inhibitor and / or the STING activator.

[0089] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and radiation therapy, optionally wherein the method comprises administering the tyrosine kinase inhibitor to the individual for at least two cycles, wherein the tyrosine kinase inhibitor is administered at least once per cycle, and wherein each cycle has from about three days to about twenty days. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual at least twice at intervals of no more than once every three days. In some embodiments, the tyrosine kinase inhibitor is administered at least three times per day. In some embodiments, the tyrosine kinase inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or locally (e.g., intratumorally). In some embodiments, the tyrosine kinase inhibitor and the radiation therapy are administered within 24 hours of each other (e.g., within 12 hours, 8 hours, 4 hours, 2 hours, or 1 hour of each other, or within 30 minutes of each other). In some embodiments, the radiation therapy comprises irradiation at a site of the cancer to be treated. In some embodiments, the radiation therapy comprises irradiation at a site different from a site of the cancer to be treated. In some embodiments, the dose of the radiation therapy is insufficient to kill tumor cells. In some embodiments, the radiation therapy is selected from the group consisting of external beam radiation therapy, internal radiation therapy (brachytherapy), intraoperative radiation therapy (IORT), whole body radiation therapy, radioimmunotherapy, and administration of radiosensitizers and radioprotectors. In some embodiments, the radiation therapy is external beam radiation therapy, optionally comprising three-dimensional conformal radiation therapy (3D-RT), intensity modulated radiation therapy (IMRT), photon beam therapy, image-guided radiation therapy (IGRT), and stereotactic radiation therapy (SRT). In some embodiments, the radiation therapy is brachytherapy, optionally comprising interstitial brachytherapy, intracavitary brachytherapy, intraluminal brachytherapy, and intravenously administered radiolabeled molecules. In some embodiments, the tyrosine kinase inhibitor is administered systemically and intratumorally. In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling. In some embodiments, the tyrosine kinase inhibitor inhibits Src family kinases (SFKs). In some embodiments, the tyrosine kinase inhibitor is an Hck inhibitor. In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406. In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof).In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody). In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered prior to (e.g., within about any of one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to) the tyrosine kinase inhibitor and / or the radiation therapy. In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the radiation therapy. In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the radiation therapy. In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered sequentially (e.g., prior to or after) the tyrosine kinase inhibitor and / or the radiation therapy. In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered immediately after (e.g., within about any of 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours, after) the tyrosine kinase inhibitor and / or the radiation therapy.

[0090] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and radiation therapy, wherein the radiation therapy comprises irradiation at a site different from the site of the cancer to be treated. In some embodiments, the tyrosine kinase inhibitor is administered at least two times (at least three, four, five, or six times). In some embodiments, the tyrosine kinase inhibitor is administered intermittently. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual at least two times at intervals of no more than once every three days. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual for at least two cycles, wherein the tyrosine kinase inhibitor is administered at least once in each cycle, and wherein each cycle has about three days to about twenty days. In some embodiments, the tyrosine kinase inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or locally (e.g., intratumorally). In some embodiments, the tyrosine kinase inhibitor and the radiation therapy are administered within 24 hours of each other (e.g., within 12 hours, 8 hours, 4 hours, 2 hours, or 1 hour of each other, or within 30 minutes of each other). In some embodiments, the radiation therapy comprises irradiation at the site of the cancer to be treated. In some embodiments, the radiation therapy comprises irradiation at a site different from the site of the cancer to be treated. In some embodiments, the dose of the radiation therapy is insufficient to kill tumor cells. In some embodiments, the radiation therapy is selected from the group consisting of external beam radiation therapy, internal radiation therapy (brachytherapy), intraoperative radiation therapy (IORT), whole body radiation therapy, radioimmunotherapy, and administration of radiosensitizers and radioprotectors. In some embodiments, the radiation therapy is external beam radiation therapy, optionally comprising three-dimensional conformal radiation therapy (3D-RT), intensity modulated radiation therapy (IMRT), photon beam therapy, image-guided radiation therapy (IGRT), and stereotactic radiation therapy (SRT). In some embodiments, the radiation therapy is brachytherapy, optionally comprising interstitial brachytherapy, intracavitary brachytherapy, intraluminal brachytherapy, and intravenously administered radiolabeled molecules. In some embodiments, the tyrosine kinase inhibitor is administered systemically and intratumorally. In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling. In some embodiments, the tyrosine kinase inhibitor inhibits Src family kinases (SFKs). In some embodiments, the tyrosine kinase inhibitor is an Hck inhibitor. In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406.In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof). In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered prior to (e.g., within about any of one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to) the tyrosine kinase inhibitor and / or radiation therapy. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or radiation therapy. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or radiation therapy. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered sequentially (e.g., prior to or after the tyrosine kinase inhibitor and / or radiation therapy). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered immediately after (e.g., within about any of 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours, after) the tyrosine kinase inhibitor and / or radiation therapy.

[0091] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering a tyrosine kinase inhibitor and radiation therapy. In some embodiments, the tyrosine kinase inhibitor is administered intermittently. In some embodiments, the tyrosine kinase inhibitor and radiation therapy are administered within the same day. In some embodiments, the tyrosine kinase inhibitor and / or radiation therapy is administered at least two times (e.g., at least three, four, five, or six times). In some embodiments, the tyrosine kinase inhibitor and radiation therapy are administered for at least two cycles (e.g., at least three cycles), optionally wherein the tyrosine kinase inhibitor and radiation therapy are administered within the same day in each cycle for at least two consecutive days (e.g., at least three consecutive days). In some embodiments, each cycle has about seven days to about twenty days. In some embodiments, the tyrosine kinase inhibitor and radiation therapy are administered within 24 hours of each other (e.g., within 12 hours, 8 hours, 4 hours, 2 hours, or 1 hour of each other, or within 30 minutes of each other). In some embodiments, the radiation therapy comprises irradiation at a site of a cancer to be treated. In some embodiments, the radiation therapy comprises irradiation at a site different from a site of a cancer to be treated. In some embodiments, the dose of radiation therapy is insufficient to kill tumor cells. In some embodiments, the tyrosine kinase inhibitor is administered systemically and intratumorally. In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling. In some embodiments, the tyrosine kinase inhibitor inhibits Src family kinases (SFKs). In some embodiments, the tyrosine kinase inhibitor is an Hck inhibitor. In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406. In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof). In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody). In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered prior to the tyrosine kinase inhibitor and / or radiation therapy (e.g., within about any of one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to the tyrosine kinase inhibitor and / or radiation therapy).In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered concurrently with the tyrosine kinase inhibitor and / or radiation therapy. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered concurrently with the tyrosine kinase inhibitor and / or radiation therapy. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered sequentially (e.g., prior to or after the tyrosine kinase inhibitor and / or radiation therapy). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered immediately after the tyrosine kinase inhibitor and / or radiation therapy (e.g., within any of about 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours after the tyrosine kinase inhibitor and / or radiation therapy).

[0092] In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and a PAMP / DAMP activator, optionally wherein the tyrosine kinase inhibitor is administered at least twice (at least three, four, five, or six times). In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and a PAMP / DAMP activator, optionally wherein the tyrosine kinase inhibitor and the PAMP / DAMP activator are administered within 24 hours of each other (e.g., within 12 hours, 8 hours, 4 hours, 2 hours, or 1 hour of each other, or within 30 minutes of each other). In some embodiments, the tyrosine kinase inhibitor is administered intermittently. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual at least twice at an interval of no more than once every three days. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual for at least two cycles, wherein the tyrosine kinase inhibitor is administered at least once in each cycle, and wherein each cycle has about three days to about twenty days. In some embodiments, the tyrosine kinase inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or locally (e.g., intratumorally). In some embodiments, the proinflammatory agent is a PAMP activator. In some embodiments, the PAMP activator is a triacyl lipopeptide, LPS, lipoprotein, peptidoglycan, zymosan, lipoteichoic acid, trypanosomal phospholipid, Pam3Cys porin, lipoarabinomannan, double stranded RNA, poly(l:C), trypanosomal lipid, pseudomonas exoenzyme S, RSV F protein, MMTV envelope protein, flagellin, diacyl lipopeptide, single stranded RNA, imiquimod, single stranded RNA, resiquimod, bacterial / viral DNA, CpG DNA, urea bacterial, or toxoplasma LPS. In some embodiments, the proinflammatory agent is a DAMP activator. In some embodiments, the DAMP activator is a defensin, HSP60, HSP70, messenger RNA, low molecular weight hyaluronan, fibrinogen, fibronectin, fx1-defensin, heparan sulfate, HSP60, HSP70, HSP90, HMGB1, or unmethylated CpG DNA. In some embodiments, the tyrosine kinase inhibitor is administered systemically and the PAMP / DAMP activator is administered intratumorally. In some embodiments, the tyrosine kinase inhibitor is administered systemically and intratumorally. In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling. In some embodiments, the tyrosine kinase inhibitor inhibits Src family kinases (SFKs). In some embodiments, the tyrosine kinase inhibitor is an Hck inhibitor.In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406. In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof). In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered prior to (e.g., within any of about one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to) the tyrosine kinase inhibitor and / or the PAMP / DAMP activator. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the PAMP / DAMP activator. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the PAMP / DAMP activator. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered sequentially (e.g., prior to or after) the tyrosine kinase inhibitor and / or the PAMP / DAMP activator. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered immediately after (e.g., within any of about 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours, after) the tyrosine kinase inhibitor and / or the PAMP / DAMP activator.

[0093] In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and a checkpoint inhibitor (e.g., an anti-PD-1 agent, an anti-PD-L1 agent, or an anti-CTLA-4 agent), optionally wherein the tyrosine kinase inhibitor is administered at least twice (at least three, four, five, or six times). In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and a checkpoint inhibitor (e.g., an anti-PD-1 agent, an anti-PD-L1 agent, or an anti-CTLA-4 agent), wherein the tyrosine kinase inhibitor and the checkpoint inhibitor are administered within 24 hours of each other (e.g., within 12 hours, 8 hours, 4 hours, 2 hours, or 1 hour of each other, or within 30 minutes of each other). In some embodiments, the tyrosine kinase inhibitor is administered intermittently. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual at least twice at an interval of no more than once every three days. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual for at least two cycles, wherein the tyrosine kinase inhibitor is administered at least once in each cycle, and wherein each cycle has about three days to about twenty days. In some embodiments, the tyrosine kinase inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or locally (e.g., intratumorally). In some embodiments, the checkpoint inhibitor targets LAG-3, TIM-3, B7-H3, B7-H4, A2aR, CD73, NKG2A, PVRIG / PVRL2, CEACAM1, CEACAM 5 / 6, FAK, CCL2 / CCR2, LIF, CD47 / SIRPalpha, CSF-1 (M-CSF) / CSF-1R, IL-1 / IL-1R3 (IL-1RAP), IL-8, SEMA4D, Ang-2, CLEVER-1, Axl, or phosphatidylserine.In some embodiments, the checkpoint inhibitor comprises or is Libivimab, Cemiplimab, Nivolumab, Pembrolizumab, Atezolizumab, Avelumab, Durvalumab, LAG525 (IMP701), REGN 3767, BI 754,091, Toripalimab (MGD013), Ipilimumab (IMP321), FS118, MBG453, Sym023, TSR-022, MGC018, FPA150, EOS100850, AB928, CPI-006, Monalizumab, COM701, CM24, NEO-201, Defactinib, PF-04136309, MSC-1, Hu5F9-G4 (5F9), ALX148, TTI-662, RRx-001, Lannotuzumab (MCS110), LY3022855, SNDX-6352, Ensituximab (RG7155), Plitharanthine (PLX3397), CAN04, Canakinumab (ACZ885), BMS-986253, Pembinafumab (VX15 / 2503), Tremikimab, FP-1305, Vincitutumumab (EnaV), or Batiximab. In some embodiments, the tyrosine kinase inhibitor is administered systemically and the checkpoint inhibitor is administered intratumorally. In some embodiments, the tyrosine kinase inhibitor is administered systemically and intratumorally. In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling. In some embodiments, the tyrosine kinase inhibitor inhibits Src family kinases (SFKs). In some embodiments, the tyrosine kinase inhibitor is a Hck inhibitor. In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406. In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof). In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered prior to (e.g., within about any one of one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to) the tyrosine kinase inhibitor and / or the checkpoint inhibitor.In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered simultaneously with the tyrosine kinase inhibitor and / or checkpoint inhibitor. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or checkpoint inhibitor. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered sequentially (e.g., before or after the tyrosine kinase inhibitor and / or checkpoint inhibitor). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered immediately after the tyrosine kinase inhibitor and / or checkpoint inhibitor (e.g., within any of about 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours after the tyrosine kinase inhibitor and / or checkpoint inhibitor).

[0094] In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and a proinflammatory cytokine (e.g., IL-1b, IL-18, IL-6, and / or TNFa), optionally wherein the tyrosine kinase inhibitor is administered at least twice (at least three, four, five, or six times). In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and a proinflammatory cytokine (e.g., IL-1b, IL-18, IL-6, and / or TNFa), wherein the tyrosine kinase inhibitor and the proinflammatory cytokine are administered within 24 hours of each other (e.g., within 12 hours, 8 hours, 4 hours, 2 hours, or 1 hour of each other, or within 30 minutes of each other). In some embodiments, the tyrosine kinase inhibitor is administered intermittently. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual at least twice at an interval of no more than once every three days. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual for at least two cycles, wherein the tyrosine kinase inhibitor is administered at least once in each cycle, and wherein each cycle has about three days to about twenty days. In some embodiments, the tyrosine kinase inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or locally (e.g., intratumorally). In some embodiments, the proinflammatory cytokine promotes M1 macrophages. In some embodiments, the proinflammatory cytokine comprises or is TNF, IFNy, and / or GM-CSF. In some embodiments, the proinflammatory cytokine comprises IFNy. In some embodiments, the proinflammatory cytokine comprises IL-1. In some embodiments, the proinflammatory cytokine comprises TNF-a. In some embodiments, the proinflammatory cytokine comprises IL-6. In some embodiments, the tyrosine kinase inhibitor is administered systemically, and the proinflammatory cytokine is administered intratumorally. In some embodiments, the tyrosine kinase inhibitor is administered systemically and intratumorally. In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling. In some embodiments, the tyrosine kinase inhibitor inhibits Src family kinases (SFKs). In some embodiments, the tyrosine kinase inhibitor is an Hck inhibitor. In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406. In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof).In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered prior to (e.g., within about any of one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to) the tyrosine kinase inhibitor and / or the proinflammatory cytokine. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the proinflammatory cytokine. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the proinflammatory cytokine. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered sequentially (e.g., prior to or after) the tyrosine kinase inhibitor and / or the proinflammatory cytokine. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered immediately after (e.g., within about any of 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours, after) the tyrosine kinase inhibitor and / or the proinflammatory cytokine.

[0095] In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and a chemotherapeutic agent (e.g., azathioprine), optionally wherein the tyrosine kinase inhibitor is administered at least two (at least three, four, five, or six) times. In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and a chemotherapeutic agent (e.g., azathioprine), wherein the tyrosine kinase inhibitor and the chemotherapeutic agent are administered within 24 hours of each other (e.g., within 12 hours, 8 hours, 4 hours, 2 hours, or 1 hour of each other, or within 30 minutes of each other). In some embodiments, the tyrosine kinase inhibitor is administered intermittently. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual at least two times at an interval of no more than once every three days. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual for at least two cycles, wherein the tyrosine kinase inhibitor is administered at least once in each cycle, and wherein each cycle has about three days to about twenty days. In some embodiments, the tyrosine kinase inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or locally (e.g., intratumorally). In some embodiments, the chemotherapeutic agent is an alkylating agent. In some embodiments, the alkylating agent is selected from the group consisting of nitrogen mustards (e.g., endoxan, cyclophosphamide, ifosfamide), nitrosoureas (e.g., carmustine, lomustine), platinum analogs (e.g., carboplatin, cisplatin, oxaliplatin), triazenes (e.g., dacarbazine, procarbazine, temozolomide), alkyl sulfonates (e.g., busulfan), and ethylenimines (e.g., thiotepa). In some embodiments, the chemotherapeutic agent is an antimetabolite. In some embodiments, the antimetabolite is selected from the group consisting of cytidine analogs (e.g., azacitidine, decitabine, cytarabine, gemcitabine), folic acid antagonists (e.g., methotrexate, pemetrexed), purine analogs (e.g., cladribine, clofarabine, nelarabine), pyrimidine analogs (e.g., fluorouracil (5-FU), capecitabine (a prodrug of 5-FU)). In some embodiments, the chemotherapeutic agent is an anti-microtubule agent. In some embodiments, the anti-microtubule agent is selected from the group consisting of topoisomerase II inhibitors (e.g., anthracyclines, doxorubicin, daunorubicin, idarubicin, mitoxantrone), topoisomerase I inhibitors (e.g., irinotecan, topotecan), taxanes (e.g., paclitaxel, docetaxel, cabazitaxel), vinca alkaloids (e.g., vinblastine, vincristine, vinorelbine), antibiotics (e.g., actinomycin D, bleomycin, daunorubicin). In some embodiments, the chemotherapeutic agent is hydroxyurea, retinoid, arsenic trioxide, or a proteasome inhibitor (e.g., bortezomib).In some embodiments, the tyrosine kinase inhibitor is administered systemically and the chemotherapeutic agent is administered intratumorally. In some embodiments, the tyrosine kinase inhibitor is administered systemically and intratumorally. In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling. In some embodiments, the tyrosine kinase inhibitor inhibits Src family kinases (SFKs). In some embodiments, the tyrosine kinase inhibitor is an Hck inhibitor. In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406. In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof). In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered prior to (e.g., within any of about one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to) the tyrosine kinase inhibitor and / or the chemotherapeutic agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the chemotherapeutic agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the chemotherapeutic agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered sequentially (e.g., prior to or after) the tyrosine kinase inhibitor and / or the chemotherapeutic agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered immediately after (e.g., within any of about 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours, after) the tyrosine kinase inhibitor and / or the chemotherapeutic agent.

[0096] In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and a cancer vaccine, optionally wherein the tyrosine kinase inhibitor is administered at least twice. In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and a cancer vaccine, wherein the tyrosine kinase inhibitor and the cancer vaccine are administered within 24 hours of each other (e.g., within 12 hours, 8 hours, 4 hours, 2 hours, or 1 hour of each other, or within 30 minutes of each other). In some embodiments, the tyrosine kinase inhibitor is administered intermittently. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual at least twice at intervals of no more than once every three days. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual for at least two cycles, wherein the tyrosine kinase inhibitor is administered at least once in each cycle, and wherein each cycle has about three days to about twenty days. In some embodiments, the tyrosine kinase inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or locally (e.g., intratumorally). In some embodiments, the cancer vaccine comprises a cell-based vaccine, a peptide-based vaccine, a virus-based vaccine, and / or a nucleic acid-based vaccine. In some embodiments, the tyrosine kinase inhibitor is administered systemically and the cancer vaccine is administered intratumorally. In some embodiments, the tyrosine kinase inhibitor is administered systemically and intratumorally. In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling. In some embodiments, the tyrosine kinase inhibitor inhibits Src family kinases (SFKs). In some embodiments, the tyrosine kinase inhibitor is an Hck inhibitor. In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406. In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof). In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody). In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered prior to (e.g., within any of about one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to) the tyrosine kinase inhibitor and / or the cancer vaccine.In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or cancer vaccine. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or cancer vaccine. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered sequentially (e.g., prior to or after the tyrosine kinase inhibitor and / or cancer vaccine). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered immediately after the tyrosine kinase inhibitor and / or cancer vaccine, e.g., within any of about 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours after the tyrosine kinase inhibitor and / or cancer vaccine.

