SHP-1 inhibitors for treatment of cancer

Abstract

The present application provides methods of treating cancer in a subject involving administering to the subject an SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a pro-inflammatory agent (e.g., a TLR agonist, radiation therapy, or an immune checkpoint inhibitor). In some cases, the methods involve administering an SHP-1 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 / 404,392, filed September 7, 2022, and U.S. Provisional Application No. 63 / 491,008, filed March 17, 2023, the contents of each of which are hereby incorporated by reference in their entirety. Technical Field

[0003] The present invention relates to compositions and methods for treating cancer involving the administration of a SHP-1 inhibitor and, optionally, a pro-inflammatory agent. Background of the Invention

[0005] In cancers such as solid tumors, intratumoral bone marrow leukocytes, including macrophages (i.e., tumor-associated macrophages or TAMs) and myeloid-derived suppressive cells (MDSCs), play a key role in controlling the immunosuppression of the tumor microenvironment (TME), which supports tumor growth and also confers resistance to immunotherapeutic treatment. An important mechanism by which intratumoral bone marrow leukocytes adapt to an immunosuppressive phenotype and enhance their immunosuppressive capacity following tumor therapy is through multiple negative regulatory pathways mediated by their cell surface inhibitory receptors (iRs). Under tumor therapy, phosphorylation of iRs in the immunoreceptor tyrosine-based inhibitory motif (ITIM) of their cytoplasmic domain leads to activation of the central signaling regulator SHP-1, which mediates dephosphorylation and thus inactivates multiple signaling molecules, resulting in a reduction in the anti-cancer proinflammatory response induced by therapeutic agents ( Figure 1 In solid tumors, as tumors progress to late stages, TME expression of essential cell surface iRs (e.g., SIRPα, Siglec, LilRB, PirB, LAIR1, lectin receptors, SLAM family receptors, etc. (1,2)) also increases, which are regulated by activating SHP-1, which then mediates downstream inhibition.

[0006] Given these inhibitory mechanisms elucidated in the past few years, a pipeline of therapeutic agents aimed at blocking iRs (e.g., anti-LilRB1 / 2 and anti-SIRPα) and their ligands (e.g., anti-CD47) is under development (3-5). However, these efforts, which target each iR or its ligand singly rather than all inhibitory pathways simultaneously, have achieved weak to partial efficacy in controlling solid tumors.

[0007] The disclosures of all publications, patents, patent applications, and published patent applications mentioned herein are hereby incorporated by reference in their entirety. Summary of the Invention

[0008] In one aspect, the present application provides a method of treating cancer in an individual, comprising administering to the individual a) a SHP-1 inhibitor, and b) a proinflammatory agent, wherein the method comprises intermittently administering the SHP-1 inhibitor to the individual. In some embodiments, the method comprises administering the SHP-1 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, radiotherapy, 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, acoustic wave therapy, magnetic therapy, electrical therapy, and electrostatic therapy.

[0009] In another aspect, the present application provides a method of treating cancer in an individual, comprising administering to the individual a) a SHP-1 inhibitor, and b) a proinflammatory agent, wherein the method comprises systemically administering the SHP-1 inhibitor. In some embodiments, the method comprises intermittently administering the SHP-1 inhibitor to the individual. In some embodiments, the proinflammatory 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 proinflammatory cytokine, a chemotherapeutic agent, a bacterial component, a cancer vaccine, an oncolytic virus, acoustic wave therapy, magnetic therapy, electrical therapy, and electrostatic therapy.

[0010] In another aspect, the present application provides a method of treating cancer in an individual, comprising administering to the individual a) a SHP-1 inhibitor and b) a proinflammatory agent, wherein the proinflammatory agent comprises an agent selected from the group consisting of a TLR agonist, a STING activator, a PAMP / DAMP activator, chemotherapy, a proinflammatory cytokine, a cancer vaccine, a bacterial component, acoustic wave therapy, magnetic therapy, electrical therapy, and electrostatic therapy. In some embodiments, the method comprises intermittently administering the SHP-1 inhibitor to the individual. In some embodiments, the method comprises systemically administering the SHP-1 inhibitor.

[0011] In another aspect, the present application provides a method of treating cancer in an individual, comprising administering a SHP-1 inhibitor to the individual, wherein the individual is experiencing an inflammatory response or has a persistent infection. In some embodiments, the method comprises intermittently administering the SHP-1 inhibitor to the individual. In some embodiments, the method comprises systemically administering the SHP-1 inhibitor. In some embodiments, the method further comprises administering immune cells.

[0012] In some embodiments according to any of the methods described above, the method comprises administering the SHP-1 inhibitor to the individual at least twice at intervals no more than once every three days.

[0013] In some embodiments according to any of the methods described above, the method comprises administering the SHP-1 inhibitor to the individual for at least two cycles, wherein each cycle has about three to about twenty days.

[0014] In some embodiments according to any of the methods described above, the half-life of the SHP-1 inhibitor does not exceed about 5 days, optionally the half-life of the SHP-1 inhibitor does not exceed about 3 days.

[0015] In some embodiments according to any of the methods described above, the SHP-1 inhibitor is effective to inhibit greater than 50% of SHP-1 activity for no more than about 5 days, optionally wherein the SHP-1 inhibitor is effective to inhibit greater than 50% of SHP-1 activity for no more than about 3 days.

[0016] In some embodiments of any of the methods described above, the SHP-1 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., antibodies that target or activate SHP-1), or SH2 domain-containing protein agents that inhibit SHP-1 activation by competing for binding to the ITIM motif, or tyrosine kinase inhibitors that inhibit ITIM phosphorylation and thereby inhibit SHP-1 activation. In some embodiments, the SHP-1 inhibitor is selected from the group consisting of TPI-1 or an analog or derivative thereof, a vitamin E derivative, phomoxanthone A (PXA), and a PKCθ activator. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0017] In some embodiments according to any of the methods described above, the SHP-1 inhibitor is administered at least three times.In some embodiments according to any of the methods described above, the method comprises systemic and local administration of the SHP-1 inhibitor, optionally wherein the method comprises intratumoral administration of the SHP-1 inhibitor.