[0097] In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and an oncolytic virus, optionally wherein the tyrosine kinase inhibitor is administered at least two (at least three, four, five, or six) times. In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and an oncolytic virus, wherein the tyrosine kinase inhibitor and the oncolytic virus are administered within 24 hours of each other (e.g., within 12 hours, 8 hours, 4 hours, 2 hours, or 1 hour of each other, or within 30 minutes of each other). In some embodiments, the tyrosine kinase inhibitor is administered intermittently. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual at least two times at an interval of no more than once every three days. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual for at least two cycles, wherein the tyrosine kinase inhibitor is administered at least once in each cycle, and wherein each cycle has about three days to about twenty days. In some embodiments, the tyrosine kinase inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or locally (e.g., intratumorally). In some embodiments, the oncolytic virus comprises or is an adenovirus (e.g., ONYX-15, LOAd703 virus), a parvovirus, a parvovirus (e.g., H-1PV), a vaccinia virus (VACV), a reovirus (e.g., Reolysin), or a herpes simplex virus (HSV, e.g., HSV-1, HSV-2, G207, L1BR1, HF10, T-VEC, Orien X010). In some embodiments, the oncolytic virus comprises JX-593, Coxsackievirus A21 (CVA21), a Maraba virus or its MG1 variant, a DNX2440 adenovirus, a fowlpox virus, or a Sendai virus. In some embodiments, the tyrosine kinase inhibitor is administered systemically, and the oncolytic virus is administered intratumorally. In some embodiments, the tyrosine kinase inhibitor is administered systemically and intratumorally. In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling. In some embodiments, the tyrosine kinase inhibitor inhibits a Src family kinase (SFK). In some embodiments, the tyrosine kinase inhibitor is an Hck inhibitor. In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406. In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof).In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody). In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered prior to (e.g., within any of about one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to) the tyrosine kinase inhibitor and / or the oncolytic virus. In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the oncolytic virus. In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the oncolytic virus. In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered sequentially (e.g., prior to or after) the tyrosine kinase inhibitor and / or the oncolytic virus. In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered immediately after (e.g., within any of about 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours, after) the tyrosine kinase inhibitor and / or the oncolytic virus.

[0098] In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and a sound treatment (e.g., high intensity focused ultrasound (HIFU), e.g., low intensity focused ultrasound (LIPUS)), optionally wherein the tyrosine kinase inhibitor is administered at least two (at least three, four, five, or six) times. In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and a sound treatment (e.g., high intensity focused ultrasound (HIFU), e.g., low intensity focused ultrasound (LIPUS)), wherein the tyrosine kinase inhibitor and the sound treatment are administered within 24 hours of each other (e.g., within 12 hours, 8 hours, 4 hours, 2 hours, or 1 hour of each other, or within 30 minutes of each other). In some embodiments, the tyrosine kinase inhibitor is administered intermittently. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual at least two times at an interval of no more than once every three days. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual for at least two cycles, wherein the tyrosine kinase inhibitor is administered at least once in each cycle, and wherein each cycle has about three days to about twenty days. In some embodiments, the tyrosine kinase inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or locally (e.g., intratumorally). In some embodiments, the tyrosine kinase inhibitor is administered systemically, and the method comprises administering the sound treatment at the site of the cancer to be treated. In some embodiments, the tyrosine kinase inhibitor is administered systemically and intratumorally. In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling. In some embodiments, the tyrosine kinase inhibitor inhibits Src family kinases (SFKs). In some embodiments, the tyrosine kinase inhibitor is an Hck inhibitor. In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406. In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof). In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody).In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered prior to (e.g., within any of about one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to) the tyrosine kinase inhibitor and / or the sound treatment. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the sound treatment. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the sound treatment. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered sequentially (e.g., prior to or after the tyrosine kinase inhibitor and / or the sound treatment). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered immediately after (e.g., within any of about 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours, after) the tyrosine kinase inhibitor and / or the sound treatment.

[0099] In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and a magnetotherapy (e.g., a pulsed magnetic field, e.g., a static magnetic field), optionally wherein the tyrosine kinase inhibitor is administered at least twice (at least three, four, five, or six times). In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and a magnetotherapy (e.g., a pulsed magnetic field, e.g., a static magnetic field), wherein the tyrosine kinase inhibitor and the magnetotherapy are administered within 24 hours of each other (e.g., within 12 hours, 8 hours, 4 hours, 2 hours, or 1 hour of each other, or within 30 minutes of each other). In some embodiments, the tyrosine kinase inhibitor is administered intermittently. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual at least twice at an interval of no more than once every three days. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual for at least two cycles, wherein the tyrosine kinase inhibitor is administered at least once in each cycle, and wherein each cycle has about three days to about twenty days. In some embodiments, the tyrosine kinase inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or locally (e.g., intratumorally). In some embodiments, the tyrosine kinase inhibitor is administered systemically, and the method comprises administering the magnetotherapy at the site of the cancer to be treated. In some embodiments, the tyrosine kinase inhibitor is administered systemically and intratumorally. In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling. In some embodiments, the tyrosine kinase inhibitor inhibits Src family kinases (SFKs). In some embodiments, the tyrosine kinase inhibitor is an Hck inhibitor. In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406. In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof). In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody). In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered prior to (e.g., within any of about one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to) the tyrosine kinase inhibitor and / or the magnetotherapy.In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the magnetic therapy. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the magnetic therapy. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered sequentially (e.g., before or after the tyrosine kinase inhibitor and / or the magnetic therapy). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered immediately after the tyrosine kinase inhibitor and / or the magnetic therapy (e.g., within any of about 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours after the tyrosine kinase inhibitor and / or the magnetic therapy).

[0100] In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and an electro- or electrochemical therapy, optionally wherein the tyrosine kinase inhibitor is administered at least twice (at least three, four, five, or six times). In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and an electro- or electrochemical therapy, wherein the tyrosine kinase inhibitor and the electro- or electrochemical therapy are administered within 24 hours of each other (e.g., within 12 hours, 8 hours, 4 hours, 2 hours, or 1 hour of each other, or within 30 minutes of each other). In some embodiments, the tyrosine kinase inhibitor is administered intermittently. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual at least twice at an interval of no more than once every three days. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual for at least two cycles, wherein the tyrosine kinase inhibitor is administered at least once in each cycle, and wherein each cycle has about three days to about twenty days. In some embodiments, the tyrosine kinase inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or locally (e.g., intratumorally). In some embodiments, the tyrosine kinase inhibitor is administered systemically, and the method comprises administering the electro- or electrochemical therapy at the site of the cancer to be treated. In some embodiments, the tyrosine kinase inhibitor is administered systemically and intratumorally. In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling. In some embodiments, the tyrosine kinase inhibitor inhibits Src family kinases (SFKs). In some embodiments, the tyrosine kinase inhibitor is an Hck inhibitor. In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406. In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof). In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody). In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered prior to the tyrosine kinase inhibitor and / or the electro- or electrochemical therapy (e.g., within any of about one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to the tyrosine kinase inhibitor and / or the electro- or electrochemical therapy).In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered simultaneously with the tyrosine kinase inhibitor and / or the electrical or electrochemical treatment. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the electrical or electrochemical treatment. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered sequentially (e.g., prior to or after the tyrosine kinase inhibitor and / or the electrical or electrochemical treatment). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered immediately after the tyrosine kinase inhibitor and / or the electrical or electrochemical treatment (e.g., within any of about 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours after the tyrosine kinase inhibitor and / or the electrical or electrochemical treatment).

[0101] In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and electrostatic therapy, optionally wherein the tyrosine kinase inhibitor is administered at least two times (at least three, four, five, or six times). In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor and electrostatic therapy, wherein the tyrosine kinase inhibitor and electrostatic therapy are administered within 24 hours of each other (e.g., within 12 hours, 8 hours, 4 hours, 2 hours, or 1 hour of each other, or within 30 minutes of each other). In some embodiments, the tyrosine kinase inhibitor is administered intermittently. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual at least two times at an interval of no more than once every three days. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual for at least two cycles, wherein the tyrosine kinase inhibitor is administered at least once in each cycle, and wherein each cycle has about three days to about twenty days. In some embodiments, the tyrosine kinase inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or locally (e.g., intratumorally). In some embodiments, the tyrosine kinase inhibitor is administered systemically, and the method comprises administering the electrostatic therapy at the site of the cancer to be treated. In some embodiments, the tyrosine kinase inhibitor is administered systemically and intratumorally. In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling. In some embodiments, the tyrosine kinase inhibitor inhibits Src family kinases (SFKs). In some embodiments, the tyrosine kinase inhibitor is an Hck inhibitor. In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406. In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof). In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody). In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered prior to the tyrosine kinase inhibitor and / or the electrostatic therapy (e.g., within about any of one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to the tyrosine kinase inhibitor and / or the electrostatic therapy).In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the electrostatic treatment. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the electrostatic treatment. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered sequentially (e.g., prior to or after the tyrosine kinase inhibitor and / or the electrostatic treatment). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered immediately after the tyrosine kinase inhibitor and / or the electrostatic treatment (e.g., within any of about 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours after the tyrosine kinase inhibitor and / or the electrostatic treatment).

[0102] In some embodiments, two or more proinflammatory agents described herein are administered to an individual. For example, in some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering a tyrosine kinase inhibitor, a TLR agonist, or a STING activator (e.g., MSA-2, ADU-S100, or cGAMP) and an immune checkpoint inhibitor. In some embodiments, the TLR agonist activates one or more TLRs selected from the group consisting of TLR9, TLR4, TLR7, and TLR8. In some embodiments, the immune checkpoint inhibitor is an anti-PD-1 agent (such as an anti-PD-1 antibody), an anti-PD-L1 agent (such as an anti-PD-L1 antibody), or an anti-CTLA-4 agent (such as an anti-CTLA-4 antibody). In some embodiments, the tyrosine kinase inhibitor, the TLR agonist, and the immune checkpoint inhibitor are administered within the same day. In some embodiments, the tyrosine kinase inhibitor is administered intermittently. In some embodiments, the tyrosine kinase inhibitor, the TLR agonist, and / or the immune checkpoint inhibitor is administered at least twice (e.g., at least three times, four times, five times, or six times). In some embodiments, the tyrosine kinase inhibitor, the TLR agonist, and the immune checkpoint inhibitor are administered for at least two cycles (e.g., at least three cycles), optionally with at least two consecutive days (e.g., at least three consecutive days) within the same day in each cycle. In some embodiments, each cycle has about seven days to about twenty days. In some embodiments, the TLR agonist activates a TLR on a macrophage, optionally wherein the TLR comprises TLR9. In some embodiments, the TLR agonist activates at least two TLRs (e.g., TLR4, TLR7, TLR8, or TLR9). In some embodiments, the TLR agonist activates at least three TLRs (e.g., TLR9, TLR4, and TLR7 / 8). In some embodiments, the TLR agonist comprises CpG, poly I:C, and / or R848. In some embodiments, the TLR agonist is selected from the group consisting of LTA, CpG, Poly I;C, LPS, MPLA, flagellin, R848, vesatolimod, bromopindol, motolimod, and losoxantrone. In some embodiments, the TLR agonist comprises CpG, poly I:C, and R848, e.g., at a ratio of 1 : 1 : 1. In some embodiments, the tyrosine kinase inhibitor is administered systemically, and the TLR agonist is administered intratumorally. In some embodiments, the tyrosine kinase inhibitor is administered systemically and intratumorally. In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling. In some embodiments, the tyrosine kinase inhibitor inhibits Src family kinases (SFKs). In some embodiments, the tyrosine kinase inhibitor is a Hck inhibitor.In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406. In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof). In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered prior to (e.g., within any of about one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to) the tyrosine kinase inhibitor and / or the TLR agonist and / or the immune checkpoint inhibitor. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the TLR agonist and / or the immune checkpoint inhibitor. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the TLR agonist and / or the immune checkpoint inhibitor. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered sequentially (e.g., prior to or after) the tyrosine kinase inhibitor and / or the TLR agonist and / or the immune checkpoint inhibitor. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered immediately after (e.g., within any of about 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours, after) the tyrosine kinase inhibitor and / or the TLR agonist and / or the immune checkpoint inhibitor. In some embodiments, the TLR agonist is selected from the group consisting of LTA, CpG, Poly I;C, LPS, MPLA, flagellin, R848, vesatolimod, bromopindol, motolimod, and losoxantrone.

[0103] In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., a cancer that is resistant or refractory to a checkpoint inhibitor, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor (e.g., a Src family kinase inhibitor, such as any of the inhibitors listed in Table 2), a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof), and a TLR agonist or STING activator as discussed herein. In some embodiments, the TLR agonist is selected from the group consisting of LTA, CpG, Poly I;C, LPS, MPLA, flagellin, R848, resiquimod, bropirimine, motolimod, and loxoribine. In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody).

[0104] In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor, wherein the individual is selected for treatment based on having an ongoing inflammatory response. In some embodiments, the individual has an acute inflammatory response. In some embodiments, the inflammatory response is in the tumor. In some embodiments, the inflammatory response is at a site different from the tumor. In some embodiments, the individual has an inflammatory response when there are at least two (e.g., two, three, four, or five) events selected from the group consisting of: a) an increase in one or more (e.g., at least one, two, three, four, five) inflammatory cytokines (such as IFNy, IL-12b, TNFa, IL-6, IL-1b, IFN-a1, IFN-a2, IFN-b1), b) a decrease in one or more (e.g., at least one, two, or three) anti-inflammatory cytokines (such as TGFb1, TGFb2, TGFb3), c) an increase in infiltrating immune cells (such as T cells, NK cells, macrophages, neutrophils), d) a decrease in suppressive immune cells (such as MDSCs), and / or e) an increase in one or more (e.g., at least one, two, three, four, or five) immunostimulatory costimulatory molecules (such as CD80, CD86, OX40L, CD40, ICOS-L, PD-L1, GITRL) in a tissue (e.g., tumor tissue) or immune cells (such as macrophages). In some embodiments, the tyrosine kinase inhibitor is administered intermittently. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of a small molecule, a nucleic acid (e.g., siRNA, shRNA, antisense RNA, microRNA), a nucleic acid editing system (e.g., a CRISPR system), and a protein agent (e.g., an antibody agent that targets a tyrosine kinase or an activated tyrosine kinase). In some embodiments, the tyrosine kinase inhibitor is administered at least two times (e.g., at least three, four, five, or six times). In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual at least two times at an interval of no more than once every three days. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual for at least two cycles, wherein the tyrosine kinase inhibitor is administered at least once in each cycle, and wherein each cycle has about three days to about twenty days. In some embodiments, the tyrosine kinase inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or locally (e.g., intratumorally). In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling. In some embodiments, the tyrosine kinase inhibitor inhibits Src family kinases (SFKs). In some embodiments, the tyrosine kinase inhibitor is an Hck inhibitor.In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406. In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof). In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered prior to (e.g., within any of about one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to) the tyrosine kinase inhibitor. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered sequentially (e.g., prior to or subsequent to) the tyrosine kinase inhibitor. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered immediately after (e.g., within any of about 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours, after) the tyrosine kinase inhibitor.

[0105] In some embodiments, a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual is provided, comprising administering to the individual a tyrosine kinase inhibitor, wherein the individual is selected for treatment based on having ongoing immunogenic cell death (ICD). In some embodiments, the individual has ICD when a sample from the cancer has a higher level of one or more (e.g., at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% higher) DAMP than a reference sample (e.g., a corresponding sample in a healthy control, e.g., a sample from the cancer prior to administration of an ICD-inducing treatment). In some embodiments, the tyrosine kinase inhibitor is administered intermittently. In some embodiments, the DAMP is selected from the group consisting of endoplasmic reticulum (ER) chaperone molecules (e.g., calreticulin (CALR), e.g., heat shock proteins (HSPs)), non-histone chromatin-binding protein high mobility group box 1 (HMGB1), cytoplasmic protein annexin A1 (ANXA1), and small metabolites ATP and type I interferon (IFN). In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of small molecules, nucleic acids (e.g., siRNA, shRNA, antisense RNA, microRNA), nucleic acid editing systems (e.g., CRISPR systems), and protein agents (e.g., antibody agents targeting tyrosine kinases or activated tyrosine kinases). In some embodiments, the tyrosine kinase inhibitor is administered at least two times (e.g., at least three, four, five, or six times). In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual at least two times at intervals of no more than once every three days. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual for at least two cycles, wherein the tyrosine kinase inhibitor is administered at least once in each cycle, and wherein each cycle has about three days to about twenty days. In some embodiments, the tyrosine kinase inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or locally (e.g., intratumorally). In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling. In some embodiments, the tyrosine kinase inhibitor inhibits Src family kinases (SFKs). In some embodiments, the tyrosine kinase inhibitor is an Hck inhibitor. In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406. In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof).In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody). In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered prior to (e.g., within about any of one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to) the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered sequentially (e.g., prior to or after) the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered immediately after (e.g., within about any of 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours, after) the tyrosine kinase inhibitor and / or the proinflammatory agent.

[0106] In some embodiments, the application provides a method of treating a cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an end-stage cancer) in an individual, comprising administering to the individual a) a monocyte or macrophage deficient in expression or activation of a tyrosine kinase and b) a proinflammatory agent (e.g., a TLR agonist, e.g., radiation therapy). In some embodiments, the monocyte or macrophage is derived from the same individual. In some embodiments, the monocyte or macrophage is engineered to express a chimeric receptor targeting a tumor antigen. In some embodiments, the monocyte or macrophage and the proinflammatory agent are administered within 24 hours of each other (e.g., within 12 hours, 8 hours, 4 hours, 2 hours, or 1 hour of each other, or within 30 minutes of each other). In some embodiments, the monocyte or macrophage and the proinflammatory agent are administered simultaneously, concurrently, or sequentially. In some embodiments, the monocyte or macrophage is administered prior to the proinflammatory agent. In some embodiments, the monocyte or macrophage is administered after the proinflammatory agent. In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof). In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered prior to the monocyte or macrophage and / or the proinflammatory agent (e.g., within about any of one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to the monocyte or macrophage and / or the proinflammatory agent). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered simultaneously with the monocyte or macrophage and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the monocyte or macrophage and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered sequentially with the monocyte or macrophage and / or the proinflammatory agent (e.g., prior to or after the monocyte or macrophage and / or the proinflammatory agent).In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered immediately after (e.g., within about any of 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours after) the monocyte or macrophage and / or the proinflammatory agent. In some embodiments, the TLR agonist is selected from the group consisting of LTA, CpG, Poly I;C, LPS, MPLA, flagellin, R848, resiquimod, bropirimine, motolimod, and losoxantrone.

[0107] The application also provides a method of modulating monocytes or macrophages derived from an individual having cancer, comprising contacting the monocytes or macrophages with a tyrosine kinase inhibitor as described above and a proinflammatory agent as described above. In some embodiments, the monocytes or macrophages are derived from the same individual. In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof). In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody). In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered prior to (e.g., within about any of one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to) the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered sequentially (e.g., prior to or after) the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered immediately after (e.g., within about any of 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours, after) the tyrosine kinase inhibitor and / or the proinflammatory agent.

[0108] The application also provides a method of activating phagocytosis of tumor cells in an individual having a tumor, comprising administering to the individual a tyrosine kinase inhibitor, wherein the individual a) has received, is receiving, or will receive a proinflammatory agent, or b) is in an inflammatory response or has an ongoing infection. In some embodiments, the tyrosine kinase inhibitor is administered systemically (e.g., intravenously or subcutaneously). The application also provides a method of activating tumor infiltrating T cells in an individual having a tumor, comprising administering to the individual a tyrosine kinase inhibitor, wherein the individual a) has received, is receiving, or will receive a proinflammatory agent, or b) is in an inflammatory response or has an ongoing infection. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual at least twice at intervals of no more than once every three days. In some embodiments, the method comprises administering the tyrosine kinase inhibitor to the individual for at least two cycles, wherein the tyrosine kinase inhibitor is administered at least once in each cycle, and wherein each cycle has about three days to about twenty days. In some embodiments, the proinflammatory agent and the tyrosine kinase inhibitor are administered within 24 hours of each other. In some embodiments, the proinflammatory agent comprises an agent selected from the group consisting of a TLR agonist, a STING activator, a radiation therapy, a PAMP / DAMP activator, a checkpoint inhibitor, a proinflammatory cytokine, a chemotherapeutic agent, a bacterial component, a cancer vaccine, and an oncolytic virus. In some embodiments, the method further comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof). In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFa antibody or an anti-IL-6 antibody). In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered prior to (e.g., within any of about one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to) the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces an inflammatory cytokine cascade or cytokine storm (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered concurrently with the tyrosine kinase inhibitor and / or the proinflammatory agent.In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered sequentially (e.g., prior to or after the tyrosine kinase inhibitor and / or proinflammatory agent) with the tyrosine kinase inhibitor and / or proinflammatory agent. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., a TNFα inhibitor, e.g., an anti-TNFα antibody) is administered immediately after (e.g., within any of about 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours after) the tyrosine kinase inhibitor and / or proinflammatory agent. In some embodiments, the TLR agonist is selected from the group consisting of LTA, CpG, Poly I;C, LPS, MPLA, flagellin, R848, vesatolimod, bromopindol, motolimod, and losoxantrone.

[0109] The above-described methods of cancer treatment can also be used to 1) activate the SHP-1 signaling pathway in the individual; 2) deplete the tyrosine kinase-iR-SHP-1 axis immunosuppression in the individual; 3) activate the anti-cancer innate and / or adaptive immunity within the tumor in the individual; 4) release the TLR-induced proinflammatory response; and 5) increase antigen presentation by tumor-associated macrophages (TAMs). Accordingly, the present application also provides methods for any one or more of these purposes.

[0110] Tumor microenvironment (TME) immunosuppression and SHP-1 signaling

[0111] Src Homology 2 (SH-2) domain-containing phosphatase 1 (SHP-1) is a non-receptor tyrosine phosphatase encoded by the PTPN6 gene located on human chromosome 12p13 and containing two promoter regions within exons 1 and 2, yielding two forms of SHP-1 that differ in their N-terminal amino acid sequence but have similar phosphatase activity. Promoter I is active in non-hematopoietic cells, while promoter II is active in hematopoietic-derived cells; in some epithelial cancer cells, both promoters can be functional and yield various SHP-1 alternative transcripts. The two SHP-1 isoforms show different subcellular localization: type I is mainly located in the nucleus, while type II is located in the cytoplasm, suggesting that they have different targets.

[0112] SHP-1 is a 595 amino acid protein composed of two N-terminal SH2 domains (N-SH2 and C-SH2) in tandem, a classical catalytic protein tyrosine phosphatase (PTP) domain, and a C-terminal tail containing several phosphorylation sites. Its crystal structure reveals a configuration in which N-SH2 binds to the catalytic site of the protein through charge-charge interactions. In this autoinhibited inactive state, substrate access to the active site is prevented, but binding of a phosphotyrosine residue to the SH2 domain induces a conformational change that weakens the interaction between N-SH2 and the catalytic domain. This opens the conformation to allow substrate access and is further stabilized by a new interaction between the SH2 domain and the catalytic domain. These molecular rearrangements determine a complex regulatory mechanism controlled by substrate recruitment.

[0113] Another activation mechanism is mediated by phosphorylation of amino acids within the C-terminal tail. Three phosphorylation sites have been found so far, two tyrosine (Tyr536 and Tyr564) and one serine (Ser591) residues. Tyr536 and Tyr564 are phosphorylated upon various stimuli (i.e. insulin stimulation or inducers of apoptosis), leading to an increase in SHP-1 activity. The molecular mechanism is not well understood, although it has been proposed that Tyr phosphorylation can lead to an interaction with the N-SH2 domain, thereby releasing the inhibition of the protein tyrosine phosphatase activity by this domain. SHP-1 activity can also be negatively regulated by phosphorylation of Ser591 by protein kinase C (PKC) or mitogen-activated protein kinase (MAPK), the inhibitory mechanism of which has not been well characterized.

[0114] Protein-tyrosine phosphorylation is a reversible post-translational modification that is tightly regulated by both kinases and phosphatases. Any deviation from the phosphorylation / dephosphorylation balance can promote the intracellular accumulation of tyrosine-phosphorylated proteins, which leads to altered regulation of cellular processes including cell growth, migration, invasion, differentiation, survival and cell trafficking. In this context, SHP-1 acts as a classical tumor suppressor involved in the homeostatic maintenance of potentially all these processes. SHP-1 function is indeed altered in solid and hematological human cancers by somatic mutations or epigenetic mechanisms. In addition to its well-documented role in the regulation of hematopoietic cell biology, SHP-1 has now been associated with many signal transduction pathways relevant to cancer pathogenesis and progression.

[0115] However, inhibition of SHP-1 is at risk of having serious side effects. Mice deficient in the SHP-1 gene, i.e. moth-eaten mice (me / me or me v / me vStudies in SHP-1 deficient mice have revealed severe immune abnormalities and immune cell hyperactivation associated with the global loss of tyrosine kinase phosphatases. Wormy mice usually die from life-threatening autoimmune inflammatory disorders in early childhood. Even partial depletion of SHP-1 in wild-type mice after adulthood leads to features of inflammatory disease, causing extensive lung inflammation and splenomegaly. Inhibition of SHP-1 is a double-edged sword, with the potential to enhance anti-cancer immunity but inevitably jeopardizing the host's enhanced inflammatory responses, cytokine storm, and autoimmunity.

[0116] Inhibitors targeting the activity of tyrosine kinase phosphatases have been developed for some time, and some of them have now entered preclinical studies, including NSC-87877, stannsoporfin (SSG), tyrosine phosphatase inhibitor 1 (TPI-1 or analogs or derivatives thereof), and suramin; however, only a few of them have shown activity in experimental tumor models. SSG has been tested in phase I trials for both malignant melanoma (NCT00498979) and advanced malignancies (NCT00629200); the drug was administered in combination with interferon, followed or not by chemotherapy treatment. Unfortunately, no effect on tumor progression was observed, and the most common toxic side effects were thrombocytopenia, elevated serum lipases, fatigue, fever, chills, anemia, hypokalemia, pancreatitis, and skin rash (observed in up to 68% of patients). Currently, no SHP-1 inhibitor is in phase II trials.

[0117] The SHP-1 inhibitors described herein can be administered with a tyrosine kinase inhibitor. In some embodiments, the method comprises administering to the individual (e.g., locally or systemically) an effective amount of a SHP-1 inhibitor (e.g., TPI-1 or analogs or derivatives thereof). In some embodiments, the SHP-1 is administered simultaneously with the tyrosine kinase inhibitor. In some embodiments, the SHP-1 is administered sequentially (e.g., before or after the tyrosine kinase inhibitor) with the tyrosine kinase inhibitor. In some embodiments, the SHP-1 administration follows the same dosing regimen as the tyrosine kinase inhibitor.

[0118] Agents that reduce systemic inflammation

[0119] In some cases, individuals develop systemic inflammation, i.e., cytokine release syndrome (CRS), after receiving, for example, immunotherapeutic treatments, however the inflammatory condition is not fully understood. CRS can be induced by direct target cell lysis and sequential release of cytokines such as TNFa or IFNy, or by T cell activation followed by cytokine release due to treatment stimulation. These cytokines trigger a chain reaction due to activation of innate immune cells such as macrophages and endothelial cells, which then induce further cytokine release. In particular, IL-6, IL-10, and IFNy are most commonly found elevated in patients with CRS.

[0120] The methods described herein can also include administration of an agent that reduces systemic inflammation (including, for example, an agent that reduces the inflammatory cytokine cascade or cytokine storm, e.g., a TNFα inhibitor such as an anti-TNFα antibody) to suppress systemic inflammation and reduce adverse toxicities. These agents include, but are not limited to, inhibitors of TNFα, IL-6, IL-10, and IFNγ. In some embodiments, the agent that reduces systemic inflammation (e.g., a TNFα inhibitor such as an anti-TNFα antibody) is administered prior to (e.g., within any of about one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day, or less, prior to) the tyrosine kinase inhibitor. In some embodiments, the agent that reduces systemic inflammation (e.g., a TNFα inhibitor such as an anti-TNFα antibody) is administered concurrently with the tyrosine kinase inhibitor. In some embodiments, the agent that reduces systemic inflammation (e.g., a TNFα inhibitor such as an anti-TNFα antibody) is administered concurrently with the tyrosine kinase inhibitor. In some embodiments, the agent that reduces systemic inflammation (e.g., a TNFα inhibitor such as an anti-TNFα antibody) is administered sequentially (e.g., prior to or after) the tyrosine kinase inhibitor. In some embodiments, the agent that reduces systemic inflammation (e.g., a TNFα inhibitor such as an anti-TNFα antibody) is administered immediately after (e.g., within any of about 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours, after) the tyrosine kinase inhibitor. In some embodiments, the administration of the agent that reduces systemic inflammation (e.g., a TNFα inhibitor such as an anti-TNFα antibody) follows the same dosing regimen as the tyrosine kinase inhibitor. In some embodiments, the agent that reduces systemic inflammation (e.g., a TNFα inhibitor such as an anti-TNFα antibody) is administered at a subtherapeutic dose, i.e., at a dose lower than the effective amount for treating the disease when administered alone. In some embodiments, the administration of the agent that reduces systemic inflammation (e.g., a TNFα inhibitor such as an anti-TNFα antibody) allows for more frequent administration of the tyrosine kinase inhibitor and / or pro-inflammatory agent (e.g., daily, every other day, every third day, etc.).

[0121] The agent can include any anti-inflammatory agent known in the art, including inhibitors or antagonists of pro-inflammatory agents. For example, the agent can be an inhibitor or antagonist, including but not limited to small molecule inhibitors, neutralizing antibodies, receptor blocking antibodies, soluble receptors, targeted short interfering RNA (siRNA), chemical inhibitors of mRNA stability, analogs or derivatives thereof, and any combination thereof, including combinations of agents that target one or more molecules (e.g., targeting via inhibition of TNFα alone, inhibition of IL-6 alone, or inhibition of both TNFα and IL-6 in combination).

[0122] Anti-TNFa antagonists

[0123] The primary proinflammatory cytokine, TNFa, is secreted by activated macrophages, monocytes, and lymphocytes. The inventors have surprisingly found that administration of an anti-TNFa antibody to an individual who has been administered a tyrosine kinase inhibitor and a proinflammatory agent reduces toxicity caused by systemic inflammation without compromising the efficacy of the therapeutic agent.

[0124] Accordingly, in some embodiments, the methods of the present application comprise administration of a TNFa inhibitor, e.g., an anti-TNFa antagonist (e.g., where the proinflammatory agent is not TNFa). In some embodiments, the TNFa inhibitor is selected from the group consisting of small molecule inhibitors, neutralizing antibodies, TNFa receptor blocking antibodies, soluble TNFa receptors, TNFa targeting short interfering RNA (siRNA), chemical inhibitors of TNFa mRNA stability, inhibitors of TNFa converting enzyme (TACE), and analogs or derivatives thereof. In some embodiments, the TNFa inhibitor is an anti-TNFa neutralizing antibody. In some embodiments, the TNFa inhibitor is an anti-TNFa receptor blocking antibody. In some embodiments, the anti-TNFa antibody is a monoclonal antibody. In some embodiments, the anti-TNFa antibody is a chimeric, humanized, and / or fully human antibody.

[0125] Suitable antibodies for use in the methods provided herein include, but are not limited to Remicade® (infliximab (Centocor)), and, for example, those described in U.S. Patent Nos. 6,835,823; 6,790,444; 6,284,471; 6,277,969; 5,919,452; 5,698,195; 5,656,272; and 5,223,395; and European Patent No. 0610201, or antibodies that bind to the same epitope as Suitable anti-TNFa antibodies for use in the methods provided herein are, by way of non-limiting example, Humira® (adalimumab (Abbott Laboratories, Esa i)), as described in U.S. Patent Nos. 6,090,382; 6,258,562; or 6,509,015, and related patents and applications, the contents of each of which are hereby incorporated by reference in their entirety; Simponi® (golimumab (Janssen Biotech, Inc.)), as described in PCT Publication No. WO 02 / 12502, and related patents and applications; Cimzia® (certolizumab pegol (UCB, Inc.)), as described in U.S. Patent No. 6,905,960, and related patents and applications; and Yervoy® (ipilimumab (Bristol-Myers Squibb)), as described in U.S. Patent No. 7,208, 159, and related patents and applications. TM(Cimzia, CDP 571 (Biogen Idee), Raptiva (Synovir), Humira (adalimumab, Abbott), Simponi (golimumab, CNTO 148 (Centocor)), the contents of these documents are hereby incorporated by reference in their entirety; ART621 (Arana Therapeutics), SSS 07 (Epitopmics and 3SBio), or antibodies that bind to the same epitope as Humira, Simponi, ART621, or SSS 07.

[0126] In some embodiments, the TNFa inhibitor, e.g., an anti-TNFa antagonist, is a fusion protein. Suitable fusion proteins for use in the methods provided herein include, but are not limited to, Enbrel (etanercept (Amgen)) and other fusion proteins or fragments thereof described in U.S. Patent No. 5,712,155, PCT Publication No. WO 91 / 03553, and related patents and applications, the contents of which are hereby incorporated by reference in their entirety.

[0127] In some embodiments, the TNFa inhibitor, e.g., an anti-TNFa antagonist, is a modified antibody antagonist or a non-antibody-based antagonist. Such antagonists include advanced antibody therapeutics, such as antibody fragments, including but not limited to Cimzia TM (Cimzia, CDP 571 (Biogen Idee), Raptiva (Synovir), Humira (adalimumab, Abbott), Simponi (golimumab, CNTO 148 (Centocor)), the contents of these documents are hereby incorporated by reference in their entirety; ART621 (Arana Therapeutics), SSS 07 (Epitopmics and 3SBio), or antibodies that bind to the same epitope as Humira, Simponi, ART621, or SSS 07. such as ABX 0402 (Ablynx), immunotoxins and radiolabeled therapeutic agents; peptide therapeutics; gene therapy, particularly intrabodies; oligonucleotide therapeutics, such as aptamer therapeutics, antisense therapeutics, interfering RNA therapeutics; and small molecules, such as LMP-420 (LeukoMed) as described in European Patent No. 0767793, and related patents and applications, the contents of which are hereby incorporated by reference in their entirety.

[0128] In some embodiments, the TNFa inhibitor is administered systemically. In some embodiments, the TNFa inhibitor is administered at least once a week, once every five days, once every three days, or daily. In some embodiments, the TNFa inhibitor is administered intermittently. In some embodiments, the individual is administered the TNFa inhibitor for at least two cycles, wherein each cycle has from about three days to about seven days. In some embodiments, the individual does not develop cytokine release syndrome or proinflammatory organ damage. In some embodiments, the administration of the TNFa inhibitor does not impair or slightly impairs tumor clearance.

[0129] In some embodiments, the TNFa inhibitor is administered prior to (e.g., within any of about two weeks, one week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, 1 day or less, prior to) the tyrosine kinase inhibitor and / or proinflammatory agent. Exemplary TNFa inhibitors such as anti-TNFa antibodies are typically stable for at least one or two weeks. In some embodiments, the TNFa inhibitor is administered concurrently with the tyrosine kinase inhibitor and / or proinflammatory agent. In some embodiments, the TNFa inhibitor is administered concurrently with the tyrosine kinase inhibitor and / or proinflammatory agent. In some embodiments, the TNFa inhibitor is administered sequentially with the tyrosine kinase inhibitor and / or proinflammatory agent (e.g., prior to or after the tyrosine kinase inhibitor and / or proinflammatory agent). In some embodiments, the TNFa inhibitor is administered immediately after (e.g., within any of about 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 1 hour, 2 hours, or 3 hours, after) the tyrosine kinase inhibitor and / or proinflammatory agent.

[0130] Anti-IL6 antagonists

[0131] An“anti-IL6 antagonist” or“IL6 inhibitor” refers to an agent that inhibits or blocks IL6 biological activity via binding to IL6 or IL6 receptor. In some embodiments, the anti-IL6 antagonist is an antibody. In one embodiment, the anti-IL6 antagonist is an antibody that binds IL6 receptor. Antibodies that bind IL-6 receptor include tocilizumab (including its intravenous (i.v.) and subcutaneous (s.c.) formulations) (Chugai, Roche, Genentech), satralizumab (Chugai, Roche, Genentech), sarilumab (Sanofi, Regeneron), NI-1201 (Novimmune and Tiziana), and vobarilizumab (Ablynx). In one embodiment, the anti-IL6 antagonist is a monoclonal antibody that binds IL6. Antibodies that bind IL-6 include sirukumab (Centecor, Janssen), olokizumab (UCB), clazakizumab (BMS and Alder), siltuximab (Janssen), EBI-031 (Eleven Biotherapeutics and Roche). In one embodiment, the IL6 antagonist is olokizumab.

[0132] In some embodiments, the IL6 inhibitor is administered systemically. In some embodiments, the IL6 inhibitor is administered at least once a week, once every five days, once every three days, or once a day. In some embodiments, the IL6 inhibitor is administered intermittently. In some embodiments, the IL6 inhibitor is administered to the individual for at least two cycles, wherein each cycle has from about three days to about seven days.

[0133] Tyrosine kinase inhibitor

[0134] As referred to herein, a tyrosine kinase inhibitor is any kind or class of agent that inhibits the expression or activation of a tyrosine kinase.

[0135] In some embodiments, the tyrosine kinase inhibitor is capable of inhibiting at least about 20% (e.g., at least 20%, 30%, 40%, or 50%) of tyrosine kinase activity. In some embodiments, the tyrosine kinase inhibitor is capable of inhibiting at least about 20% (e.g., at least 20%, 30%, 40%, or 50%) of tyrosine kinase expression.

[0136] In some embodiments, the tyrosine kinase inhibitor specifically inhibits SHP-1 signaling.

[0137] In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of a small molecule, a nucleic acid (e.g., siRNA, shRNA, antisense RNA, microRNA), a nucleic acid editing system (e.g., a CRISPR system), a protein agent (e.g., an antibody agent that targets a tyrosine kinase or an activated tyrosine kinase, e.g., a dominant negative tyrosine kinase or a constitutively active tyrosine kinase mutant).

[0138] In some embodiments, the tyrosine kinase inhibitor has a half-life of no more than about 10 days, 9 days, 8 days, or 7 days (e.g., a half-life of no more than about 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or 1 day).

[0139] In some embodiments, the tyrosine kinase inhibitor is effective to inhibit more than 50% of tyrosine kinase activity for no more than about 10 days, 9 days, 8 days, 7 days, 6 days, or 5 days. In some embodiments, the tyrosine kinase inhibitor is effective to inhibit more than 50% of tyrosine kinase activity for no more than 4 days, 3 days, 2 days, or 1 day.

[0140] In some embodiments, the tyrosine kinase inhibitor is a covalent inhibitor. In some embodiments, the tyrosine kinase inhibitor is a non-covalent inhibitor.

[0141] In some embodiments, the tyrosine kinase inhibitor is a competitive inhibitor.

[0142] In some embodiments, the tyrosine kinase inhibitor is a nucleic acid editing system, such as a CRISPR system. In some embodiments, the CRISPR components are introduced into the cell (e.g., monocytes and macrophages), but the DNA encoding the guide RNA or Cas9 is not incorporated into the genome of the cell. Under this approach, the CRISPR system only cuts the genomic DNA of the cell for a limited period of time. See, e.g., Fister et al., Front Plant Sci. 2018 Mar 2;9:268.

[0143] In some embodiments, the tyrosine kinase inhibitor is administered at least twice (such as at least 3 times, 4 times, 5 times, or 6 times).

[0144] In some embodiments, the method comprises administering the tyrosine kinase inhibitor at least twice (such as at least three times, four times, five times, or six times) at an interval of no more than once every two days.

[0145] In some embodiments, the method comprises administering the tyrosine kinase inhibitor at least twice (such as at least three times, four times, five times, or six times) at an interval of no more than once every three days.