[0018] In some embodiments according to any of the methods described above, systemic administration of SHP-1 includes oral administration, intravenous administration, subcutaneous administration, and / or intraperitoneal administration.

[0019] In some embodiments according to any of the methods described above, the proinflammatory agent and the SHP-1 inhibitor are administered within about 24 hours of each other (eg, within about 16 hours, 8 hours, 4 hours, 2 hours, 1 hour, or 0.5 hours).

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

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

[0022] In some embodiments according to any of the methods described above, the proinflammatory agent includes a TLR agonist. In some embodiments, the TLR agonist activates the TLR on macrophages. In some embodiments, the TLR includes TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8 and / or TLR9. In some embodiments, the TLR agonist includes CpG, polyI:C and / or R848, flagellin (TLR5), zymosan (TLR2 / 4), DAMPs (such as HMGB1 (TLR2 / 4)) produced by radiotherapy, DNA and RNA molecules (TLR3 / 7 / 8 / 9) etc.

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

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

[0025] In some embodiments according to any of the methods described above, the proinflammatory agent comprises a chemotherapeutic agent. In some embodiments, the chemotherapy comprises azathioprine (AZA).

[0026] In some embodiments according to any of the methods described above, the proinflammatory agent comprises a proinflammatory cytokine. In some embodiments, the proinflammatory cytokine comprises an IL-1 family cytokine (e.g., IL-1b, IL-18), IL-6, IL-17, a TNF family cytokine (e.g., TNFα), and a combination thereof with type I and type II interferons (IFNα, IFNβ, and IFNγ).

[0027] In some embodiments of any of the methods described above according to the application, the proinflammatory agent comprises radiation therapy. In some embodiments, the radiation therapy comprises irradiating the site of the cancer to be treated. In some embodiments, the radiation therapy comprises irradiating a site different from the site of the cancer to be treated. In some embodiments, the dose of the radiation therapy is non-ablative and is insufficient to eliminate the tumor (kill all tumor cells).

[0028] In some embodiments according to any of the methods described above, the proinflammatory agent comprises a checkpoint inhibitor. In some embodiments, the checkpoint inhibitor comprises an anti-PD-L1 antibody, an anti-PD-1 antibody, or an anti-CLTA4 antibody.

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

[0030] In some embodiments according to any of the methods described above, the proinflammatory agent and the SHP-1 inhibitor are administered simultaneously or concurrently.

[0031] In some embodiments according to any of the methods described above, the proinflammatory agent comprises an immune cell. In some embodiments, the immune cells are derived from the same individual. In some embodiments, the immune cells comprise or are macrophages, optionally wherein the macrophages have a proinflammatory (M1) phenotype. In some embodiments, the immune cells are derived from monocytes. In some embodiments, the immune cells express high levels of MHC-I, MHC-II, CD80, and / or CD86. In some embodiments, the immune cells express one or more proinflammatory cytokines, optionally wherein the one or more proinflammatory cytokines comprise TNFα and / or IL-12. In some embodiments, the immune cells do not express significant levels of TGFβ and / or IL-10. In some embodiments, the immune cells comprise T cells. In some embodiments, the immune cells are engineered to express a chimeric antigen receptor, optionally wherein the chimeric antigen receptor specifically binds to a tumor antigen. In some embodiments, the macrophages are engineered to have defective SHP-1 expression and / or activation. In some embodiments, the SHP-1 inhibitor and the immune cells are administered within 24 hours of each other, optionally wherein the SHP-1 inhibitor and the immune cells are administered within 4 hours of each other. In some embodiments, the immune cells are administered simultaneously or concurrently with the SHP-1 inhibitor.

[0032] In some embodiments according to any of the methods described above, 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-TNFα antibodies and anti-IL6 antibodies. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm is administered simultaneously with a tyrosine kinase inhibitor. In some embodiments, the agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm is administered sequentially with a tyrosine kinase inhibitor (e.g., before or after it). 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 schedule as the tyrosine kinase inhibitor.

[0033] In some embodiments according to any of the methods described above, the cancer is a solid tumor.

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

[0035] In some embodiments according to any of the methods described above, the cancer is an advanced cancer.

[0036] In some embodiments according to any of the methods described above, the cancer is resistant or refractory to radiation therapy, chemotherapeutic agents, and / or checkpoint inhibitors.

[0037] In some embodiments according to any of the methods described above, the individual is a human.

[0038] In another aspect, the present application provides a composition comprising a SHP-1 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 radiotherapy, 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 acoustic wave therapy, magnetic therapy, electrical therapy, or electrostatic therapy. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 Depicted, SHP-1 acts as a "master" signaling mediator downstream of multiple inhibitory receptors on myeloid leukocytes in the tumor microenvironment (TME). SHP-1 activity attenuates RT-induced and immunotherapy-induced pro-inflammatory pathways and anti-cancer efficacy, and maintains the myeloid leukocyte immunosuppressive phenotype. Our SHP-1 inhibition approach as an anti-cancer strategy (red). Partially listed are companies and approaches aimed at depleting or blocking individual cell surface inhibitory receptors, including SIRPα (SIRPαnt Immunotherapeutics, anti-CD47 Gilead) and anti-SIRPα (Biosion) approaches), Siglec (NextCure), LilRB (Next-IO), SLAMF (BMS), etc.