[0146] In some embodiments, the method comprises administering the tyrosine kinase inhibitor for at least two cycles. In some embodiments, the tyrosine kinase inhibitor is administered at least once (e.g., twice, three times, four times) in each cycle. In some embodiments, each cycle has about three days to about 50 days (e.g., about 3 days-40 days, about 3 days-30 days, about 3 days-20 days, about 3 days-15 days, about 3 days-10 days, or about 2 days-10 days).

[0147] In some embodiments, the tyrosine kinase inhibitor is administered systemically (e.g., orally, intravenously, subcutaneously, intraperitoneally). In some embodiments, the tyrosine kinase inhibitor is administered locally (e.g., intratumorally). In some embodiments, the tyrosine kinase inhibitor is administered systemically and locally (e.g., intratumorally).

[0148] In some embodiments, the tyrosine kinase inhibitor is complexed with a delivery vehicle prior to administration to the individual. In some embodiments, the delivery vehicle facilitates delivery into the tumor.

[0149] In some embodiments, the tyrosine kinase inhibitor modulates monocytes or macrophages (e.g., monocytes or macrophages derived from the individual to be treated) in vitro.

[0150] In some embodiments, the tyrosine kinase inhibitor and the proinflammatory agent below are administered within 24 hours of each other (e.g., within 12 hours, 8 hours, 4 hours, 2 hours, or 1 hour of each other, or within 30 minutes of each other). In some embodiments, the tyrosine kinase inhibitor and the proinflammatory agent are administered simultaneously, concurrently, or sequentially. In some embodiments, the tyrosine kinase inhibitor is administered prior to the proinflammatory agent. In some embodiments, the tyrosine kinase inhibitor is administered after the proinflammatory agent.

[0151] Phosphorylation of Src family tyrosine kinases (SFKs) and ITIMs in TAMs

[0152] In some embodiments, the tyrosine kinase is a tyrosine kinase of the Src family. Src family kinases have a similar structure consisting of an N-terminal Src homology (“SH”) 4 (“SH4”) domain, a “unique” domain, an SH3 domain, an SH2 domain, a catalytic domain (also referred to as SH1 domain or kinase domain), and a short C-terminal tail. Activity is regulated by tyrosine phosphorylation at two sites. Phosphorylation of a tyrosine in the C-terminal tail (Tyr-505, Src numbering) leads to down-regulation by promoting intramolecular interactions between the tail and the SH2 domain. The eight known mammalian members of the Src family are divided into two subfamilies. Lck is most similar to Hck, Lyn, and Blk (more than 65% identity between any two members). The other subfamily consists of Src, Yes, Fyn, and Fgr (more than 70% identity between any two members). Residues important for Src family kinase activity and / or substrate specificity have been identified by X-ray crystal structures and by structure modeling studies, and are highly conserved among family members.

[0153] The non-receptor tyrosine kinase Src family (SFKs) consists of SRC, LCK, LYN, BLK, HCK, FYN, FGR, and YES (8 / 9 members are expressed in humans) and can be divided into two groups according to their expression patterns. SRC, YES, and FYN are ubiquitously expressed, while LCK, FGR, BLK, LYN, YRK, and HCK show specific expression in certain types of cells and tissues.

[0154] In the immune system, SFKs play important regulatory functions in myeloid and lymphoid lineage immune cells, controlling cell activation, proliferation, differentiation, apoptosis, cytokine production, migration, metabolism, etc.

[0155] LCK is specifically expressed in T cells and is critically involved in TCR-mediated T cell activation; lack of LCK renders TCR signaling ineffective, thus reducing antigen-specific T cell activation, proliferation, and T cell immunity. LCK is not expressed in macrophages or other myeloid leukocytes.

[0156] LYN is highly expressed in B cells and also in myeloid leukocytes. In macrophages, our studies found that LYN remains constitutively active and mediates low level tyrosine phosphorylation in the cytoplasmic ITIM of iRs (inhibitory receptors). However, LYN does not appear to be involved in the strong ITIM tyrosine phosphorylation of iRs stimulated by stimuli. In particular, we found that under tumor therapy conditions, HCK or its related complementary SFKs (e.g. FGR and YES; see: Lowell CA, Soriano P, Varmus HE. Functional overlap in the src gene family: inactivation of hck and fgr impairs natural immunity. Genes and Development. 1994; 8: 387-398.) phosphorylate the ITIM of iRs, resulting in docking and activation of SHP-1, which mediates downstream inhibitory regulation.

[0157] Figure 12 Examples are shown by studying SIRPa, an iR that is abundantly expressed in tumor-associated macrophages (TAMs). As shown, pro-inflammatory stimuli (TLR agonists, pro-inflammatory cytokines IL-1 b, IL-6, IL-12, IL-17, IL-18, TNFa, IFNy, etc. and cancer therapy) induce SIRPa ITIM phosphorylation and exclusive binding of SHP-1 (not SHP-2). Anti-inflammatory cytokine stimuli also induce SIRPa ITIM phosphorylation, but bind to SHP-2.

[0158] In the case of macrophage activation by pro-inflammatory stimuli or cancer therapy, inhibition of Src family tyrosine kinases (SFKs) by PP1 and PP2 (both SFK inhibitors), but not by inhibitors targeting other TKs such as JAK (JAK inh.), Btk (LFMA-13) or Syk (paclitaxel), reduces SIRPa cytoplasmic ITIM phosphorylation and binding of SIRPa to SHP-1. In contrast, the specific inhibitor of LYN, bafetinib (also known as INNO-406), has only a small effect. Although in the absence of pro-inflammatory stimuli, Lyn lacks a significant impact on the low level SIRPa ITIM phosphorylation induced by CD47 ligation, the tested macrophages lacking Lyn confirm that Lyn has no impact on pro-inflammatory factor-induced SIRPa ITIM phosphorylation and binding of SIRPa to SHP-1.

[0159] Notably, in the absence of therapy, the immunosuppressive tumor TME is controlled by IL-10, TGF and IL-4 / 13, which activate Bruton's tyrosine kinase (Btk) in macrophages (TAMs), leading to cytoplasmic ITIM phosphorylation of iR (e.g., SIRPa) and docking of SHP-2, but not SHP-1. This axis of events, in which immunosuppressive cytokines activate Btk to drive SIRPa-SHP-2 binding, further enhances immunosuppressive signaling within TAMs. As another consequence of this pathway, iR expression on TAMs is further increased, serving as a feed-forward mechanism to control TAMs, and thus, TME immunosuppression.

[0160] In some embodiments, the TKi inhibits a Src family kinase (SFK), optionally wherein the SFK is selected from the group consisting of SRC, LCK, LYN, BLK, HCK, FYN, FGR, and YES. In some embodiments, the TKi inhibits a SFK that is not LYN. In some embodiments, the TKi inhibits a SFK that is not HCK. In some embodiments, the SFK is selected from the group consisting of SRC, BLK, HCK, FYN, FGR, and YES. In some embodiments, the SFK is HCK or its related, complementary SFKs (e.g., FGR and YES). In some embodiments, the SFK is selected from the group consisting of HCK, FGR, and YES.

[0161] In some embodiments, the tyrosine kinase inhibitor is a Src inhibitor. In some embodiments, the tyrosine kinase inhibitor is a Syk inhibitor. In some embodiments, the tyrosine kinase inhibitor is a Hck inhibitor. In some embodiments, the tyrosine kinase inhibitor inhibits any one or more of (such as any one of 2, 3, 4, 5, or 6 of) Src, Syk, Hck, Lck, Lyn, and Yes. In some embodiments, the tyrosine kinase inhibitor inhibits Bcr-Abl. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406. In some embodiments, the tyrosine kinase inhibitor is selected from the group consisting of ponatinib, bosutinib, secaitinib, and KX2-391. These tyrosine kinase inhibitors are discussed further below.

[0162] Src inhibitors

[0163] Src is a member of non-receptor protein tyrosine kinases and has an activity of phosphorylating a specific tyrosine residue in a target protein. Src can be derived from any animal species (e.g., mammal), and can be, for example, at least one selected from the group consisting of primate Src, including human Src (e.g., Accession No. NP_005408), monkey Src (e.g., Accession No. XP_002830325), and the like, and rodent Src, including mouse Src (e.g., Accession No. NP_001020566), rat Src (e.g., Accession No. NP_114183), and the like, but is not limited thereto.

[0164] In some embodiments, the Src inhibitor (SRCi) can be an inhibitor of Src gene or Src protein expression; or an inhibitor of Src protein activity. The Src gene or Src protein expression inhibitor can be one or more selected from the group consisting of an antisense nucleotide, a short interfering RNA (siRNA), a short hairpin RNA (shRNA), and a ribozyme that complementarily binds to mRNA of the gene, but is not limited thereto. In addition, the Src protein activity inhibitor can be one or more selected from the group consisting of a compound, a peptide, a peptide mimetic, an aptamer, an antibody, and a natural product that specifically binds to the protein, but is not limited thereto. The antibody includes a monoclonal antibody, a polyclonal antibody, or a recombinant antibody capable of specifically binding to the Src protein, and can be constructed or purchased and used by known methods known to those skilled in the art. According to the present disclosure, the compound can be one or more selected from the group consisting of dasatinib, bosutinib, ponatinib, saracatinib, WH-4-023, KX2-391, and WZ3105.

[0165] In one embodiment, the Src inhibitor can be at least one selected from the group consisting of dasatinib, saracatinib, and bosutinib, or any combination thereof.

[0166] KX2-391 (Tibablin), also known as N-benzyl-2-(5-(4-(2-morpholinoethoxy)phenyl)pyridin-2-yl)acetamide, has the following structure:

[0167]

[0168] Dasatinib, also known as N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)-1-piperazinyl]-2-methyl-4-pyrimidinyl]amino]-5-thiazolecarboxamide monohydrate, has the following structure:

[0169]

[0170] Sunitinib, also known as SU11248, SU 12448, and ISELINIB, has the following structure:

[0171]

[0172] Bosutinib, also known as 4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4- methylpiperazin-1-yl)propoxy]quinoline-3-carbonitrile, has the following structure:

[0173]

[0174] Syk inhibitors

[0175] Spleen tyrosine kinase (Syk) is a cytosolic non-receptor protein tyrosine kinase (PTK). The human SYK gene is located in the q22 region of chromosome 9. Syk, like ZAP70, is a member of the tyrosine kinase Syk family. These cytoplasmic non-receptor tyrosine kinases share a characteristic dual SH2 domain separated by a linker domain.

[0176] In some embodiments, the Syk inhibitor can be an inhibitor of Syk gene or Syk protein expression; or an inhibitor of Syk protein activity. The Syk gene or Syk protein expression inhibitor can be one or more selected from the group consisting of an antisense nucleotide, a short interfering RNA (siRNA), a short hairpin RNA (shRNA), and a ribozyme that binds complementarily to mRNA of the gene, but is not limited thereto. In addition, the Syk protein activity inhibitor can be one or more selected from the group consisting of a compound, a peptide, a peptide mimetic, an aptamer, an antibody, and a natural product that specifically binds to the protein, but is not limited thereto. The antibody includes a monoclonal antibody, a polyclonal antibody, or a recombinant antibody that can specifically bind to the Syk protein, and can be constructed or purchased and used by known methods known to one of ordinary skill in the art.

[0177] In some embodiments, the Syk inhibitor is a small molecule inhibitor. In some embodiments, the Syk inhibitor is selected from the group consisting of entospletinib (GS-9973), fostamatinib (R788), R406, sedutinib (PRT0626070), and TAK-659.

[0178] In some embodiments, the Syk inhibitor is R406 having the following formula:

[0179]

[0180] Hck inhibitors

[0181] Hck is a member of the Src family of non-receptor tyrosine kinases that plays many roles in signaling pathways involved in the regulation of cellular processes. Hck is expressed in cells of hematopoietic origin, particularly myeloid mononuclear cells and B lymphocytes. It is involved in phagocytosis, adhesion, migration, regulation of protrusion formation on the cell membrane, lysosomal exocytosis, podosome formation, and actin polymerization. High levels of Hck are present in chronic myeloid leukemia and other hematological tumors. Hck can also play a role in the development of acute myeloid leukemia.

[0182] In some embodiments, the Hck inhibitor can be an inhibitor of Hck gene or Hck protein expression; or an inhibitor of Hck protein activity. The Hck gene or Hck protein expression inhibitor can be one or more selected from the group consisting of an antisense nucleotide, a short interfering RNA (siRNA), a short hairpin RNA (shRNA), and a ribozyme that binds complementarily to the mRNA of the gene, but is not limited thereto. In addition, the Hck protein activity inhibitor can be one or more selected from the group consisting of a compound, a peptide, a peptide mimetic, an aptamer, an antibody, and a natural product that specifically binds to the protein, but is not limited thereto. The antibody includes a monoclonal antibody, a polyclonal antibody, or a recombinant antibody that can specifically bind to the Hck protein, and can be constructed or purchased and used by known methods known to those skilled in the art.

[0183] In some embodiments, the Hck inhibitor is a small molecule inhibitor. In some embodiments, the Hck inhibitor is selected from the group consisting of RK-20449, RK-20693, RK-24466, RK-20444, RK-20445, and RK-20466. In other embodiments, the HCK inhibitor is selected from RK-20449, RK-20693, RK-24466, RK-20444, RK-20445, RK-20466, RK-20730, RK-20690, RK-20781, RK-20786, RK-20888, RK-20658, RK-20686, RK-20696, RK-20709, RK-20721, RK-20694, RK-20703, RK-20718, RK-20744, and the compounds having Hck inhibitory activity disclosed in WO2014 / 017659, which is incorporated herein by reference. Hck inhibitors are also disclosed in WO2018 / 052120, which is incorporated herein by reference.

[0184] RK-20449 (also known as A 419259): 7-((1R,4R)-4-(4-methylpiperazin-1-yl)cyclohexyl)-5-(4-phenoxyphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine has the following structure:

[0185]

[0186] Lck inhibitors

[0187] Lck (or lymphocyte-specific protein tyrosine kinase) is a member of the Src kinase family that is important for the activation of T cell receptor signaling in both naive and effector T cells. The N-terminal tail of Lck is myristoylated and palmitoylated, which tethers the protein to the plasma membrane of the cell. The protein also contains an SH3 domain, an SH2 domain, and a tyrosine kinase domain in the C-terminal portion.

[0188] In some embodiments, the Lck inhibitor can be an inhibitor of Lck gene or Lck protein expression; or an inhibitor of Lck protein activity. The Lck gene or Lck protein expression inhibitor can be one or more selected from the group consisting of an antisense nucleotide, a short interfering RNA (siRNA), a short hairpin RNA (shRNA), and a ribozyme that binds complementarily to mRNA of the gene, but is not limited thereto. In addition, the Lck protein activity inhibitor can be one or more selected from the group consisting of a compound, a peptide, a peptide mimetic, an aptamer, an antibody, and a natural product that specifically binds to the protein, but is not limited thereto. The antibody includes a monoclonal antibody, a polyclonal antibody, or a recombinant antibody that can specifically bind to the Lck protein, and can be constructed or purchased and used by known methods known to those skilled in the art.

[0189] In some embodiments, the Lck inhibitor is a small molecule inhibitor. In some embodiments, the Lck inhibitor is selected from the group consisting of Saractinib, Masitinib, and NVP-BEP800.

[0190] Bcr-Abl inhibitors

[0191] BCR-ABL (a fusion gene resulting from a reciprocal translocation mutation in the long arms of chromosomes 9 and 12) encodes the BCR-ABL protein (a constitutively active cytoplasmic tyrosine kinase present in >90% of all patients with chronic myelogenous leukemia (CML) and 15-30% of adult patients with acute lymphoblastic leukemia (ALL)). Exemplary Bcr-Abl inhibitors include, but are not limited to, imatinib, nilotinib, dasatinib, bosutinib, ponatinib, bafetinib, rebastinib, tozasertib, darinaparsen, HG-7-85-01, GNF-2, and 1,3,4-thiadiazole analogs or derivatives. Additional Bcr-Abl inhibitors can be found, for example, in WO 2006 / 052810, which is specifically incorporated by reference herein.

[0192] Ponatinib (AP24534) is a dual Src / Abl inhibitor having the following structure.

[0193]

[0194] Proinflammatory agents

[0195] Infection and tissue damage are two classic triggers of inflammation. See, e.g., Medzhitov, Nature. 2008 Jul 24;454(7203):428-35. The proinflammatory agents described herein include at least two overlapping categories: 1) any kind or class of agent or treatment that can promote inflammation (e.g., by promoting one or more proinflammatory cytokines or chemokines, inhibiting one or more anti-inflammatory cytokines or chemokines, recruiting macrophages, NK cells, neutrophils, effector T cells, or B cells to a tissue or activating any of these cells, or inhibiting regulatory / suppressive immune cells such as regulatory T cells or MDSCs), and 2) agents or treatments that can cause damage to cancer cells (e.g., cancer cell necrosis).

[0196] In some embodiments, the proinflammatory agent triggers a proinflammatory signal on macrophages. See, e.g., Figure 5A In some embodiments, the proinflammatory agent activates a TLR, TNFR, or ITAM-R. See Lionel et al., Eur J Immunol. 2011 Sep;41(9):2477-2481. The proinflammatory agent can activate a proinflammatory signal on macrophages via a direct or indirect means. For example, both a TLR agonist that directly activates a TLR on macrophages or radiation therapy that indirectly activates a proinflammatory signal on macrophages exhibit significant anti-tumor effects when used with a tyrosine kinase inhibitor. See Examples.

[0197] Exemplary pro-inflammatory agents include TLR agonists, STING activators, radiation therapy, PAMP / DAMP activators, checkpoint inhibitors, pro-inflammatory cytokines or chemokines, chemotherapy, bacterial components, cancer vaccines, and oncolytic viruses. Other exemplary pro-inflammatory agents include sound therapy (e.g., high intensity focused ultrasound) that can kill cancer cells, magnetotherapy, electrotherapy, and electrostatic therapy. See, e.g., Naud et al., Nanoscale Adv., 2020, 2, 3632-3655; Rominiyi et al., Br J Cancer. 2021 Feb; 124(4): 697-709; Zandi et al., Cancer Med. 2021 Nov; 10(21):7475-7491.

[0198] In some embodiments, the pro-inflammatory agent comprises an agent selected from the group consisting of TLR agonists, STING activators, radiation therapy, PAMP / DAMP activators, checkpoint inhibitors, pro-inflammatory cytokines or chemokines, chemotherapy, bacterial components, cancer vaccines, oncolytic viruses, sound therapy (e.g., high intensity focused ultrasound), magnetotherapy, electrotherapy, and electrostatic therapy.

[0199] In some embodiments, the pro-inflammatory agent comprises an agent selected from the group consisting of TLR agonists, STING activators, PAMP / DAMP activators, pro-inflammatory cytokines or chemokines, bacterial components, cancer vaccines, sound therapy (e.g., high intensity focused ultrasound), magnetotherapy, electrotherapy, and electrostatic therapy.

[0200] In some embodiments, the pro-inflammatory agent is sound therapy (e.g., high intensity focused ultrasound (HIFU), e.g., low intensity pulsed ultrasound (LIPUS)). See, e.g., Wood et al., Ultrasound Med Biol. 2015 Apr; 41(4):905-928; Sengupta et al., J Adv Res. 2018 Nov; 14:97-111.

[0201] In some embodiments, the pro-inflammatory agent is magnetotherapy (e.g., pulsed magnetic fields, e.g., static magnetic fields). See, e.g., Tatarov et al., Comp Med. 2011 Aug; 61(4):339-345; Sengupta et al., J Adv Res. 2018 Nov; 14:97-111.

[0202] In some embodiments, the pro-inflammatory agent is an electrotherapy or electrochemical therapy. See, e.g., Ciria et al., Chin J Cancer Res. 2013 Apr; 25(2): 223-234; Das et al., Front Bioeng Biotechnol. 2021; 9: 795300.

[0203] In some embodiments, the pro-inflammatory agent is an electrostatic therapy. See, e.g., Zandi et al., Cancer Med. 2021 Nov; 10(21): 7475-7491.

[0204] In some embodiments, the pro-inflammatory agent is a thermoacoustic therapy. See, e.g., Wen et al., Theranostics. 2017; 7(7): 1976-1989.

[0205] In some embodiments, the pro-inflammatory agent comprises or is a microorganism (e.g., a fragment or lysate of a microorganism). Examples of microorganisms include bacteria, fungi, and viruses.

[0206] TLR agonists

[0207] In some embodiments, the pro-inflammatory agent comprises or is a TLR agonist.

[0208] TLRs play a crucial role in activating immune responses. TLRs recognize conserved pathogen-associated molecular patterns (PAMPs) expressed on a variety of microorganisms, as well as endogenous DAMPs released by stressed or dying cells. TLR1, TLR2, TLR4, TLR5, TLR6, and TLR10 are expressed on the cell surface, while TLR3, TLR7, TLR8, and TLR9 are located on intracellular endosomal membranes. TLR1 and TLR2 can heterodimerize to recognize a variety of bacterial lipid structures and cell wall components, such as triacylated lipoproteins, lipoteichoic acids, and beta-glucans. TLR2 also heterodimerizes with TLR6 to bind diacylated lipopeptides. In addition, TLR2 can bind to various endogenous DAMPs, such as HSPs, HMGB1, uric acid, fibronectin, and other extracellular matrix proteins. It is also believed that TLR1 and TLR6 can heterodimerize with TLR10; however, TLR agonists recognized by this dimer have yet to be identified. TLR3 recognizes viral dsRNA as well as synthetic analogs of dsRNA, such as the ligand Poly I:C. TLR4 binds LPS in complex with the lipid A-binding proteins CD14 and myeloid differentiation protein 2, MD2, and recognizes various DAMPs. Endogenous TLR4 ligands that have been described include beta-defensin 2, fibronectin extra domain A EDA, HMGB1, Snapin, and tenascin C. TLR5 recognizes bacterial flagellin, TLR7 and TLR8 bind viral ssRNA, while TLR9 interacts with unmethylated CpG DNA from bacteria and some viruses. Based on sequence homology of the highly conserved TIR domain, an additional TLR was recently identified in mice. TLR10 is a surface receptor whose natural ligand is unknown. TLR11, TLR12, and TLR13 exist in mice but not in humans. TLR11 shows binding to Toxoplasma gondii inhibitory protein and uropathogenic Escherichia coli. The ligand for TLR12 has not been identified, while TLR13 is an endosomal receptor that recognizes VSV. See, e.g., Kaczanowska et al., J Leukoc Biol. 2013 Jun;93(6):847-63.

[0209] TLR signaling can act as a double-edged sword in cancer. Stimulation of TLRs in cancer cells can lead to tumor progression or suppression. For example, stimulation of TLR2, TLR4, and TLR7 / 8 was found to lead to tumor progression via production of immunosuppressive cytokines, increased cell proliferation, and resistance to apoptosis. R848 stimulation of pancreatic cancer cell lines overexpressing TLR7 / 8 led to increased cell proliferation and decreased chemosensitivity. On the other hand, stimulation of TLR2, TLR3, TLR4, TLR5, TLR7 / 8, and TLR9, often in combination with chemotherapy or immunotherapy, can lead to tumor suppression via different pathways. See, e.g., Grimmig et al., Int J Oncol. (2015) 47:857-66; Urban-Wojciuk et al., Front Immunol. 2019; 10:2388.

[0210] In some embodiments, the TLR agonist activates any of TLRs.

[0211] In some embodiments, the TLR agonist activates TLR1 or TLR2, optionally wherein the TLR agonist comprises triacylated lipoprotein, peptidoglycan, zymosan, and / or Pam3CSK4.

[0212] In some embodiments, the TLR agonist activates any of TLR2, TLR3, TLR4, TLR5, and TLR6, optionally wherein the TLR agonist comprises diacylated lipopeptide, heat shock protein, HMGB1, uric acid, fibronectin, and / or ECM protein.

[0213] In some embodiments, the TLR agonist activates TLR2, optionally wherein the TLR agonist comprises Pam3Cys, SMP-105, and / or CBLB612.

[0214] In some embodiments, the TLR agonist activates TLR3, optionally wherein the TLR agonist comprises dsRNA, PolyI:C, PolyICIC, Poly-IC12U, IPH302, ARNAX, and / or MPLA.

[0215] In some embodiments, the TLR agonist activates TLR4, optionally wherein the TLR agonist comprises LPS, lipoteichoic acid beta-defensin 2, fibronectin EDA, HMGB1, snapin, tenascin C, OK-432, AS04, and / or GLA-SE.

[0216] In some embodiments, the TLR agonist activates TLR5, optionally wherein the TLR agonist comprises flagellin, CBLB502, and / or M-VM3.

[0217] In some embodiments, the TLR agonist activates TLR6.

[0218] In some embodiments, the TLR agonist activates TLR7 or TLR8, optionally wherein the TLR agonist comprises ssRNA, CpG-A, poly G10, and / or poly G3.

[0219] In some embodiments, the TLR agonist activates TLR7, optionally wherein the TLR agonist comprises bistriazolyl and / or R848.

[0220] In some embodiments, the TLR agonist activates TLR8, optionally wherein the TLR agonist comprises VTX1463 and / or R848.

[0221] In some embodiments, the TLR agonist activates TLR9, optionally wherein the TLR agonist comprises unmethylated CpG DNA, CpG (e.g., CpG-7909, KSK-CpG, CpG-1826), MGN1703, dsSLIM, IMO2055, SD101, and / or ODN M362.

[0222] In some embodiments, the TLR agonist activates TLR10, optionally wherein the TLR agonist comprises Pam3CSK4.

[0223] In some embodiments, the TLR agonist activates TLR11, optionally wherein the TLR agonist comprises Toxoplasma gondii inhibin.

[0224] In some embodiments, the TLR agonist activates TLR12.

[0225] In some embodiments, the TLR agonist activates TLR13, optionally wherein the TLR agonist comprises VSV.

[0226] In some embodiments, the TLR agonist activates TLR on macrophages.

[0227] In some embodiments, the TLR agonist activates TLR1, TLR2, TLR3, TLR4, TLR7, TLR8, and / or TLR9.

[0228] In some embodiments, the TLR comprises TLR1, TLR4, and / or TLR9. In some embodiments, the TLR comprises TLR9.

[0229] In some embodiments, the TLR comprises TLR2, TLR4, TLR7, and / or TLR8.

[0230] In some embodiments, the TLR agonist comprises CpG. In some embodiments, the TLR agonist comprises poly I:C. In some embodiments, the TLR agonist comprises CpG and / or poly I:C. In some embodiments, the TLR agonist comprises CpG, poly I:C, and / or R848. In some embodiments, the TLR agonist comprises CpG, poly I:C, and R848, e.g., at a ratio of 1 : 1 : 1.

[0231] In some embodiments, the methods described herein further comprise assessing whether the individual has an ongoing infection. In some embodiments, a reduced amount of the TLR agonist is administered when the individual has an ongoing infection. In some embodiments, the TLR agonist can be avoided when the individual has an ongoing infection.

[0232] Radiation therapy

[0233] In some embodiments, the pro-inflammatory agent comprises or is radiation therapy. Radiation activates an interconnected network of cytokines, adhesion molecules, ROS / RNS, and DAMPs, resulting in a self-amplifying cascade that produces a pro-inflammatory, pro-oxidative tumor microenvironment that ultimately leads to tumor cell death. See, e.g., McKelvey et al., Mamm Genome. 2018; 29(11):843-865.

[0234] In some embodiments, the radiation therapy comprises irradiation at a site of a cancer to be treated.

[0235] In some embodiments, the radiation therapy comprises irradiation at a site different from a site of a cancer to be treated.

[0236] In some embodiments, the radiation therapy is intraoperative radiation therapy (“IORT”). In particular embodiments, the radiation is localized to a tumor site. The patient can receive intraoperative radiation before or after the tumor is resected. The tumor site can comprise different types of cells, including cancerous cells and benign cells. In certain embodiments, the radiation therapy is stereotactic body radiation therapy (“SBRT”) or stereotactic radiosurgery (“SRS”).

[0237] In some embodiments, the radiation is ionizing radiation, such as particle beam radiation. The particle beam radiation can be selected from any of an electron, a proton, a neutron, a heavy ion such as a carbon ion, or a pion. The ionizing radiation can be selected from x-rays, UV light, gamma rays, or microwaves. In some embodiments, the radiation therapy can comprise subjecting the patient to one or more types of radiation therapy.

[0238] In some embodiments, the tumor cells are sensitized to radiation using a radiosensitizer. The use of such drugs, known as radiosensitizers, provides a way to increase the radiosensitivity of tumors to radiotherapy, thereby avoiding the need to increase the radiation dose to levels that are harmful to surrounding organs and tissues. See, e.g., US9656098B2.

[0239] In some embodiments, the dose of radiation therapy is non-ablative, insufficient to eliminate the tumor (kill all tumor cells). In some embodiments, the radiation therapy is selected from the group consisting of external beam radiation therapy, internal beam therapy (brachytherapy), intraoperative radiation therapy (IORT), systemic radiation therapy, radioimmunotherapy, and administration of a radiosensitizer and a radioprotector.

[0240] In some embodiments, the radiation therapy is external beam radiation therapy, optionally including three-dimensional conformal radiation therapy (3D-RT), intensity modulated radiation therapy (IMRT), photon beam therapy, image-guided radiation therapy (IGRT), and stereotactic radiation therapy (SRT).

[0241] In some embodiments, the radiation therapy includes administration of a radiopharmaceutical. Radiopharmaceuticals can be delivered via any vehicle such as a cell, protein, or small molecule complex. In some embodiments, the radiopharmaceutical is administered to the tumor tissue. See, e.g., Sgouros et al., Radiopharmaceutical therapy in cancer: clinical advances and challenges. Nat Rev Drug Discov 19, 589-608 (2020).

[0242] In some embodiments, the radiation therapy is brachytherapy, optionally including interstitial brachytherapy, intracavitary brachytherapy, intraluminal brachytherapy, and intravenously administered radiolabeled molecules.

[0243] STING activators

[0244] In some embodiments, the pro-inflammatory agent includes or is a STING activator.

[0245] The IFN gene stimulator (STING, also known as TMEM173, MITA, MPYS, or ERIS) is a pattern recognition receptor (PRR) that recognizes cytosolic DNA in the form of cyclic dinucleotides (CDNs), such as the bacterial product cyclic guanosine monophosphate-adenosine monophosphate (3'3' cGAMP). In addition to bacterial components, other forms of DNA from viruses or host cells that find their way into the cytosol are recognized by the enzyme c-GMP-AMP (cGAMP) synthase (cGAS). Upon cytosolic DNA binding, cGAS converts ATP and GTP into the animal-specific CDN 2'3'-cGAMP for STING recognition and activation. STING is a transmembrane protein that exists as a dimer anchored within the endoplasmic reticulum membrane and forms a V-shaped pocket that binds cytosolic CDNs. Ligand binding causes a dramatic conformational change in the C-terminal domain of STING, which mediates its transport to the Golgi compartment. In the Golgi, STING recruits TANK-binding kinase 1 (TBK1) that promotes IRF3 phosphorylation, nuclear translocation, and strong induction of type I IFN (e.g., IFN-b) transcription. STING also triggers a potent proinflammatory cytokine response [e.g., tumor necrosis factor (TNF)] through activation of nuclear factor-kappa B (NF-kB), and this part of the pathway can be mediated independently of TBK1 via the closely related homolog IKK. See, e.g., Peng et al., Front Immunol. 2022 Feb 25; 13: 794776; Amougezar et al., Cancers (Basel). 2021 May 30; 13(11): 2695.

[0246] In some embodiments, the STING activator is a cyclic guanosine monophosphate-adenosine monophosphate (cGAMP, e.g., 3'3' cGAMP, e.g., 2'3' cGAMP).

[0247] In some embodiments, the STING activator is a bacterial vector (e.g., SYNB1891, STACT-TREX-1).

[0248] In some embodiments, the STING activator is a CDN compound (e.g., ADU-S100, BI-STING, BMS-986301, GSK532, JNJ-4412, MK-1454, SB11285, 3'3'-cyclic AIMP).

[0249] In some embodiments, the STING activator is a non-CDN small molecule (e.g., ALG- 031048, E7755, JNJ-‘6196, MK-2118, MSA-1, MSA-2, SNX281, SR-717, TAK676, TTI-10001).

[0250] In some embodiments, the STING activator is a nanovaccine (e.g., PC7A NP, cCAMP-NP, ONM-500).

[0251] In some embodiments, the STING activator is an antibody-drug conjugate (e.g., XMT-2056, CRD-5500).

[0252] Other exemplary STING activators can be found in Amougezar et al., Cancers (Basel). 2021 May 30; 13(11): 2695, which is incorporated by reference herein in its entirety.

[0253] PAMP / DAMP activators

[0254] In some embodiments, the pro-inflammatory agent comprises or is a PAMP / DAMP activator.

[0255] Organisms perceive microbial infection through innate receptors encoded in the genome, which are known as pattern recognition receptors, including Toll-like receptors (TLRs), nucleotide-binding and oligomerization domain (NOD)-like receptors, and retinoic acid-inducible gene I (RIG-I)-like receptors. These receptors recognize pathogen-associated molecular patterns (PAMPs) expressed by bacteria, fungi, and viruses, but also bind damage-associated molecular patterns (DAMPs), which are molecules released by sterile injury. Thus, PAMPs and DAMPs that bind to the same type of receptor initiate the same intracellular pathways, culminating in the same effector functions. See, e.g., Alisi et al., Hepatology. 2011 Nov; 54(5): 1500-2.

[0256] In some embodiments, the pro-inflammatory agent is a PAMP activator. Exemplary PAMP activators include triacyl lipopeptides, LPS, lipoproteins, peptidoglycans, zymosan, lipoteichoic acid, trypanosome phospholipids, Pam3Cys porin, lipoarabinomannan, double-stranded RNA, poly(I:C), trypanosome lipids, paclitaxel, Pseudomonas exoenzyme S, RSV F protein, MMTV envelope protein, flagellin, diacyl lipopeptides, single-stranded RNA, imiquimod, single-stranded RNA, resiquimod, bacterial / viral DNA, CpG DNA, urea bacterial, and Toxoplasma LPS.

[0257] In some embodiments, the pro-inflammatory agent is a DAMP activator. Exemplary DAMP activators include defensins, HSP60, HSP70, messenger RNA, low molecular weight hyaluronan, fibrinogen, fibronectin, fx1-defensin, heparan sulfate, HSP60, HSP70, HSP90, HMGB1, and unmethylated CpG DNA.

[0258] chemotherapeutic agent

[0259] In some embodiments, the pro-inflammatory agent comprises or is a chemotherapeutic agent.

[0260] In some embodiments, the chemotherapeutic agent is an alkylating agent. Exemplary alkylating agents include nitrogen mustards (e.g., endoxan, cyclophosphamide, ifosfamide), nitrosoureas (e.g., carmustine, lomustine), platinum analogs (e.g., carboplatin, cisplatin, oxaliplatin), triazenes (e.g., dacarbazine, procarbazine, temozolomide), alkyl sulfonates (e.g., busulfan), and ethylenimines (e.g., thiotepa).

[0261] In some embodiments, the chemotherapeutic agent is an antimetabolite. Exemplary antimetabolites include cytidine analogs (e.g., azacitidine, decitabine, cytarabine, gemcitabine), folate antagonists (e.g., methotrexate, pemetrexed), purine analogs (e.g., cladribine, clofarabine, nelarabine), pyrimidine analogs (e.g., fluorouracil (5-FU), capecitabine (a prodrug of 5-FU)).

[0262] In some embodiments, the chemotherapeutic agent is an anti-microtubule agent. Exemplary anti-microtubule agents include topoisomerase II inhibitors (e.g., anthracyclines, doxorubicin, daunorubicin, idarubicin, mitoxantrone), topoisomerase I inhibitors (e.g., irinotecan, topotecan), taxanes (e.g., paclitaxel, docetaxel, cabazitaxel), vinca alkaloids (e.g., vinblastine, vincristine, vinorelbine), antibiotics (e.g., actinomycin D, bleomycin, daunorubicin).

[0263] Other exemplary chemotherapeutic agents include hydroxyurea, retinoids, arsenic trioxide, and proteasome inhibitors (e.g., bortezomib).

[0264] pro-inflammatory cytokine

[0265] In some embodiments, the pro-inflammatory agent is a pro-inflammatory cytokine.

[0266] In some embodiments, the pro-inflammatory cytokine promotes M1 macrophages. See, e.g., Duque et al., Front Immunol. 2014; 5: 491. In some embodiments, the pro-inflammatory cytokine comprises or is TNF, IFNy, and / or GM-CSF.

[0267] In some embodiments, the pro-inflammatory cytokine comprises IL-6, TNFa, a cytokine from the IL-1 family (e.g., IL-1a, IL-1b, IL-18, IL-33, and IL-36), and / or IFNy.

[0268] In some embodiments, the pro-inflammatory cytokine comprises a cytokine from the IL-1 family. In some embodiments, the pro-inflammatory cytokine comprises any one or more of IL-1a, IL-1b, IL-18, IL-33, and IL-36. See, e.g., Sims, J., Smith, D. The IL-1 family: regulators of immunity. Nat Rev Immunol 10, 89-102 (2010).

[0269] Checkpoint inhibitors

[0270] In some embodiments, the pro-inflammatory agent is a checkpoint inhibitor. Immune checkpoints are pathways with inhibitory or stimulatory characteristics that maintain self-tolerance and assist in immune responses. The most well-described checkpoints are inhibitory in nature and include cytotoxic T-lymphocyte-associated molecule-4 (CTLA-4), programmed cell death receptor-1 (PD-1), and programmed cell death ligand-1 (PD-L1). See, e.g., Marin-Acevedo et al., J Hematol Oncol 14, 45 (2021).

[0271] In some embodiments, the checkpoint inhibitor targets CLTA-4, PD-1, or PD-L1 (e.g., an antibody that targets CTLA-4, PD-1, or PD-L1).

[0272] In some embodiments, the checkpoint inhibitor targets LAG-3, TIM-3, B7-H3, B7-H4, A2aR, CD73, NKG2A, PVRIG / PVRL2, CEACAM1, CEACAM 5 / 6, FAK, CCL2 / CCR2, LIF, CD47 / SIRPa, CSF-1 (M-CSF) / CSF-1R, IL-1 / IL-1R3 (IL-1RAP), IL-8, SEMA4D, Ang-2, CLEVER-1, Axl, or phosphatidylserine.

[0273] In some embodiments, the checkpoint inhibitor comprises or is Libivimab, Cemiplimab, Nivolumab, Pembrolizumab, Atezolizumab, Avelumab, Durvalumab, LAG525 (IMP701), REGN3767, BI 754,091, Toripalimab (MGD013), Ipilimumab alpha (IMP321), FS118, MBG453, Sym023, TSR-022, MGC018, FPA150, EOS100850, AB928, CPI-006, Monalizumab, COM701, CM24, NEO-201, Defactinib, PF-04136309, MSC-1, Hu5F9-G4 (5F9), ALX148, TTI-662, RRx-001, Lannotamab (MCS110), LY3022855, SNDX-6352, Emibetuzumab (RG 7155), Plitharam (PLX3397), CAN04, Canakinumab (ACZ885), BMS-986253, Pembinanib (VX15 / 2503), Tremikimab, FP-1305, Vynoba (EnaV), or Batiximab.

[0274] Cancer vaccine

[0275] In some embodiments, the pro-inflammatory agent comprises or is a cancer vaccine. Cancer vaccines stimulate anti-tumor immunity with tumor antigens, which can be delivered in the form of whole cells, peptides, nucleic acids, etc. An ideal cancer vaccine can overcome the immunosuppression of tumors and induce both humoral and cellular immunity.

[0276] In some embodiments, the cancer vaccine comprises a cell-based vaccine, a peptide-based vaccine, a virus-based vaccine, and / or a nucleic acid-based vaccine. See, e.g., Liu et al., JHematol Oncol 15, 28 (2022).