[0040] Figures 2A to 2G Depicted, inhibition of SHP-1 enhances macrophage proinflammatory responses, antigen presentation, and phagocytosis in the tumor environment. Figure 2A and Figure 2B depicts that SHP-1 activity in macrophages is mediated by proinflammatory stimuli and extracellular cancer cell attachment ("contact") of macrophages ( Figure 2A The activity of SHP-1 was inhibited in a dose-dependent manner by the catalytic domain covalent inhibitor TPI-1 (1 μg / ml, Figure 2A )inhibition( Figure 2B The SHP-2 inhibitor SHP099 (1 μg / ml) was used in the assay. Figure 2C It is depicted that TPI-1 inhibition of SHP-1 dose-dependently restored IFNγ / LPS-induced activation of STAT1 (p-STAT1) and Erk1 / 2 (p-Erk1 / 2) inactivated by SHP-1 (red lines). Figure 2D and Figure 2E depicts that inhibition of SHP-1 by TPI-1 in macrophages enhances the production of pro-inflammatory cytokines ( Figure 2D ) and the expression of immunogenic antigen presentation mechanisms via IFNγ / LPS in the tumor environment ( Figure 2E ). Figure 2F and Figure 2G Depicts that SHP-1 inhibition promotes phagocytosis of cancer cells by pro-inflammatory activated macrophages. Figure 2F Human monocyte-derived macrophages were shown to phagocytose THP-1 leukemia cells and HT29 colon cancer cells. Figure 2G Murine bone marrow-derived macrophages are shown phagocytosing isogenic cancer cells EL4 (T lymphoma), B16 (melanoma), MC38 (colorectal cancer), Pan01 and KPC (both pancreatic adenocarcinomas), and LLC (lung cancer). The SHP-1 and SHP-2 inhibitors TPI-1 and SHP099 were used in the assay at 1 μg / ml, respectively.

[0041] Figures 3A to 3E To characterize the regulation of SHP-1 activity in proinflammatory macrophages in cancer. Macrophages co-cultured with cancer cells were stimulated with IFNγ / LPS or a TLR agonist (αTLR) containing CpG, polycytidylic acid (i.e., polyI:C), and R848 (0.4 μg / ml each) for 30 min (37°C) in the presence or absence of various SHP-1 or SHP-2 inhibitors or activators, followed by cell lysis and PTP activity assays. Figure 3A Delineate the regulatory mechanism of SHP-1 activity. Figure 3B and Figure 3C Depicts the presence of cancer cell junctions mediated by IFNγ / LPS ( Figure 3B ) or TLR agonists ( Figure 3C )-induced PTP activity in macrophages was largely attenuated (>70%) by inhibition of SHP-1 (TPI-1 and PTP-I) or the pan-PTP inhibitors PTP-III and pervanadate, but was only weakly reduced (<10%) by inhibition of SHP-2 (SHP099 and PHPS1). Figure 3D It is depicted that vitamin E derivatives and Phomopsis xanthone A (PXA) dose-dependently and appropriately inhibited SHP-1 activity in IFNγ / LPS-stimulated macrophages surrounding cancer cells. Figure 3E Figure 3: PKCθ negatively regulates SHP-1 activity at a modest level. Inhibition of PKCθ by PKCθ inhibitors I and VTX27 increased SHP-1 activity through IFNγ / LPS and cancer cell ligation. Conversely, PKCθ activation by PMA decreased SHP-1 activity.

[0042] Figure 4A and Figure 4B Depicted, pulsed inhibition of SHP-1 transiently enhances pro-inflammatory signaling in macrophages. Figure 4A Murine macrophages (bone marrow-derived macrophages or "BMDM") were treated with TPI-1 for 15 minutes and then washed to completely remove TPI-1. At various time points after TPI-1 treatment, macrophages were stimulated with IFNγ / LPS for 20 minutes in the presence of live cancer cells (1:1 ratio to BMDM), after which the cells were lysed and SHP-1 activity was measured using pNpp and total and phosphorylated STAT1 and Erk1 / 2 were detected by Western blotting. Figure 4B :and Figure 4A The same setup was used as in the experiment, except that after TPI-1 treatment, TPI-1 was partially depleted (50%) or not in the culture medium, followed by macrophage stimulation with IFNγ / LPS at different time points.

[0043] Figures 5A to 5C Delineating signaling mechanisms in solid tumors when targeting SHP-1. Figure 5A Describe an overview of SHP-1 mechanisms in solid tumors. SHP-1 remains inactive / low activity in solid tumors until therapeutic treatment. SHP-1 is activated following the following tumor-protective feedback loop: proinflammatory signaling → tyrosine kinase (TK) → ITIM phosphorylation → SHP-1 activation. SHP-1 activity then inhibits proinflammatory signaling and confers tumor resistance to therapy. TPI-1 inhibits SHP-1 activity and restores proinflammatory signaling, leading to anti-tumor innate and adaptive immunity. Figure 5B An example of this is depicted in SIRPα. Proinflammatory signals induce one or more Src family TKs to mediate docking of SHP-1, leading to phosphorylation in the ITIM, the cytoplasmic domain of SHP-1 activation. Figure 5C Figure 3: Proinflammatory stimuli-induced phosphorylation of the iR ITIM leads to exclusive binding of SHP-1 to the pITIM, resulting in SHP-1 activation. Immunosuppressive signaling through IL-4, IL-10, and TGFβ induces ITIM phosphorylation, leading to SHP-1 binding. Examples show macrophage iRs LilRB and SIRPα.

[0044] 6A to 6D It is depicted that, in different contexts, inhibition of SHP-1 promotes either anti-tumor or pro-tumor effects. Figure 6A Figure 3 shows that intratumoral macrophages upregulate the expression of iRs (Pir-B; Siglec E, F, and G; and SIRPα) and that tumors progress to late, larger sizes. 3 ) of KPC pancreatic tumors and growth to larger size (>800mm 3) were dissociated into single cells, followed by flow cytometry analysis of cell surface protein expression on macrophages (gated F4 / 80+). Figure 6B As depicted, inhibition of SHP-1 alone promotes TME immunosuppression. Excised MC38 solid tumor cubes were treated with the SHP-1 inhibitor TPI-1 (100 nM) or vehicle (DMSO) in a cell culture environment. After 24 hours, cytokine secretion into the tumor in culture was measured by ELISA. As shown, TPI-1 treatment increased tumor production of IL-6 and IL-10. Figure 6C Figure 3: Inhibition of SHP-1 by TPI-1 and PTP-1 dose-dependently enhances IL-10 and TGFβ production by macrophages in the presence of cancer cell junctions under immunosuppressive stimulation by IL-4 / 13 or IL-10. Figure 6D TPI-1 and PTP-1 dose-dependently enhanced macrophage proinflammatory responses, as evidenced by increased IL-12 and TNFα induced by IFNγ and LPS.

[0045] 7A to 7F Depict that inhibition of SHP-1 unleashes proinflammatory responses and antigen presentation in solid tumors following therapy. Figure 7A Intratumoral treatment of KPC pancreatic tumors with TLR agonists (αTLR containing CpG, polyI:C, and R848, 1 μg each), proinflammatory cytokines (IL-1β, IL-6, TNFα, and IFNγ, 10 ng each), and the STING activator 2'3'-cGAMP (1 μg) for 30 min induced a spike in PTP activity, which was attenuated by the SHP-1 inhibitor TPI-1. Low PTP / SHP-1 activity was found in untreated steady-state tumors. Figure 7B showed that similarly, PTP / SHP-1 activities in KPC tumors were induced by one fraction of 8 Gy RT treatment, one dose of chemotherapy with azathioprine (AZA) or anti-PD-L1 antibody (αPD-L1), and these activities were attenuated by simultaneous treatment with TPI-1. Figure 7C Depicts that depletion of intratumoral macrophages with clodronate liposomes abrogates SHP-1 activity induced by various tumor treatments. Figure 7D and Figure 7E showed that inhibition of SHP-1 largely enhanced the expression of proinflammatory cytokines ( Figure 7D ) and the immunogenic antigen presentation capacity of intratumoral macrophages ( Figure 7E ). Of note, treatment of tumors with TLR agonists or RT in the absence of SHP-1 inhibition induced minimal increases in proinflammatory cytokines, but significant increases in IL-10 and TGFβ. Figure 7FDepicted, transcriptional profiles revealed distinct tumor responses to TLR agonists and RT without and with intratumoral SHP-1 inhibition. Without SHP-1 inhibition, KPC tumors exhibited treatment resistance with increased immunosuppressive TGFβ signaling and MDSC infiltration, whereas treated tumors with SHP-1 inhibition reprogrammed the TME into a potent pro-inflammatory niche with reduced TGFβ but high expression of inflammatory cytokines, antigen-presenting molecules, and chemokines that attract neutrophils, NK and T cells, but not MDSCs. Similar data were obtained by studying colorectal carcinoma MC38.

[0046] Figures 8A to 8E Depicted, SHP-1 inhibition in combination with a TLR agonist (αTLR) reprograms the TME of MC38 colorectal cancer. Figure 8A Depicts the MC38 tumor treatment protocol. Figures 8B to 8C The results showed that TME analysis confirmed that TME was reprogrammed by combined treatment with TPI-1 and αTLR, inducing tumor cell reduction and immune infiltration, especially an increase in tumor-destructive CD8 T cells, neutrophils (PMNs) and NK cells, while reducing macrophages, MDSCs and Tregs. Figure 8D It showed that after TPI-1 and αTLR treatment, the number of intratumoral macrophages was significantly reduced. Figure 8E It was shown that ex vivo treatment of resected MC38 tumors with TPI-1 and αTLR induced CD8 T cell expansion, indicating that the treatment induced antigen presentation in situ.

[0047] Figures 9A to 9C Depicted, inhibition of SHP-1 by TPI-1 combined with tumor-localized RT reprograms the TME of KPC pancreatic ductal adenocarcinoma toward proinflammatory cancer elimination. Figure 9A Depicts treatment regimens and TME analysis at day 5. Table depicts the percentage of various populations within total CD45+ cells. Bar graph depicts the percentage of various populations within total cells. Figure 9A It was demonstrated that TPI-1 combined with RT induced neutrophil (PMN) infiltration, NK cell increase, and CD8 T cell expansion. Figure 9B As depicted, TPI-1 combined with RT treatment induced a significant expansion of CD8 T cells with a remarkably high frequency of responses to the tumor-specific antigen p15E. Figure 9C Depicted, intratumoral macrophages exhibit a proinflammatory phenotype and increased antigen-presenting capacity following combined TPI-1 and RT treatment.

[0048] FIG. 10A to FIG. 10B To characterize a pulsed intermittent SHP-1 inhibition (iShp-1) strategy for the treatment of metastatic solid tumors. Figure 10AA preclinical metastatic solid tumor model was established in syngeneic WT mice via multisite transplantation. After tumor formation, mice were treated with a SHP-1 inhibitor in combination with a pro-inflammatory modality to initiate anti-cancer immunity. Treatment was administered either intraperitoneally or subcutaneously for systemic effect. Treatment could also be administered via intratumoral injection (IT). Figure 10B : Treatment regimens and assessments: The pulse-intermittent regimen administered the SHP-1 inhibitor once at the beginning of each cycle, or two or three times continuously (pulse-1, -2, or -3), followed by a rest period (variable days from 2 to 9 days) before the next treatment cycle. Combination modalities (not limited to the list) were administered simultaneously (e.g., TLR agonists, as shown in the accompanying figures) or otherwise administered after a specific dosing schedule. Tumor control efficacy, side effects, and toxicity were evaluated throughout the experiment. Changes in TME immunogenicity were also determined for mechanistic insights.

[0049] Figures 11A to 11E The effects of continuous or intermittent iShp-1 treatment on efficacy and adverse toxicities were characterized. Figure 11A Study design is described. Mice bearing KPC pancreatic cancer were treated with TPI-1 (1x daily) continuously or intermittently with a rest day between treatments (intermittent). Three doses of 1, 3, and 10 mg / kg (ip) were tested, and TLR agonists (CpG plus polyI:C, 10 μg each, ip, 1x every 3 days) were administered to initiate an inflammatory response. Figure 11B and Figure 11C Demonstrate tumor treatment efficacy. Tumor imaging ( Figure 11B ) and volume change records ( Figure 11C ) indicate that iShp-1 has similar efficacy in continuous or intermittent regimens. Figure 11D and Figure 11E Side effects were shown. Daily recordings of body weight, blood hemoglobin, proteinuria, and serum alanine aminotransferase (ALT), as well as analysis of splenomegaly at the final point (d11), confirmed a high risk of continuous iSHP-1, which caused anemia, renal injury, splenomegaly, and lung inflammation (not shown). However, intermittent iSHP-1 demonstrated a low risk of side effects.