[0277] Cell-based vaccines are initially a form of cancer vaccines. Cell-based cancer vaccines are usually prepared from whole cells or cell fragments, which contain almost tumor antigens, inducing a more extensive antigen immune response. DC vaccines are an important branch of cell-based vaccines. DC-based personalized neoantigen cancer vaccines have shown good anti-tumor effects in clinics. Viruses are naturally immunogenic, and their genetic material can be engineered to contain sequences encoding tumor antigens. Several recombinant viruses such as adenovirus can infect immune cells as carriers. Engineered viral vaccines can present a large amount of tumor antigens in the immune system and produce anti-tumor immunity. In addition, oncolytic viruses can also be used as carriers. In addition to providing tumor antigens, the virus itself can also lyse the tumor, release tumor antigens, further improve the effectiveness of the vaccine, and produce long-term immune memory.

[0278] Peptide-based subunit vaccines, including chemically and biologically synthesized preparations of predicted or known specific tumor antigens, induce strong immune responses against specific tumor antigen sites. Peptide-based subunit vaccines in combination with adjuvants can effectively elicit humoral immune responses and are suitable for the prevention and treatment of viral infectious diseases.

[0279] HBV and HPV vaccines for liver cancer and cervical cancer are mainly peptide-based subunit vaccines. In particular, in recent years, virus-like particle (VLP)-based subunit vaccines that can activate cellular immune responses have shown good anti-tumor activity.

[0280] Nucleic acid vaccines induce strong MHC I-mediated CD8+ T cell responses; therefore, it is an ideal cancer vaccine platform. Nucleic acid vaccines can deliver multiple antigens simultaneously to trigger both humoral and cellular immunity. In addition, nucleic acid vaccines can encode full-length tumor antigens, allowing APCs to cross-present various epitopes or present several antigens simultaneously. Finally, nucleic acid vaccines are simple and fast to prepare, which are suitable for the development of personalized neoantigen cancer vaccines.

[0281] Oncolytic viruses

[0282] In some embodiments, the proinflammatory agent is an oncolytic virus (OV). Oncolytic viruses (OVs) are organisms that are able to recognize, infect, and lyse different cells in the tumor environment with the aim of stabilizing and slowing down tumor progression. They can present a natural tropism for cancer cells or be genetically directed to recognize specific targets. See, e.g., Apolonio et al., World J Virol. 2021 Sep 25; 10(5): 229-255.

[0283] Oncolytic viruses represent an exciting new approach to cancer treatment. Such viruses have the extraordinary ability to capture and terminate cancer cells without harming healthy cells, as well as the ability to enhance the immune system’s ability to recognize and terminate cancer cells. See, e.g., Cancer Cell. 2022 Aug 15; S1535-6108(22)00357-9.

[0284] In some embodiments, the oncolytic virus comprises or is an adenovirus (e.g., ONYX-15, LOAd703 virus), a parvovirus, a parvovirus (e.g., H-1PV), a vaccinia virus (VACV), a reovirus (e.g., Reolysin), or a herpes simplex virus (HSV, e.g., HSV-1, HSV-2, G207, L1BR1, HF10, T-VEC, Orien X010).

[0285] Other exemplary oncolytic viruses include JX-593, Coxsackievirus A21 (CVA21), Maraba virus or its MG1 variant, DNX2440 adenovirus, fowlpox virus, and Sendai virus.

[0286] Cells

[0287] In some embodiments, the pro-inflammatory agent comprises a cell that triggers an inflammatory factor. In some embodiments, the cell is a tumor infiltrating lymphocyte. In some embodiments, the cell specifically recognizes a tumor antigen (e.g., is engineered to express a CAR that recognizes a tumor antigen). In some embodiments, the cell is a T cell. In some embodiments, the cell is a CAR-T cell. In some embodiments, the cell is an NK cell (e.g., a CAR-NK cell). In some embodiments, the cell is a neutrophil (e.g., a CAR-expressing neutrophil). In some embodiments, the cell is a TCR-T cell. In some embodiments, the cell is an APC (e.g., a macrophage or a dendritic cell). In some embodiments, the cell is a CAR-macrophage or a CAR-monocyte. In some embodiments, the cell is a SIRPant-macrophage. In some embodiments, the cell is a stem cell. In some embodiments, the cell is allogeneic. In some embodiments, the cell is autologous.

[0288] Immune cells, monocytes, or macrophages

[0289] The immune cells described herein encompass various types of immune cells.

[0290] In some embodiments, the immune cells comprise monocytes or macrophages described herein. In some embodiments, the macrophages are identified by F4 / 80 expression. In some embodiments, the macrophages have an Ml phenotype. In some embodiments, at least 50%, 60%, 70%, 80%, 90%, 95%, 98%, or 99% of the macrophages in the immune cells have an Ml phenotype.

[0291] In some embodiments, the macrophages are engineered to be tyrosine kinase inhibitor expressing and / or activation deficient. In some embodiments, the monocytes or macrophages express a reduced level of tyrosine kinase for at least one period of time (e.g., at least 1 day, 2 days, 3 days, 4 days, or 5 days), or are resistant to activation for at least one period of time (e.g., at least 1 day, 2 days, 3 days, 4 days, or 5 days). In some embodiments, the period of time is no more than about 10 days, 9 days, 8 days, 7 days, 6 days, 5 days, 4 days, or 3 days.

[0292] In some embodiments, the monocyte or macrophage has reduced tyrosine kinase activity for no more than about 5 consecutive days (e.g., no more than 5 days, 4 days, or 3 days) before the level of tyrosine kinase activity returns to normal.

[0293] Methods of engineering monocytes or macrophages to transiently express reduced levels of tyrosine kinases are well known in the art. Exemplary methods include contacting a monocyte or macrophage, in vivo or in vitro, with a tyrosine kinase inhibitor described herein, such as a small molecule, a nucleic acid (e.g., siRNA, shRNA, antisense RNA, microRNA), a nucleic acid editing system (e.g., a CRISPR system), and a protein agent (e.g., an antibody agent that targets a tyrosine kinase or an activated tyrosine kinase).

[0294] In some embodiments, the immune cell comprises a T cell (e.g., a CAR-T cell).

[0295] In some embodiments, the immune cell comprises an NK cell (e.g., a CAR-NK cell).

[0296] In some embodiments, the immune cell comprises a neutrophil (e.g., a CAR-expressing neutrophil).

[0297] In some embodiments, the immune cell comprises an antigen presenting cell (APC, e.g., a dendritic cell).

[0298] In some embodiments, the immune cell is derived from the same individual (i.e., autologous). In some embodiments, the immune cell is allogeneic.

[0299] In some embodiments, the immune cell is engineered to express a chimeric antigen receptor, optionally wherein the chimeric antigen receptor specifically binds to a tumor antigen.

[0300] In some embodiments, the immune cell expresses high levels of MHC-I, MHC-II, CD80, and / or CD86. In some embodiments, the immune cell expresses high levels of MHC-I, MHC-II, CD80, and / or CD86 when the expression level of MHC-I, MHC-II, CD80, and / or CD86 on the immune cell is comparable (e.g., at least more than 50%) to the expression level on an activated antigen presenting cell (APC).

[0301] In some embodiments, the immune cell expresses one or more proinflammatory cytokines, optionally wherein the one or more proinflammatory cytokines comprises TNFa and / or IL-12.

[0302] In some embodiments, the immune cell does not express a significant level of TGF and / or IL-10.

[0303] In some embodiments, the tyrosine kinase inhibitor and the immune cell are administered within 24 hours (e.g., 12 hours, 8 hours, 4 hours, 2 hours, 1 hour, or 0.5 hours) of each other, optionally wherein the tyrosine kinase inhibitor and the immune cell are administered within 4 hours of each other.

[0304] In some embodiments, the above-described tyrosine kinase inhibitor, immune cell, and proinflammatory agent are administered within 24 hours (e.g., 12 hours, 8 hours, 4 hours, 2 hours, 1 hour, or 0.5 hours) of each other. In some embodiments, the immune cell is administered simultaneously or concurrently with the tyrosine kinase inhibitor and / or the proinflammatory agent.

[0305] Inflammatory response or ongoing infection

[0306] There is substantial evidence that both acute and chronic inflammation are associated with the development and progression of cancer. Advances in inflammation research have revealed links between inflammatory processes and tumor transformation, tumor progression, and the development of metastasis and recurrence. Moreover, tumor invasive procedures (surgery and biopsies) affect the remaining tumor cells by increasing their survival, proliferation, and migration. One of the concepts that explains this phenomenon is the induction of a wound healing response. While in normal tissues tissue repair is necessary, in tumor tissues the induction of adaptive and innate immune responses associated with wound healing stimulates tumor cell survival, angiogenesis, and extravasation of circulating tumor cells. See, e.g., Singh et al., Ann Afr Med. 2019 Jul-Sep; 18(3): 121-126; Piotrowski et al., Rep Pract Oncol Radiother. 2020 May-Jun; 25(3):422-427.

[0307] However, as demonstrated in this application, the combined use of a tyrosine kinase inhibitor and a proinflammatory agent releases a proinflammatory response and transforms the immunosuppressive tumor environment into a site with inflammatory markers. See, e.g., FIG. 7F. A significant anti-tumor effect is achieved. These results provide support for the use of the methods described herein to treat individuals in an inflammatory response.

[0308] In some embodiments, the individual is in an inflammatory response or has an ongoing infection when treated with the methods described herein. The inflammatory response described herein can be reflected in, for example, a) an increase in one or more (e.g., at least one, two, three, four, five) inflammatory cytokines (such as IFNy, IL-12b, TNFa, IL-6, IL- lb, IFN-al, IFN-a2, IFN-bl), b) a decrease in one or more (e.g., at least one, two, or three) anti-inflammatory cytokines (such as TGFbl, TGFb2, TGFb3), c) an increase in infiltrating immune cells (such as T cells, NK cells, macrophages, neutrophils), d) a decrease in suppressive immune cells (such as MDSCs), and / or e) an increase in one or more (e.g., at least one, two, three, four, or five) immunogenic costimulatory molecules (such as CD80, CD86, OX40L, CD40, ICOS-L, PD-L1, GITRL) in a tissue (e.g., tumor tissue) or immune cell (such as a macrophage).

[0309] In some embodiments, the inflammatory response is an acute inflammatory response.

[0310] In some embodiments, the inflammatory response is in a tumor. In some embodiments, the inflammatory response is at a site different from a tumor.

[0311] In some embodiments, there is an inflammatory response, wherein there is at least two (e.g., two, three, four, or five events selected from the group consisting of) a) an increase in one or more (e.g., at least one, two, three, four, five) inflammatory cytokines (such as IFNy, IL-12b, TNFa, IL-6, IL- lb, IFN-al, IFN-a2, IFN-bl), b) a decrease in one or more (e.g., at least one, two, or three) anti-inflammatory cytokines (such as TGFbl, TGFb2, TGFb3), c) an increase in infiltrating immune cells (such as T cells, NK cells, macrophages, neutrophils), d) a decrease in suppressive immune cells (such as MDSCs), and / or e) an increase in one or more (e.g., at least one, two, three, four, or five) immunogenic costimulatory molecules (such as CD80, CD86, OX40L, CD40, ICOS-L, PD-L1, GITRL) in a tissue (e.g., tumor tissue) or immune cell (such as a macrophage).

[0312] In some embodiments, an increase described herein refers to an amount of a level that is increased by at least about 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, or 200% compared to a reference state, optionally wherein the reference state is a state when the individual is neither treated with a method described herein nor infected with a pathogen. In some embodiments, an increase described herein refers to an amount of a level that is increased by at least about 5-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, 100-fold, 150-fold, 200-fold, 250-fold, 500-fold, or 1000-fold compared to a reference state, optionally wherein the reference state is a state when the individual is neither treated with a method described herein nor infected with a pathogen. In some embodiments, the reference state is a state when a healthy individual is not infected with a pathogen.

[0313] In some embodiments, a decrease described herein refers to an amount of a level that is decreased by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 99.9% compared to a reference state, optionally wherein the reference state is a state when the individual is neither treated with a method described herein nor infected with a pathogen. In some embodiments, the reference state is a state when a healthy individual is not infected with a pathogen.

[0314] In some embodiments, the individual has an inflammatory response (e.g., in a tumor, e.g., at a site other than a tumor) within about one week, 6 days, 5 days, 4 days, 3 days, 2 days, or one day before and / or after administration of the tyrosine kinase inhibitor.

[0315] In some embodiments, the individual has an ongoing inflammatory response (e.g., in a tumor, e.g., at a site other than a tumor) when the tyrosine kinase inhibitor is administered.

[0316] In some embodiments, the individual has an ongoing infection when the tyrosine kinase inhibitor is administered. In some embodiments, the method further comprises assessing the individual for the presence of an infection (e.g., an infection associated with a virus, fungus, and / or bacteria).

[0317] Immunogenic cell death

[0318] In some embodiments, the individual has immunogenic cell death when treated with a method described herein.

[0319] Immuno genic cell death (ICD) is a type of cancer cell death that can be induced by different stressors, including but not limited to (1) intracellular pathogens; (2) conventional chemotherapeutics such as anthracyclines, DNA-damaging agents, and proteasome inhibitors; (3) targeted anticancer agents such as the tyrosine kinase inhibitor crizotinib, the epidermal growth factor receptor-specific monoclonal antibody cetuximab, and the poly-ADP ribose polymerase (PARP) inhibitors; and (4) many physical therapies, including hypericin- and radoperoxyl-based photodynamic therapy, extracorporeal photochemotherapy, various forms of ionizing radiation, high hydrostatic pressure, and severe heat shock. It involves activation of the immune system against cancer in an immunocompetent host. ICD includes release of damage-associated molecular patterns (DAMPs) from dying tumor cells, which leads to activation of tumor-specific immune responses, thus potentiating long-term efficacy of anticancer drugs by combining direct cancer cell killing with antitumor immunity. DAMPs include cell surface exposure of calreticulin (CRT) and heat shock proteins (HSP70 and HSP90), extracellular release of adenosine triphosphate (ATP), high mobility group protein B1 (HMGB1), type I IFN, and IL-1 cytokine family members. See, e.g., Ahmed et al., Mol Oncol. 2020 Dec;14(12):2994-3006 and Fucikova et al., Cell Death Dis. 2020 Nov 26;11(11):1013.

[0320] Key DAMPs for which cell death is considered immunogenic include calreticulin, high mobility group protein B1 (HMGB1), ATP, annexin A1 (ANXA1), and type I IFN. The main hallmarks of immunogenic cell death (ICD) can be assessed by flow cytometry, (immuno)fluorescence microscopy, immunoblotting, or luminescence analysis based on various different approaches. See, e.g., Cell Death Dis. 2020 Nov 26;11(11):1013.

[0321] In some embodiments, the individual has ICD (e.g., in the tumor, e.g., at a site other than the tumor) within about one week, 6 days, 5 days, 4 days, 3 days, 2 days, or one day before and / or after administration of the tyrosine kinase inhibitor.

[0322] In some embodiments, the individual has ongoing ICD (e.g., in the tumor, e.g., at a site other than the tumor) when the tyrosine kinase inhibitor is administered.

[0323] In some embodiments, the individual has ICD when a sample from the cancer has a higher level of one or more (e.g., at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% higher) DAMP than a reference sample (e.g., a corresponding sample in a healthy control, e.g., a sample from the cancer prior to administration of an ICD-inducing treatment). In some embodiments, the DAMP is selected from the group consisting of endoplasmic reticulum (ER) chaperone molecules (e.g., calreticulin (CALR), e.g., heat shock proteins (HSPs)), non-histone chromatin-binding protein high-mobility group box 1 (HMGB1), cytoplasmic protein annexin A1 (ANXA1), and small metabolites ATP and type I interferon (IFN).

[0324] The individual

[0325] In some embodiments, the individual has a solid tumor. In some embodiments, the individual has a hematological cancer.

[0326] In some embodiments, the individual has an advanced cancer. In some embodiments, the individual has an end-stage cancer. In some embodiments, the individual has a malignant cancer. In some embodiments, the individual has a cancer that is at stage II, III, or IV. In some embodiments, the individual has a non-surgical tumor and / or metastasis. In some embodiments, the individual is an end-stage individual.

[0327] In some embodiments, the individual has received a treatment that induces an inflammatory response or immunogenic cell death (e.g., radiation therapy) (e.g., within 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, 16 hours, 20 hours, or 24 hours prior to administration of the tyrosine kinase inhibitor, e.g., within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 7 days prior to administration of the tyrosine kinase inhibitor). In some embodiments, the individual will receive a treatment that induces an inflammatory response or immunogenic cell death (e.g., radiation therapy) (e.g., within 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, 16 hours, 20 hours, or 24 hours after administration of the tyrosine kinase inhibitor, e.g., within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 7 days after administration of the tyrosine kinase inhibitor).

[0328] In some embodiments, the individual has received a proinflammatory agent (such as any of the proinflammatory agents described herein) (e.g., within 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, 16 hours, 20 hours, or 24 hours prior to administration of the tyrosine kinase inhibitor, e.g., within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 7 days prior to administration of the tyrosine kinase inhibitor). In some embodiments, the individual will receive a proinflammatory agent (such as any of the proinflammatory agents described herein) (e.g., within 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, 16 hours, 20 hours, or 24 hours after administration of the tyrosine kinase inhibitor, e.g., within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 7 days after administration of the tyrosine kinase inhibitor).

[0329] In some embodiments, the individual does not have an autoimmune disease.

[0330] In some embodiments, the individual is a female. In some embodiments, the individual is a male.

[0331] In some embodiments, the individual is a human. In some embodiments, the individual is at least about 50 years old, 55 years old, 60 years old, 65 years old, 70 years old, or 75 years old.

[0332] In some embodiments, the individual is selected for treatment based on a high expression level and / or a high activation level of the tyrosine kinase in tumor tissue. In some embodiments, the individual has a high expression level and / or a high activation level of the tyrosine kinase when the expression level and / or activation level is at least about 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, or 200% higher than a reference expression level and / or a reference activation level of the tyrosine kinase. In some embodiments, the individual has a high expression level and / or a high activation level of the tyrosine kinase when the expression level and / or activation level is at least about 5-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, 100-fold, 150-fold, 200-fold, 250-fold, 500-fold, or 1000-fold higher than a reference expression level and / or a reference activation level of the tyrosine kinase. In some embodiments, the reference expression level or reference activation level of the tyrosine kinase is the corresponding expression or activation level of the tyrosine kinase in a reference state, in which the individual is not treated with a proinflammatory agent (or any immunotherapy).

[0333] In some embodiments, the individual is at risk of developing systemic inflammation and / or CRS. In some embodiments, the individual develops systemic inflammation and / or CRS prior to administration of an agent that reduces systemic inflammation (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody). Cytokine release syndrome can impair or cause organ failure in most organ systems. For example, organs that can be impaired due to CRS can include, but are not limited to, the lungs, kidneys, liver, brain, heart, spleen, or any combination thereof, e.g., multi-organ failure.

[0334] In some embodiments, the individual is administered an agent that reduces systemic inflammation (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody). In some embodiments, administration occurs prior to the individual developing systemic inflammation. In some embodiments, the individual develops mild cytokine release syndrome. In some embodiments, the individual develops Grade 1 CRS. Mild symptoms of CRS can include fever, fatigue, headache, rash, arthralgias, and myalgias. Mild CRS can be treated by treating symptoms or by administration of anti-inflammatory drugs such as corticosteroids. Mild CRS can typically be resolved within one to two weeks and does not require hospitalization.

[0335] In some embodiments, the individual does not develop severe cytokine release syndrome. In some embodiments, the individual does not develop Grade 2 CRS. In some embodiments, the individual does not develop Grade 3 CRS. In some embodiments, the individual does not develop Grade 4 CRS. More severe cases are characterized by low blood pressure and high fever, and severe CRS can progress to an uncontrolled systemic inflammatory response with circulatory shock requiring vasopressors, vascular leakage, disseminated intravascular coagulation, and multi-organ system failure. More severe CRS cases typically require hospitalization for treatment of symptoms. Common laboratory abnormalities in patients with CRS include cytopenias, elevations in creatinine and liver enzymes, derangements in coagulation parameters, and high CRP. As shown in Table 1 below, there are currently four grading systems for cytokine release syndrome. See, e.g., Liu, D., and Zhao, J., J Hematol Oncol. 2018 Sep 24; 11(1): 121; and Shimabukuro-Vornhagen, A., et al., J Immunother Cancer. 2018 Jun 15; 6(1): 56, which are hereby incorporated by reference in their entireties.