[0050] 12A to 12D Figure 3: Pulse-intermittent SHP-1 inhibition (iSHP-1) in combination with TLR agonists (αTLR) and / or anti-PD-L1 checkpoint inhibitors effectively treats multifocal MC38 colorectal cancer. Figure 12A Treatment schedule depicted. Mice bearing bilateral MC38 tumors were treated with TPI-1 (sc) and various combinations (via ip or sc) for two days. A five-day rest period was given before a second cycle of treatment was performed on mice bearing residual tumors. Figure 12B and Figure 12CDescribe the changes in tumor volume ( Figure 12B ) and TME reprogramming measure tumor control efficacy, indicating an increase in tumor-killing immune populations while reducing immunosuppression. Figure 12D Acute adverse toxicities as measured by body weight, proteinuria, serum ALT levels, and splenomegaly are depicted.

[0051] 13A to 13E Depicts pulsed-intermittent SHP-1 inhibition (iSHP-1) in combination with RT and αPD-L1 for the treatment of pancreatic and lung cancer. Figure 13A The experimental protocol is outlined. On day 1, mice bearing bilateral KPC pancreatic cancer or LLC lung cancer were treated with a single pulse dose of TPI-1 (3 mg / kg) via ip to systemically inhibit SHP-1 (iSHP-1). Concurrently, tumors in the right flank were treated with 8 Gy of X-ray radiation (RT). After a two-day rest period, on day 4, mice were treated with a second cycle of TPI-1 (ip) at the same dose with RT reduced to 4 Gy for the right flank tumors. Cycle 3 (day 7) iSHP-1 was combined with RT reduced to 2 Gy. Anti-PD-L1 Ab (100 μg, ip) was administered one day after TPI-1 plus RT. Figure 13B Depicted are luminescence images tracking changes in KPC-luc and LLC-luc tumors following treatment. Figure 13C Changes in tumor volume are depicted and animal survival rates were recorded up to 45 days after treatment. Figure 13D and Figure 13E Treatment did not cause splenomegaly, weight loss, or anemia ( Figure 13D ), which also does not cause lung inflammation ( Figure 13E ).

[0052] 14A to 14E Depicted, pulsed-intermittent inhibition of SHP-1 (iSHP-1) combined with a TLR agonist (αTLR) treats advanced KPC pancreatic ductal adenocarcinoma. Figure 14A Treatment schedule is depicted. Mice bearing large KPC tumors were treated (ip) with TPI-I plus TLR agonists (50 μg each) for three consecutive days. After the initial pulse treatment, a 9-day rest period was given before a second two-day treatment cycle with TPI plus TLR agonists. Figure 14B Depicted are luminescent images of KPC tumor-bearing mice during the course of treatment. Figure 14C Depict tumor volume changes. Figure 14D Figure 4. TME analysis at day 4 reveals immunosuppressive populations within the tumor, including macrophages. and MDSCs) decreased, and tumor-killing CD8 T cells (Tc), inflammatory neutrophils (PMNs), and NK cells increased. Figure 14EAs depicted, the treatment regimen caused mild side effects and temporary weight loss, followed by recovery.

[0053] Figure 15 Proteomic analysis characterizing protein tyrosine phosphatase expression in macrophages.

[0054] 16A to 16D Delineating the activation of macrophage proinflammatory responses and antigen presentation in the tumor environment by a combinatorial approach of SHP-1 inhibition (iShp1). Figure 16A Describe the test system. Figure 16B and Figure 16C The effect of interferon with and without TPI-1 is depicted. Figure 16D The effects of various agents, including IL-1 family cytokines (IL-1β, IL-18), TNFα, and TLR ligands, in combination with TPI-1 were profiled.

[0055] 17A to 17C Depicted, inhibition of SHP-1 (iShp1) abrogates tumor-imposed immunosuppression under proinflammatory challenge. Figure 17A Depicted are neutrophil infiltration in different organs measured at different time points after αTLR stimulation. Figure 17B Depicts neutrophil infiltration of tumor tissue in mice treated with αTLR plus TPI-1. Figure 17C Characterizing the phenotype of intratumoral macrophages.

[0056] Figures 18A to 18G Anti-TNFα mAb was shown to curb systemic inflammation and reduce adverse toxicities. Figure 18A Experimental design is shown. In the absence or presence of additional treatment with anti-TNFαmAb or anti-IL-6mAb (150 μg, ip), mice with established MC38 colorectal cancer (200-400 mm 3 ) were treated with αTLR, TPI-1 and dasatinib (Dasatinib) (sc). The treatment was repeated once (d1 and d2). Tumor volume changes were recorded, and the immune infiltration of the tumor TME was analyzed on day 6 after treatment. Figure 18B Shown are the changes in tumor volume after various treatments. Figure 18C and Figure 18D Figure 3 shows the results of TME analysis. Treatment with anti-TNFα mAb or anti-IL-6 mAb did not affect the increase in CD8 T cells (Tc) and NK cells and the decrease in macrophages and MDSCs induced by αTLR / TPI-1 / dasatinib therapy in the TME. Figure 18ETreatment of mice with anti-TNFα mAb, but not anti-IL-6 mAb, showed that the induction of inflammatory cytokines (TNFα, IL-6, IL-1β, IL-10, IFNα, and IFNγ) associated with αTLR / TPI-1 / dasatinib combination therapy was largely reduced. Figure 18F Anti-TNFα treatment also significantly reduced circulating monocyte and PMN chemokines CCL2, CCL5, and CXCL1, while not reducing CXCL10, which is essential for T cell trafficking. Figure 18G showed that anti-TNFα treatment protected mice from splenomegaly and intestinal inflammation commonly associated with αTLR / TPI-1 / dasatinib therapy.