[0336] In some embodiments, the individual has developed CRS prior to administration of the agent that reduces systemic inflammation (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody). In some embodiments, the individual has developed Grade 1 CRS. In some embodiments, the individual has developed Grade 2 CRS. In some embodiments, the individual has developed Grade 3 CRS. In some embodiments, the individual has developed Grade 4 CRS. In some embodiments, the agent that reduces systemic inflammation (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) is administered to the individual who has developed CRS. In some embodiments, the agent that reduces systemic inflammation (e.g., a TNFa inhibitor, e.g., an anti-TNFa antibody) ameliorates, eliminates, or reverses CRS, including organ damage, e.g., proinflammatory organ damage (e.g., nephritis, hepatitis, pneumonitis, myocarditis, appendicitis).

[0337] Table 1. Cytokine Release Syndrome Medical Grading System.

[0338]

[0339]

[0340]

[0341] In some embodiments, the individual does not develop a cytokine storm. In some embodiments, the individual develops a mild cytokine storm. In some embodiments, the individual does not develop a severe or life-threatening cytokine storm. Cytokine storm appears to be primarily a result of non-specific T cell activation, whereas CRS is more often a direct result of antigen-specific T cell activation. The clinical presentation of cytokine storm and CRS can be similar (Liu, D. and Zhao, J., J Hematol Oncol. 2018 Sep 24; 11(1): 121).

[0342] Cancer

[0343] The cancer described herein can be any type or kind. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a hematological cancer.

[0344] In some embodiments, the cancer is an advanced cancer. In some embodiments, the cancer is an end-stage cancer. In some embodiments, the cancer is a terminal-stage cancer. In some embodiments, the cancer is in Stage II, Stage III, or Stage IV. In some embodiments, the cancer is an inoperable tumor and / or is malignant.

[0345] In some embodiments, the length of the tumor is at least 0.2 cm, 0.4 cm, 0.6 cm, 0.8 cm, 1 cm, 2 cm, 3 cm, 4 cm, or 5 cm.

[0346] Examples of cancer described herein include, but are not limited to, adrenocortical carcinoma, agnogenic myeloid metaplasia, AIDS-related cancers (e.g., AIDS-related lymphoma), anal cancer, appendix cancer, astrocytomas (e.g., cerebellum and brain), basal cell carcinoma, biliary cancer (e.g., extrahepatic), bladder cancer, bone cancer (osteosarcoma and malignant fibrous histiocytoma), brain tumors (e.g., glioma, brain stem glioma, cerebellum or brain astrocytoma (e.g., pilocytic astrocytoma, diffuse astrocytoma, anaplastic (malignant) astrocytoma), malignant glioma, ependymal tumor, oligodendroglioma, meningioma, craniopharyngioma, hemangioblastoma, medulloblastoma, supratentorial primitive neuroectodermal tumor, visual pathway and hypothalamic glioma, and glioblastoma), breast cancer, bronchial adenomas / carcinoids, carcinoid tumors (e.g., gastrointestinal carcinoid tumors), carcinoma of unknown primary, central nervous system lymphoma, cervical cancer, colon cancer, colorectal cancer, chronic myeloproliferative disorders, endometrial cancer (e.g., uterine cancer), ependymal tumor, esophageal cancer, Ewing's family of tumors, eye cancer (e.g., intraocular melanoma and retinoblastoma), gallbladder cancer, gastric (stomach) cancer, gastrointestinal carcinoid tumors, gastrointestinal stromal tumors (GIST), germ cell tumors (e.g., extracranial, extragonadal, ovarian), gestational trophoblastic tumor, head and neck cancer, hepatocellular (liver) cancer (e.g., hepatoma and hepatocellular carcinoma), hypopharyngeal cancer, islet cell carcinoma (endocrine pancreas), laryngeal cancer, laryngeal cancer, leukemia, lip and oral cavity cancer, oral cancer, liver cancer, lung cancer (e.g., small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma, and lung squamous carcinoma), lymphatic tumors (e.g., lymphoma), medulloblastoma, melanoma, mesothelioma, metastatic squamous neck cancer, mouth cancer, multiple endocrine neoplasia syndrome, myelodysplastic syndrome, myelodysplastic / myeloproliferative diseases, nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, neuroendocrine cancer, oropharyngeal cancer, ovarian cancer (e.g., ovarian epithelial cancer, ovarian germ cell tumor, ovarian low malignant potential tumor), pancreatic cancer, parathyroid cancer, penile cancer, peritoneal cancer, pharyngeal cancer, pheochromocytoma, pineal astrocytic tumors and supratentorial primitive neuroectodermal tumors, pituitary tumor, pleuropulmonary blastoma, lymphoma, primary central nervous system lymphoma (small glial cell tumor), pulmonary lymphangioleiomyomatosis, rectal cancer, renal cancer, renal pelvis and ureter cancer (transitional cell cancer), rhabdomyosarcoma, salivary gland cancer, skin cancer (e.g., non-melanoma (e.g.,squamous cell carcinoma), melanoma, and Merkel cell carcinoma, small bowel cancer, squamous cell carcinoma, testicular cancer, throat cancer, thymoma and thymic carcinoma, thyroid cancer, tuberous sclerosis, urethral cancer, vaginal cancer, vulvar cancer, Wilms' tumor, and post-transplant lymphoproliferative disorder (PTLD), abnormal vascular proliferation associated with nevi, edema (such as that associated with brain tumors), and Meigs' syndrome.

[0347] In some embodiments, the cancer is a virus infection-associated cancer. In some embodiments, the cancer is a human papillomavirus (HPV)-associated cancer (e.g., HPV-associated cervical cancer, e.g., HPV-associated head and neck cancer, e.g., HPV-associated squamous cell carcinoma). In some embodiments, the cancer is a human herpesvirus 8 (HHV8)-associated cancer (e.g., Kaposi sarcoma). In some embodiments, the cancer is a human T-lymphotropic virus (HTLV-1)-associated cancer (e.g., adult T-cell leukemia or lymphoma). In some embodiments, the cancer is an Epstein-Barr virus (EBV)-associated cancer (e.g., Burkitt’s lymphoma, Hodgkin and non-Hodgkin lymphoma, gastric cancer). In some embodiments, the cancer is a hepatitis B virus (HBV)-associated cancer (e.g., liver cancer). In some embodiments, the cancer is a hepatitis C virus)-associated cancer (e.g., liver cancer, non-Hodgkin lymphoma).

[0348] In some embodiments, the cancer is a liver cancer, a kidney cancer, an endometrial cancer, a thymic epithelioma, a lung cancer, a spindle cell sarcoma, a chondrosarcoma, a uterine smooth muscle, a colon cancer, or a pancreatic cancer.

[0349] In some embodiments, the cancer has received and / or has not received one or more prior treatments (e.g., immune checkpoint blockade treatment (e.g., PD-1 antibody), chemotherapy, surgery, cell therapy (e.g., allogeneic NK cell infusion therapy)).

[0350] In some embodiments, the cancer is a relapsed or refractory cancer.

[0351] In some embodiments, the cancer is refractory to one or more of radiation therapy, chemotherapy, or immunotherapy (e.g., checkpoint blockade).

[0352] Dosage, methods of administration, and delivery vehicles

[0353] The tyrosine kinase inhibitors, proinflammatory agents, and immune cells (e.g., monocytes / macrophages) described herein can be administered at any desired dosage. Exemplary dosing regimens are described, e.g., in the “Tyrosine Kinase Inhibitors” section.

[0354] In some aspects, the dosage size of the pro-inflammatory agent, tyrosine kinase inhibitor, and / or immune cells (e.g., monocytes / macrophages) is determined based on one or more criteria, such as the disease burden of the subject, such as tumor burden, volume, size, or extent, range, or type of metastasis, stage, and / or likelihood or incidence of the subject developing a toxic outcome, e.g., CRS, macrophage activation syndrome, tumor lysis syndrome, neurotoxicity, and / or host immune response against the administered activated immune cells. For example, in some aspects, the number of monocytes or macrophages administered at the dosage is determined based on the tumor burden present in the subject immediately prior to beginning administration of the dosage of cells.

[0355] The pro-inflammatory agent, tyrosine kinase inhibitor, and / or immune cells (e.g., monocytes / macrophages) can be administered by any suitable means, e.g., by bolus infusion, by injection, e.g., intravenously or subcutaneously. In some embodiments, the pro-inflammatory agent, tyrosine kinase inhibitor, and / or monocytes or macrophages are administered systemically (e.g., intravenously, subcutaneously, or intraperitoneally). In some embodiments, the pro-inflammatory agent, tyrosine kinase inhibitor, and / or monocytes or macrophages are administered locally (e.g., intratumorally).

[0356] In some embodiments, the pro-inflammatory agent, tyrosine kinase inhibitor, and / or immune cells (e.g., monocytes / macrophages) are administered parenterally, intrapulmonarily, and intranasally, and, if local treatment is required, intralesionally or intratumorally. Parenteral infusions include intramuscular, intravenous, intra-arterial, intraperitoneal, or subcutaneous administration. In some embodiments, the pro-inflammatory agent and / or tyrosine kinase inhibitor are administered orally.

[0357] In some embodiments, the immune cells (e.g., monocytes / macrophages) and the pro-inflammatory agent are administered simultaneously. In some embodiments, the monocytes or macrophages and the pro-inflammatory agent are administered concurrently. In some embodiments, the immune cells (e.g., monocytes / macrophages) and the pro-inflammatory agent are administered sequentially. In some embodiments, the immune cells (e.g., monocytes / macrophages) and the pro-inflammatory agent are administered within about 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or 1 day. In some embodiments, the immune cells (e.g., monocytes / macrophages) and the pro-inflammatory agent are administered within about 24 hours, 16 hours, 12 hours, 8 hours, 4 hours, 2 hours, or 1 hour. In some embodiments, the immune cells (e.g., monocytes / macrophages) and the pro-inflammatory agent are administered within 30 minutes.

[0358] In some embodiments, the tyrosine kinase inhibitor and the proinflammatory agent are administered simultaneously. In some embodiments, the tyrosine kinase inhibitor and the proinflammatory agent are administered concurrently. In some embodiments, the tyrosine kinase inhibitor and the proinflammatory agent are administered sequentially. In some embodiments, the tyrosine kinase inhibitor and the proinflammatory agent are administered within about 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or 1 day. In some embodiments, the tyrosine kinase inhibitor and the proinflammatory agent are administered within about 24 hours, 16 hours, 12 hours, 8 hours, 4 hours, 2 hours, or 1 hour. In some embodiments, the tyrosine kinase inhibitor and the proinflammatory agent are administered within 30 minutes.

[0359] It is also contemplated that the tyrosine kinase inhibitors and / or proinflammatory agents described herein can be delivered via any suitable vehicle or method. In some embodiments, the tyrosine kinase inhibitors and / or proinflammatory agents are delivered directly into the tumor tissue. Different carrier systems can be used for this purpose. See, e.g., Manzari et al., Targeted drug delivery strategies for precision medicines. Nat Rev Mater 6, 351-370 (2021); Tewabe et al., J Multidiscip Healthc. 2021; 14: 1711-1724. In some embodiments, the tyrosine kinase inhibitors and / or proinflammatory agents are delivered via nanoparticles. In some embodiments, the tyrosine kinase inhibitors and / or proinflammatory agents are delivered via a controlled release system. In some embodiments, the tyrosine kinase inhibitors and / or proinflammatory agents are delivered via a biomaterial-implanted scaffold. In some embodiments, the tyrosine kinase inhibitors and / or proinflammatory agents are delivered via an injectable biomaterial scaffold. In some embodiments, the tyrosine kinase inhibitors and / or proinflammatory agents are delivered via a transdermal delivery system. See, e.g., Riley et al., Nat Rev Drug Discov. 2019 Mar; 18(3): 175-196.

[0360] In some embodiments, the tyrosine kinase inhibitor and / or pro-inflammatory agent is delivered by a cell. See, e.g., Millian et al., Ther Deliv. 2012 Jan;3(l):25-41. In some embodiments, the cell comprises a macrophage. See, e.g., Visser et al., Front Pharmacol. 2019 Jan 25;10:22. In some embodiments, the cell comprises a polymer-encapsulated human retinal pigment epithelial (aRPE) cell. See, e.g., Nash et al., Clin Cancer Res. 2022 Aug 22; CCR-22-1493. In some embodiments, the cell is encapsulated in a biocompatible material (e.g., the biocompatible alginate capsules discussed by Nash et al.)

[0361] In some embodiments, the tyrosine kinase inhibitor and / or pro-inflammatory agent is associated with an antibody construct. In some embodiments, the tyrosine kinase inhibitor and / or pro-inflammatory agent is linked to the antibody construct via a linker (e.g., a cleavable linker). In some embodiments, the antibody construct specifically recognizes a tumor-associated antigen. In some embodiments, the antibody construct comprises an antibody that recognizes a tumor antigen. In some embodiments, the antibody construct is an antibody drug conjugate (ADC).

[0362] In some embodiments, the tyrosine kinase inhibitor and / or pro-inflammatory agent is delivered via a method or device that facilitates delivery into a particular organ (e.g., an organ having a tumor). See, e.g., examples of such methods or devices in Alsaggar et al., J Drug Target. 2018 Jun-Jul; 26(5-6):385-397; Zhao et al., Cell. 2020 Apr 2; 181(l):151-167, which are incorporated by reference in their entireties.

[0363] In embodiments, the tyrosine kinase inhibitor is delivered via a controlled drug delivery system (e.g., a slow release system or vehicle, e.g., a sustained release system or vehicle). Examples of such systems can be found in, e.g., Adepu et al., Molecules. 2021 Oct; 26(19):5905; Oh et al., Chem. Asian J. 2022, 17, e202200333, which are incorporated by reference in their entireties.

[0364] VI. Compositions comprising a tyrosine kinase inhibitor

[0365] The present application also provides compositions (e.g., pharmaceutical compositions) comprising a tyrosine kinase inhibitor, a pro-inflammatory agent, and / or an immune cell for use in the above-described treatments.

[0366] In some embodiments, a composition (e.g., a pharmaceutical composition) comprising a tyrosine kinase inhibitor and a proinflammatory agent (such as any of the proinflammatory agents described herein) is provided. In some embodiments, the composition further comprises an immune cell (such as a monocyte or macrophage described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier. In some embodiments, the composition further comprises a SHP-1 inhibitor (such as TPI-1 or an analog or derivative thereof).

[0367] In some embodiments, a composition (e.g., a pharmaceutical composition) comprising a tyrosine kinase inhibitor and a TLR agonist (e.g., CpG, polyI:C, and / or R848) is provided. In some embodiments, the composition further comprises an immune cell (such as a monocyte or macrophage described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier. In some embodiments, the composition further comprises a SHP-1 inhibitor (such as TPI-1 or an analog or derivative thereof).

[0368] In some embodiments, a composition (e.g., a pharmaceutical composition) comprising a tyrosine kinase inhibitor and a STING activator (e.g., cGAMP, e.g., 2'3'-cGAMP, e.g., 3'3'-cGAMP) is provided. In some embodiments, the composition further comprises an immune cell (such as a monocyte or macrophage described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier. In some embodiments, the composition further comprises a SHP-1 inhibitor (such as TPI-1 or an analog or derivative thereof).

[0369] In some embodiments, a composition (e.g., a pharmaceutical composition) comprising a tyrosine kinase inhibitor and a chemotherapeutic agent (e.g., azathioprine (AZA), e.g., gemcitabine) is provided. In some embodiments, the composition further comprises an immune cell (such as a monocyte or macrophage described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier. In some embodiments, the composition further comprises a SHP-1 inhibitor (such as TPI-1 or an analog or derivative thereof).

[0370] In some embodiments, a composition (e.g., a pharmaceutical composition) comprising a tyrosine kinase inhibitor and a proinflammatory cytokine (e.g., IL-1b, IL-18, IL-6, and / or TNFa) is provided. In some embodiments, the composition further comprises an immune cell (such as a monocyte or macrophage described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier. In some embodiments, the composition further comprises a SHP-1 inhibitor (such as TPI-1 or an analog or derivative thereof).

[0371] In some embodiments, a composition (e.g., a pharmaceutical composition) comprising a tyrosine kinase inhibitor and a checkpoint inhibitor (e.g., an anti-PD-Ll antibody, an anti-PD-1 antibody, or an anti-CLTA4 antibody) is provided. In some embodiments, the composition further comprises an immune cell, such as a monocyte or macrophage described herein. In some embodiments, the composition further comprises a pharmaceutically acceptable carrier. In some embodiments, the composition further comprises a SHP-1 inhibitor, such as TPI-1 or an analog or derivative thereof.

[0372] In some embodiments, a composition (e.g., a pharmaceutical composition) comprising a tyrosine kinase inhibitor and a bacterial component (e.g., LPS) is provided. In some embodiments, the composition further comprises a pharmaceutically acceptable carrier. In some embodiments, the composition further comprises an immune cell, such as a monocyte or macrophage described herein. In some embodiments, the composition further comprises a pharmaceutically acceptable carrier. In some embodiments, the composition further comprises a SHP-1 inhibitor, such as TPI-1 or an analog or derivative thereof.

[0373] In some embodiments, a composition (e.g., a pharmaceutical composition) comprising a tyrosine kinase inhibitor and an agent that promotes immunogenic cell death (ICD) is provided. In some embodiments, the composition further comprises an immune cell, such as a monocyte or macrophage described herein. In some embodiments, the composition further comprises a pharmaceutically acceptable carrier. In some embodiments, the composition further comprises a SHP-1 inhibitor, such as TPI-1 or an analog or derivative thereof.

[0374] In some embodiments, a composition (e.g., a pharmaceutical composition) comprising a tyrosine kinase inhibitor and an agent for use in radiotherapy, such as any of the radiotherapies described herein, is provided. In some embodiments, the composition further comprises an immune cell, such as a monocyte or macrophage described herein. In some embodiments, the composition further comprises a pharmaceutically acceptable carrier. In some embodiments, the composition further comprises a SHP-1 inhibitor, such as TPI-1 or an analog or derivative thereof.

[0375] In some embodiments, a composition (e.g., a pharmaceutical composition) comprising a tyrosine kinase inhibitor and a PAMP / DAMP activator, such as any of the PAMP / DAMP activators described herein, is provided. In some embodiments, the composition further comprises an immune cell, such as a monocyte or macrophage described herein. In some embodiments, the composition further comprises a pharmaceutically acceptable carrier. In some embodiments, the composition further comprises a SHP-1 inhibitor, such as TPI-1 or an analog or derivative thereof.

[0376] In some embodiments, a composition (e.g., a pharmaceutical composition) comprising a tyrosine kinase inhibitor and a cancer vaccine (such as any of the cancer vaccines described herein) is provided. In some embodiments, the composition further comprises an immune cell (such as a monocyte or macrophage described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier. In some embodiments, the composition further comprises a SHP-1 inhibitor (such as TPI-1 or an analog or derivative thereof).

[0377] In some embodiments, a composition (e.g., a pharmaceutical composition) comprising a tyrosine kinase inhibitor and an oncolytic virus (such as any of the oncolytic viruses described herein) is provided. In some embodiments, the composition further comprises an immune cell (such as a monocyte or macrophage described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier. In some embodiments, the composition further comprises a SHP-1 inhibitor (such as TPI-1 or an analog or derivative thereof).

[0378] In some embodiments, a composition (e.g., a pharmaceutical composition) comprising a tyrosine kinase inhibitor and an agent for sound therapy (such as any of the sound therapies described herein) is provided. In some embodiments, the composition further comprises an immune cell (such as a monocyte or macrophage described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier. In some embodiments, the composition further comprises a SHP-1 inhibitor (such as TPI-1 or an analog or derivative thereof).

[0379] In some embodiments, a composition (e.g., a pharmaceutical composition) comprising a tyrosine kinase inhibitor and an agent for magnetotherapy (such as any of the magnetotherapies described herein) is provided. In some embodiments, the composition further comprises an immune cell (such as a monocyte or macrophage described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier. In some embodiments, the composition further comprises a SHP-1 inhibitor (such as TPI-1 or an analog or derivative thereof).

[0380] In some embodiments, a composition (e.g., a pharmaceutical composition) comprising a tyrosine kinase inhibitor and an agent for electrical or electrochemical therapy (such as any of the electrical or electrochemical therapies described herein) is provided. In some embodiments, the composition further comprises an immune cell (such as a monocyte or macrophage described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier. In some embodiments, the composition further comprises a SHP-1 inhibitor (such as TPI-1 or an analog or derivative thereof).

[0381] In some embodiments, a composition (e.g., a pharmaceutical composition) comprising a tyrosine kinase inhibitor and an agent for use in electrostatic therapy (such as any of the electrostatic therapies described herein) is provided. In some embodiments, the composition further comprises an immune cell (such as a monocyte or macrophage described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier. In some embodiments, the composition further comprises a SHP-1 inhibitor (such as TPI-1).

[0382] Exemplary Embodiments

[0383] 1. A method of treating cancer in an individual comprising administering to the individual a) a tyrosine kinase inhibitor, and b) a pro-inflammatory agent.

[0384] 2. The method of embodiment 1, wherein the method comprises systemically administering the tyrosine kinase inhibitor.

[0385] 3. The method of embodiment 1 or 2, wherein the pro-inflammatory agent comprises an agent selected from the group consisting of a TLR agonist, a STING activator, a PAMP / DAMP activator, a chemotherapy, a pro-inflammatory cytokine, a cancer vaccine, a bacterial component, a sound therapy, a magnetotherapy, an electric therapy, and an electrostatic therapy.

[0386] 4. A method of treating cancer in an individual comprising administering to the individual a tyrosine kinase inhibitor, wherein the individual is in an inflammatory response.

[0387] 5. The method of any one of embodiments 1 to 4, wherein the method further comprises intermittently administering the tyrosine kinase inhibitor to the individual.

[0388] 6. The method of embodiment 5, wherein the method comprises administering the tyrosine kinase inhibitor is administered at least three times.

[0389] 7. The method of embodiment 5 or embodiment 6, wherein the method comprises administering the tyrosine kinase inhibitor at least two times at intervals of no more than once every three days.

[0390] 8. The method of any one of embodiments 5 to 7, wherein the method comprises administering the tyrosine kinase inhibitor to the individual for at least two cycles, wherein each cycle has from about three days to about twenty days.

[0391] 9. The method of any one of embodiments 1 to 8, wherein the tyrosine kinase inhibitor inhibits SHP-1 signaling.

[0392] 10. The method of any one of embodiments 1 to 9, wherein the tyrosine kinase inhibitor has a half-life of no more than about 5 days, optionally the tyrosine kinase inhibitor has a half-life of no more than about 3 days.

[0393] 11. The method of any one of embodiments 1 to 10, wherein the tyrosine kinase inhibitor is effective to inhibit more than 50% of tyrosine kinase activity for no more than about 5 days, optionally wherein the tyrosine kinase inhibitor is effective to inhibit more than 50% of the tyrosine kinase activity for no more than about 3 days.

[0394] 12. The method of any one of embodiments 1 to 11, wherein the tyrosine kinase inhibitor is selected from the group consisting of small molecules, nucleic acids (e.g., siRNA, shRNA, antisense RNA, microRNA), nucleic acid editing systems (e.g., CRISPR system), and protein agents (e.g., antibody agents targeting tyrosine kinases or activated tyrosine kinases).

[0395] 13. The method of embodiment 12, wherein the tyrosine kinase inhibitor inhibits any one or more of: Src, Syk, Hck, Lck, Lyn, and Yes.

[0396] 14. The method of embodiment 13, wherein the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, and R406.

[0397] 15. The method of any one of embodiments 1 to 14, wherein the tyrosine kinase inhibitor is an inhibitor of a Src family tyrosine kinase.

[0398] 16. The method of any one of embodiments 1 to 15, wherein the method comprises systemic and local administration of the tyrosine kinase inhibitor, optionally wherein the method comprises intratumoral administration of the tyrosine kinase inhibitor.

[0399] 17. The method of any one of embodiments 2 to 16, wherein systemic administration of a tyrosine kinase comprises oral administration, intravenous administration, subcutaneous administration, and / or intraperitoneal administration.

[0400] 18. The method of any one of embodiments 1 to 3 and 5 to 17, wherein the proinflammatory agent and the tyrosine kinase inhibitor are administered within 24 hours of each other, optionally wherein the proinflammatory agent and the tyrosine kinase inhibitor are administered within 4 hours of each other.

[0401] 19. The method of any one of embodiments 1 to 3 and 5 to 18, wherein the method comprises intratumoral administration of the proinflammatory agent.

[0402] 20. The method of any one of embodiments 1 to 3 and 5 to 19, wherein the method comprises administering the pro-inflammatory agent to a site different from the site of the cancer to be treated.

[0403] 21. The method of any one of embodiments 1 to 2 and 5 to 18, wherein the pro- inflammatory agent comprises an agent selected from the group consisting of a TLR agonist, a STING activator, a radiation therapy, a PAMP / DAMP activator, a checkpoint inhibitor, a pro-inflammatory cytokine, a chemotherapeutic agent, a bacterial component, a cancer vaccine, an oncolytic virus, a sound therapy, a magnet therapy, an electric therapy, and an electrostatic therapy.

[0404] 22. The method of any one of embodiments 1 to 3 and 5 to 21, wherein the pro- inflammatory agent comprises a TLR agonist.

[0405] 23. The method of embodiment 22, wherein the TLR agonist activates a TLR on a macrophage, optionally wherein the TLR comprises TLR2, TLR3, TLR7, TLR8, and / or TLR9.

[0406] 24. The method of embodiment 23, wherein the TLR agonist comprises CpG, poly I:C, and / or R848.

[0407] 25. The method of any one of embodiments 1 to 3 and 5 to 24, wherein the pro- inflammatory agent comprises a bacterial component, optionally the bacterial component comprises lipopolysaccharide (LPS).

[0408] 26. The method of any one of embodiments 1 to 3 and 5 to 25, wherein the pro- inflammatory agent comprises a STING activator.

[0409] 27. The method of embodiment 26, wherein the STING activator comprises 2'3'-cGAMP.

[0410] 28. The method of any one of embodiments 1 to 3 and 5 to 27, wherein the pro- inflammatory agent comprises a chemotherapeutic agent.

[0411] 29. The method of embodiment 28, wherein the chemotherapeutic agent comprises azathioprine (AZA).

[0412] 30. The method of any one of embodiments 1 to 3 and 5 to 29, wherein the pro- inflammatory agent comprises a pro-inflammatory cytokine.

[0413] 31. The method of embodiment 30, wherein the pro-inflammatory cytokine comprises IL-1b, IL-18, IL-6, and / or TNFa.

[0414] 32. The method of any one of embodiments 1 to 3 and 5 to 31, wherein the proinflammatory agent comprises radiation therapy.

[0415] 33. The method of embodiment 32, wherein the radiation therapy comprises irradiation at a site of the cancer to be treated.

[0416] 34. The method of embodiment 32 or embodiment 33, wherein the radiation therapy comprises irradiation at a site different from a site of the cancer to be treated.

[0417] 35. The method of any one of embodiments 32 to 34, wherein the dose of the radiation therapy is insufficient to kill tumor cells.

[0418] 36. The method of any one of embodiments 1 to 3 and 5 to 35, wherein the proinflammatory agent comprises a checkpoint inhibitor.

[0419] 37. The method of embodiment 36, wherein the checkpoint inhibitor comprises an anti-PD-Ll antibody, an anti-PD-1 antibody, or an anti-CLTA4 antibody.

[0420] 38. The method of any one of embodiments 1 to 3 and 5 to 37, wherein the proinflammatory agent is administered intermittently.

[0421] 39. The method of any one of embodiments 1 to 3 and 5 to 38, wherein the proinflammatory agent and the tyrosine kinase inhibitor are administered simultaneously or concurrently.

[0422] 40. The method of any one of embodiments 1 to 3 and 5 to 39, wherein the proinflammatory agent comprises an immune cell.

[0423] 41. The method of any one of embodiments 4 to 39, wherein the method further comprises administering an immune cell.

[0424] 42. The method of embodiment 40 or 41, wherein the immune cell is derived from the same individual.

[0425] 43. The method of any one of embodiments 40 to 42, wherein the immune cell comprises or is a macrophage, optionally wherein the macrophage has an Ml phenotype.

[0426] 44. The method of any one of embodiments 40 to 43, wherein the immune cell is derived from a monocyte.

[0427] 45. The method of any one of embodiments 40 to 44, wherein the immune cell expresses high levels of MHC-I, MHC-II, CD80, and / or CD86.

[0428] 46. The method of any one of embodiments 40-45, wherein the immune cells express one or more pro-inflammatory cytokines, optionally wherein the one or more pro-inflammatory cytokines comprise TNFa and / or IL-12.

[0429] 47. The method of any one of embodiments 40-46, wherein the immune cells do not express significant levels of TGFp and / or IL-10.

[0430] 48. The method of any one of embodiments 40-47, wherein the immune cells comprise T cells.

[0431] 49. The method of any one of embodiments 40-48, wherein the immune cells are engineered to express a chimeric antigen receptor, optionally wherein the chimeric antigen receptor specifically binds to a tumor antigen.

[0432] 50. The method of any one of embodiments 43-49, wherein the macrophages are engineered to be tyrosine kinase expression and / or activation deficient.

[0433] 51. The method of any one of embodiments 40-50, wherein the tyrosine kinase inhibitor and the immune cells are administered within 24 hours of each other, optionally wherein the tyrosine kinase inhibitor and the immune cells are administered within 4 hours of each other.

[0434] 52. The method of any one of embodiments 40-51, wherein the immune cells are administered concurrently or concurrently with the tyrosine kinase inhibitor.

[0435] 53. The method of any one of embodiments 1-52, further comprising administering to the individual an effective amount of a SHP-1 inhibitor.

[0436] 54. The method of any one of embodiments 1-53, further comprising administering to the individual an effective amount of an anti-TNFa antibody.

[0437] 55. The method of any one of embodiments 1-54, wherein the cancer is a solid tumor.

[0438] 56. The method of any one of embodiments 1-54, wherein the cancer is a hematological cancer.

[0439] 57. The method of any one of embodiments 1-56, wherein the cancer is an advanced cancer.

[0440] 58. The method of any one of embodiments 1-57, wherein the cancer is resistant or refractory to radiation therapy, a chemotherapeutic agent, and / or a checkpoint inhibitor.

[0441] 59. The method as described in any one of embodiments 1 to 58, wherein the individual is a person.

[0442] 60. A composition comprising a tyrosine kinase inhibitor and a pro-inflammatory agent, wherein the pro-inflammatory agent optionally comprises an agent selected from the group consisting of immune cells, TLR agonists, STING activators, agents for use in radiotherapy, PAMP / DAMP activators, checkpoint inhibitors, pro-inflammatory cytokines, chemotherapeutic agents, bacterial components, cancer vaccines, oncolytic viruses, and agents for use in sound therapy, magnetotherapy, electrotherapy, or electrostatic therapy.

[0443] 61. The composition as described in embodiment 60, further comprising an SHP-1 inhibitor.

[0444] Example

[0445] The following embodiments are intended to illustrate the invention only and should not be construed as limiting the invention in any way. The following embodiments and details are provided by way of illustration rather than limitation.

[0446] Example 1. RK-20449 and dasatinib are effective in tumor treatment in tumor-associated macrophage (TAM) activation models and syngeneic mouse tumor models.

[0447] In vitro assays. This experiment tested the effects of RK-20449 and dasatinib on the response of human and mouse macrophages to TLR stimulation in the presence of cancer cells.

[0448] Human monocyte-derived macrophages (M) were pretreated with RK-20449 and dasatinib for 15 min, followed by the addition of human SW620 colorectal cancer cells (2:1 ratio to M) and a mixture of TLR agonists (αTLR: CpG, PolyIC, and R848, 20 μg / ml each). After incubation at 37°C for 15 min, the cells were gently washed to remove most of the SW620 cells, and then lysed using Hank's buffer (pH 7.2) containing 1% Triton and 1 mM PMSF. Protein tyrosine p...

Claims

1. A method of treating cancer in an individual comprising administering to the individual a) a tyrosine kinase inhibitor, and b) a pro-inflammatory agent.

2. The method of claim 1, wherein the method comprises systemically administering the tyrosine kinase inhibitor.

3. The method of claim 1 or 2, wherein the pro-inflammatory agent comprises an agent selected from the group consisting of a TLR agonist, a STING activator, a PAMP / DAMP activator, a chemotherapy, a pro-inflammatory cytokine, a cancer vaccine, a bacterial component, a sound therapy, a magnet therapy, an electric therapy, and an electrostatic therapy.

4. A method of treating cancer in an individual comprising administering to the individual a tyrosine kinase inhibitor, wherein the individual is in an inflammatory response.

5. The method of any one of claims 1 to 4, wherein the method further comprises intermittently administering the tyrosine kinase inhibitor to the individual, optionally wherein a) the tyrosine kinase inhibitor is administered at least three times; the tyrosine kinase inhibitor is administered at least twice at intervals of no more than once every three days; and / or the tyrosine kinase inhibitor is administered to the individual for at least two cycles, wherein each cycle has from about three days to about twenty days.

6. The method of any one of claims 1 to 5, wherein the tyrosine kinase inhibitor is an inhibitor of a tyrosine kinase of the Src family.

7. The method of any one of claims 1 to 6, wherein the tyrosine kinase inhibitor inhibits any one or more of SRC, BLK, HCK, FYN, FGR, and YES.

8. The method of any one of claims 1 to 7, wherein the tyrosine kinase inhibitor is selected from the group consisting of RK-20449, dasatinib, poziotinib, bosutinib, sarcatinib, KX2-391, and R406.

9. The method of any one of claims 1 to 8, wherein the tyrosine kinase inhibitor inhibits SHP-1 signaling.

10. The method of any one of claims 1 to 9, wherein the tyrosine kinase inhibitor is selected from the group consisting of a small molecule, a nucleic acid (e.g., siRNA, shRNA, antisense RNA, microRNA), a nucleic acid editing system (e.g., a CRISPR system), and a protein agent (e.g., an antibody agent targeting a tyrosine kinase or an activated tyrosine kinase).

11. The method of any one of claims 1 to 10, wherein the method comprises systemically and locally administering the tyrosine kinase inhibitor, optionally wherein the method comprises intratumorally administering the tyrosine kinase inhibitor.

12. The method of any one of claims 1 to 3 and 5 to 11, wherein the pro-inflammatory agent and the tyrosine kinase inhibitor are administered within 24 hours of each other, optionally wherein the pro-inflammatory agent and the tyrosine kinase inhibitor are administered within 4 hours of each other, optionally wherein the method comprises administering the pro-inflammatory agent intratumorally, optionally wherein the method comprises administering the pro-inflammatory agent to a different site than the site of the cancer to be treated.

13. The method of any one of claims 1-2 and 5-12, wherein the pro-inflammatory agent comprises an agent selected from the group consisting of a TLR agonist, a STING activator, radiation therapy, a PAMP / DAMP activator, a checkpoint inhibitor, a pro-inflammatory cytokine, a chemotherapeutic agent, a bacterial component, a cancer vaccine, an oncolytic virus, a sound therapy, a magnet therapy, an electric therapy, and an electrostatic therapy.

14. The method of any one of claims 1-3 and 5-13, wherein the pro-inflammatory agent comprises a TLR agonist, optionally wherein the TLR agonist activates a TLR on a macrophage, optionally wherein the TLR comprises TLR2, TLR3, TLR7, TLR8, and / or TLR9, optionally wherein the TLR agonist comprises CpG, poly I:C, and / or R848.

15. The method of any one of claims 1-3 and 5-14, wherein the pro-inflammatory agent comprises a bacterial component, optionally the bacterial component comprises lipopolysaccharide (LPS).

16. The method of any one of claims 1-3 and 5-15, wherein the pro-inflammatory agent comprises a STING activator, optionally wherein the STING activator comprises 2'3'-cGAMP.

17. The method of any one of claims 1-3 and 5-16, wherein the pro-inflammatory agent comprises a chemotherapeutic agent, optionally wherein the chemotherapeutic comprises azathioprine (AZA).

18. The method of any one of claims 1-3 and 5-17, wherein the pro-inflammatory agent comprises a pro-inflammatory cytokine, optionally wherein the pro-inflammatory cytokine comprises IL-1b, IL-18, IL-6, and / or TNFa.

19. The method of any one of claims 1-3 and 5-18, wherein the pro-inflammatory agent comprises radiation therapy, optionally wherein the radiation therapy comprises irradiation at a site of the cancer to be treated, and optionally wherein the radiation therapy comprises irradiation at a site different from the site of the cancer to be treated, optionally wherein the dose of the radiation therapy is insufficient to kill tumor cells.

20. The method of any one of claims 1-3 and 5-19, wherein the pro-inflammatory agent comprises a checkpoint inhibitor, optionally wherein the checkpoint inhibitor comprises an anti-PD-Ll antibody, an anti-PD-1 antibody, or an anti-CLTA4 antibody.

21. The method of any one of claims 1-3 and 5-20, wherein the pro-inflammatory agent and the tyrosine kinase inhibitor are administered simultaneously or concurrently.

22. The method of any one of claims 1-3 and 5-21, wherein the pro-inflammatory agent comprises an immune cell, optionally wherein the immune cell is derived from the same individual.

23. The method of any one of claims 4-22, wherein the method further comprises administering an immune cell, optionally wherein the immune cell is derived from the same individual, optionally wherein: a) the immune cells comprise or are macrophages, optionally wherein the macrophages have an Ml phenotype, optionally wherein the macrophages are engineered to be tyrosine kinase expression and / or activation deficient; b) the immune cells are derived from monocytes; c) the immune cells express high levels of MHC-I, MHC-II, CD80, and / or CD86; d) the immune cells express one or more pro-inflammatory cytokines, optionally wherein the one or more pro-inflammatory cytokines comprise TNFa and / or IL-12; e) the immune cells do not express significant levels of TGF and / or IL-10; f) the immune cells comprise T cells; g) the immune cells are engineered to express a chimeric antigen receptor, optionally wherein the chimeric antigen receptor specifically binds to a tumor antigen; and / or h) the tyrosine kinase inhibitor and the immune cells are administered within 24 hours of each other, optionally wherein the tyrosine kinase inhibitor and the immune cells are administered within 4 hours of each other; and / or the immune cells are administered concurrently or concurrently with the tyrosine kinase inhibitor.

24. The method of any one of claims 1 to 23, further comprising administering to the individual an effective amount of a SHP-1 inhibitor.

25. The method of any one of claims 1 to 24, further comprising administering to the individual an effective amount of an anti-TNFa antibody.

26. The method of any one of claims 1 to 25, wherein the cancer is a solid tumor.

27. The method of any one of claims 1 to 26, wherein the cancer is a hematological cancer.

28. The method of any one of claims 1 to 27, wherein the cancer is an advanced cancer.

29. The method of any one of claims 1 to 28, wherein the cancer is resistant or refractory to radiation therapy, a chemotherapeutic agent, and / or a checkpoint inhibitor.

30. The method of any one of claims 1 to 29, wherein the individual is a human.

31. A composition comprising a tyrosine kinase inhibitor and a pro-inflammatory agent, optionally wherein the pro-inflammatory agent comprises an agent selected from the group consisting of an immune cell, a TLR agonist, a STING activator, an agent for use in radiation therapy, a PAMP / DAMP activator, a checkpoint inhibitor, a pro-inflammatory cytokine, a chemotherapeutic agent, a bacterial component, a cancer vaccine, an oncolytic virus, and an agent for use in sound therapy, magnetotherapy, electrotherapy, or electrostatic therapy.

32. The composition of claim 31, further comprising a SHP-1 inhibitor.

Images