[0057] 19A to 19C and Figure 20 This suggests a mechanism by which tumor cells suppress the proinflammatory response of macrophages in the TME.

[0058] Figures 21A to 21E It was shown that iRs and their ligands are upregulated when tumors progress to late stages.

[0059] Figures 22A to 22E We show that factors produced by cancer cells and by the tumor TME (secretome) induce increased macrophage expression of iRs.

[0060] FIG. 23A to FIG. 23B We show that inhibition of SHP-1 unleashes pro-inflammatory responses in the KPC tumor TME.

[0061] Figure 24 We show that treatment of MC38 tumors with a TLR agonist (αTLR) plus SHP-1 inhibition induces proinflammatory polarization of the TME.

[0062] Figure 25A and Figure 25B The iR→SHP-1 inhibitory axis is shown. Figure 25A Western blot analysis showing activation of JAK-STAT, NFκB, MAPK, and PI3K-Akt signaling pathways and protein phosphorylation triggered by LPS (1 μg / ml) plus IFNγ (40 ng / ml) stimulation. Figure 25B Densitometry analysis showing protein phosphorylation and therefore signal transduction activation.

[0063] Figures 26A to 26C It was shown that Shp1 - / - Macrophages resist the suppression imposed by cancer cells and unleash a pro-inflammatory response.

[0064] Figures 27A to 27C We show that cell surface blockade of iRs or ligands serves as an alternative strategy to deplete the inhibitory iR→SHP-1 axis. DETAILED DESCRIPTION

[0065] In one aspect, the present application provides a method of treating cancer in an individual, the method comprising administering a SHP-1 inhibitor to the individual, wherein the individual a) has received, is receiving, or will receive a pro-inflammatory agent, or b) is in an inflammatory response or has a persistent infection. In another aspect, the present application provides a method of treating cancer in an individual, the method comprising administering to the individual monocytes or macrophages that are defective in SHP-1 expression or activation, and wherein the individual a) has received, is receiving, or will receive a pro-inflammatory agent, or b) is in an inflammatory response or has a persistent infection. In some embodiments, the SHP-1 inhibitor is administered systemically. In some embodiments, the method comprises administering the SHP-1 inhibitor to the individual at least twice at intervals no more than once every three days. In some embodiments, the method comprises administering the SHP-1 inhibitor to the individual for at least two cycles, wherein the SHP-1 inhibitor is administered at least once in each cycle and wherein each cycle has about three to about twenty days. In some embodiments, 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, radiation therapy, acoustic wave therapy, magnetic therapy, electrical therapy, and electrostatic therapy.

[0066] The present application is based, at least in part, on the surprising discovery that combining the "master" inhibitory enforcer SHP-1 with pro-inflammatory therapies unleashes pro-inflammatory signaling in the tumor environment, acting particularly on tumor-infiltrating macrophages, leading to a potent reprogramming of the TME and enhanced activation of innate and adaptive immune cells to promote anti-cancer immunity. Specifically, it has been found that iR-SHP-1-mediated inhibitory regulation within tumors is particularly potent under tumor therapies because these therapies often induce hyperphosphorylation of ITIMs, promoting "hyperactivation" of SHP-1 (a feedback loop that protects tumors from therapeutic injury and inflammatory insults (ahead)), and also elicits a wound healing response to promote tumor progression. See, e.g., Figure 1 and Figure 5A This finding emphasizes the necessity of inhibiting SHP-1 as a combinatorial modality in tumor immunotherapy to achieve efficacy.

[0067] It has been demonstrated that combining SHP-1 inhibitors (e.g., TPI-1) with proinflammatory agents (e.g., TLR agonists, proinflammatory cytokines, radiation therapy, checkpoint inhibitors) achieves the surprising effect of converting the immunosuppressive TME into an inflammatory TME, thereby activating multiple types of immune cells (e.g., macrophages, T cells, and B cells) and completely depleting the tumor. See, e.g., Figure 11C and Figure 12B It has also been demonstrated that this combination therapy achieves remote effects (e.g. Figure 13B ) and it is effective in treating advanced large-sized tumors (e.g. Figure 14C ).

[0068] Furthermore, it has been found that when administered in intermittent dosing, SHP-1 inhibitors are able to achieve significant anti-tumor effects comparable to those of SHP-1 inhibitors administered in continuous dosing, while achieving significantly fewer side effects (e.g., anemia, renal impairment, and liver impairment). 11A to 11D This is surprising given the severe side effects associated with SHP-1 inhibitors demonstrated in previous studies. Furthermore, administration of agents that reduce systemic inflammation (e.g., anti-TNFα mAbs) further inhibits systemic inflammation and reduces adverse toxicities. Figures 18A to 18G .

[0069] Thus, the present application provides novel approaches that can effectively rewire the immunosuppression imposed by tumor disease and allow both innate and adaptive immunity against cancer, thereby achieving significant anti-tumor efficacy.

[0070] I. Definition

[0071] Generally, the terms used in the claims and this specification are intended to be interpreted as having their ordinary meanings as understood by one of ordinary skill in the art. For greater clarity, certain terms are defined below. In the event of a conflict between the ordinary meanings and the provided definitions, the provided definitions will apply.

[0072] The terms "subject," "subject," or "patient" are used synonymously herein to describe mammals, including humans. Subjects include, but are not limited to, humans, cows, horses, cats, dogs, rodents, or primates. In some embodiments, the subject is human. In some embodiments, the subject has a disease, such as cancer. In some embodiments, the subject is in need of treatment.

[0073] As used herein, "reference" refers to any sample, standard, or level used for comparison purposes. A reference can be obtained from a healthy and / or disease-free sample. In some instances, a reference can be obtained from an untreated sample. In some instances, a reference is obtained from a disease-free or untreated sample of an individual. In some embodiments, a reference is obtained from one or more healthy individuals who are not individuals.

[0074] As used herein, the term "intermittent" or "intermittently" in the context of administration refers to non-continuous administration, e.g. Figure 11A (Picture below), Figure 12A 、 Figure 13A and Figure 14AIn some cases, "intermittent" administration refers to administration wherein a) the SHP-1 inhibitor is administered on fewer than 12 consecutive days (e.g., fewer than 11, 10, 9, 8, 7, 6, 5, 4, and 3 days), and b) the SHP-1 inhibitor is administered at least twice, with the two administrations separated by at least one day (i.e., Day 1 and Day 3). In some embodiments, the SHP-1 inhibitor is administered at least twice daily on no more than three consecutive days separated by at least one day.

[0075] As used herein, the term "cycle" in the context of administration refers to a period of time during which there is at least one administration of a SHP-1 inhibitor. Day 1 of a cycle is defined as the day when the first administration of a SHP-1 inhibitor occurs during the period. When there are several consecutive daily administrations of a SHP-1 inhibitor, Day 1 of a cycle is defined as the day when the first of the several consecutive daily administrations occurs. The last day of a cycle is defined as the day before the next non-consecutive administration of a SHP-1 inhibitor occurs. For exemplary cycles, see Figure 12A and Figure 14A The cycles need not be of the same length. For example, a first cycle may have five days, and a second cycle may have seven days. Each cycle may have a different number of SHP-1 inhibitor administrations. For example, a first cycle (which may have five days) may have one SHP-1 inhibitor administration, and a second cycle (which may have seven days) may have two SHP-1 inhibitor administrations.

[0076] As used herein, the term "immunogenicity" is the ability to elicit an immune response, for example, via T cells, B cells, or both.

[0077] As used herein, "treatment" or "treating" is a method for obtaining a beneficial or desired result (including a clinical result). For the purposes of the present invention, a beneficial or desired clinical result includes, but is not limited to, one or more of the following: alleviating one or more symptoms caused by the disease, reducing the extent of the disease, stabilizing the disease (e.g., preventing or delaying worsening of the disease), preventing or delaying the spread of the disease (e.g., metastasis), preventing or delaying the occurrence or recurrence of the disease, delaying or slowing the progression of the disease, improving the condition of the disease, providing relief (whether partial or total relief), reducing the dose of one or more other drugs required to treat the disease, delaying the progression of the disease, improving the quality of life, and / or prolonging survival. "Treatment" also encompasses alleviating cancer pathology results. The methods of the present invention encompass any one or more of these therapeutic aspects.

[0078] As used herein, "delaying" the development of cancer means postponing, hindering, slowing down, blocking, stabilizing and / or postponing the development of the disease. Such delays may have different lengths of time, depending on the history of the disease and / or the individual being treated. It will be apparent to those skilled in the art that sufficient or significant delays may actually encompass prevention, such that the individual does not develop the disease. A method of "delaying" the development of cancer is a method that reduces the probability of disease development and / or alleviates the extent of the disease within a given timeframe, compared to when the method is not used. Such comparisons are typically based on clinical studies conducted using a statistically significant number 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 tests, arterial photography, or biopsy. Development may also refer to cancer progression that may be initially undetectable and includes occurrence, recurrence, and onset.

[0079] As used herein, the term "simultaneous administration" means that the first and second therapies in the combination therapy are administered with a time interval of no more than about 15 minutes (e.g., no more than about 10 minutes, 5 minutes, or 1 minute). When the first and second therapies are administered simultaneously, the first and second therapies can be contained in the same composition (e.g., a composition comprising both the first and second therapies) or separate compositions (e.g., the first therapy is in one composition and the second therapy is in another composition).

[0080] As used herein, the term "sequential administration" means that the first and second therapies in the combination therapy are administered with a time interval of more than about 15 minutes (e.g., more than about any of 20 minutes, 30 minutes, 40 minutes, 50 minutes, 60 minutes, or longer). Either the first or the second therapy can be administered first. The first and second therapies are contained in separate compositions, which can be contained in the same or different packages or kits.

[0081] As used herein, the term "concurrent administration" means that the administration of a first therapy and the administration of a second therapy in the combination therapy overlap with each other.

[0082] As used herein, by "pharmaceutically acceptable" or "pharmacologically compatible" is meant to be free of biologically or otherwise undesirable materials, e.g., the material can be incorporated into a pharmaceutical composition administered to a subject without causing any significant undesirable biological effects or interacting in a deleterious manner with any other component 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 Inactive Ingredient Guide established by the US Food and Drug Administration.

[0083] It should be understood that the embodiments of the present application described herein include "consisting of" embodiments and / or "consisting essentially of" embodiments.

[0084] Reference herein to "about" a value or parameter includes (and describes) variations with respect to the value or parameter itself. For example, a description referring to "about X" includes a description of "X."

[0085] As used herein, reference to "other than" a value or parameter generally means and describes "other than" a value or parameter. For example, a method not for treating type X cancer means that the method is for treating cancer types other than type X.

[0086] As used herein, the term "about XY" has the same meaning as "about X to about Y."

[0087] It must be noted that, as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0088] Any term not directly defined herein should be understood to have a meaning generally associated with that understood in the technical field of the present invention. Certain terms are discussed herein to provide practitioners with additional guidance on the compositions, devices, methods, etc., describing aspects of the present invention and how to make or use them. It should be understood that the same thing can be expressed in more than one way. Therefore, alternative wordings and synonyms can be used for any one or more of the terms discussed herein. It will not be important whether a term is described or discussed in detail in this article. Some synonyms are provided or alternative methods, materials, etc. are provided. Unless explicitly stated, the description of one or more synonyms or equivalents does not exclude the use of other synonyms or equivalents. The use of examples, including term examples, is for illustrative purposes only and does not limit the scope and meaning of aspects of the present invention in this article.

[0089] II. Treatment Methods

[0090] In one aspect, the present application provides methods for treating cancer by administering a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof). The SHP-1 inhibitors (e.g., TPI-1 or an analog or derivative thereof) described herein include any agent comprising a SHP-1 inhibitor portion (e.g., an agent comprising a TPI-1 portion or a derivative or analog thereof). In some embodiments, the SHP-1 inhibitor comprises TPI-1. In some embodiments, the individual being treated has received, is currently receiving, or will receive a pro-inflammatory agent, such as any of the pro-inflammatory agents described herein. In some embodiments, the individual is in the midst of an inflammatory response or has an ongoing infection.

[0091] In some embodiments, the methods comprise administering both a SHP-1 inhibitor and a proinflammatory agent to an individual. In some embodiments, the SHP-1 inhibitor is administered intermittently. In some embodiments, the SHP-1 inhibitor is administered daily for no more than three or two consecutive days and optionally at least twice (which are separated by at least one day). In some embodiments, the SHP-1 inhibitor is administered at least three, four, or five times. In some embodiments, at least two administrations of the SHP-1 inhibitor are separated by two, three, four, five, six, seven, eight, nine, or ten days. In some embodiments, each administration of the SHP-1 inhibitor is separated by at least one day from the previous or next administration of the SHP-1 inhibitor. In some embodiments, the methods comprise systemic administration of the SHP-1 inhibitor.

[0092] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, comprising administering to the individual a) a pro-inflammatory agent (e.g., a TLR agonist, e.g., R848, e.g., radiation therapy), or b) an inflammatory response or persistent infection, and wherein the SHP-1 inhibitor is administered systemically (e.g., intravenously or subcutaneously). In some embodiments, the SHP-1 inhibitor is administered intermittently. In some embodiments, the SHP-1 inhibitor is administered daily for no more than three or two consecutive days and optionally at least twice (separated by at least one day). In some embodiments, the SHP-1 inhibitor is administered at least three, four, or five times. In some embodiments, at least two administrations of the SHP-1 inhibitor are separated by two, three, four, five, six, seven, eight, nine, or ten days. In some embodiments, each administration of the SHP-1 inhibitor is separated by at least one day from the previous or next administration of the SHP-1 inhibitor. In some embodiments, the SHP-1 inhibitor is administered no more than once every two days. In some embodiments, the SHP-1 inhibitor is administered no less than twice and no more than five times within ten consecutive 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 SHP-1 inhibitor is administered simultaneously with the proinflammatory agent. In some embodiments, the SHP-1 inhibitor is administered concurrently with the proinflammatory agent. In some embodiments, the SHP-1 inhibitor and the proinflammatory agent are administered sequentially and within two weeks (e.g., within 10 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or within the same day). In some embodiments, the half-life of the SHP-1 inhibitor is 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 SHP-1 inhibitor is effective to inhibit greater than 50% of SHP-1 activity for no more than about 7 days (e.g., about 5 days, 4 days, or 3 days). In some embodiments, the SHP-1 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 SHP-1 or activate SHP-1). In some embodiments, the SHP-1 inhibitor is selected from the group consisting of: TPI-1 or an analog or derivative thereof, a vitamin E derivative, Phomopsis xanthone A (PXA), and a PKCθ activator. In some embodiments, the method further comprises administering a pro-inflammatory agent locally (e.g., intratumorally) to the individual. 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-TNFα antibody or an anti-IL6 antibody).In some embodiments, the method further comprises administering an anti-TNFα antibody to the individual, optionally wherein the anti-TNFα antibody is administered prior to (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours), concurrently or simultaneously with, or immediately after (within 3, 2, 1, or 0.5 hours) administration of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and / or a proinflammatory agent. In some embodiments, the proinflammatory agent comprises or is selected from the group consisting of: R848, 3M-852A, Motolimod, Bropirimine, and Vesatolimod. 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 SHP-1 inhibitor comprises TPI-1.

[0093] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, the method comprising administering to the individual a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a proinflammatory agent (e.g., a TLR agonist, e.g., R848, e.g., radiation therapy), and wherein the method comprises administering the SHP-1 inhibitor intravenously or subcutaneously, optionally wherein the SHP-1 inhibitor is administered intermittently. In some embodiments, the SHP-1 inhibitor is administered daily for no more than three or two consecutive days and optionally at least twice (which are separated by at least one day). In some embodiments, the SHP-1 inhibitor is administered at least three, four, or five times. In some embodiments, at least two administrations of the SHP-1 inhibitor are separated by two, three, four, five, six, seven, eight, nine, or ten days. In some embodiments, each administration of the SHP-1 inhibitor is separated by at least one day from the previous or subsequent administration of the SHP-1 inhibitor. In some embodiments, the method comprises administering to the individual at least twice at intervals of no more than once every three days. In some embodiments, the SHP-1 inhibitor is administered twice every seven to twenty days (e.g., two administration days). In some embodiments, the SHP-1 inhibitor is administered three times every ten to twenty days (e.g., three administration days). In some embodiments, the SHP-1 inhibitor is administered no more than once every two days. In some embodiments, the SHP-1 inhibitor is administered no less than twice and no more than five times within ten consecutive 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 SHP-1 inhibitor is administered simultaneously with the proinflammatory agent. In some embodiments, the SHP-1 inhibitor is administered concurrently with the proinflammatory agent. In some embodiments, the SHP-1 inhibitor and the proinflammatory agent are administered sequentially and within two weeks (e.g., within 10 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or within the same day). In some embodiments, the half-life of the SHP-1 inhibitor is 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 SHP-1 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 or activates SHP-1). In some embodiments, the SHP-1 inhibitor is selected from the group consisting of: TPI-1 or an analog or derivative thereof, a vitamin E derivative, Phomopsis xanthone A (PXA), and a PKCθ activator. In some embodiments, the method further comprises administering a pro-inflammatory agent locally (e.g., intratumorally) to the individual.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-TNFα antibody or an anti-IL6 antibody). In some embodiments, the method further comprises administering to the individual an anti-TNFα antibody, optionally wherein the anti-TNFα antibody is administered prior to (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours), concurrently or simultaneously with, or immediately after (within 3, 2, 1, or 0.5 hours) administration of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and / or a proinflammatory agent. In some embodiments, the proinflammatory agent comprises or is selected from the group consisting of: R848, 3M-852A, motomod, bropirimine, and visammod. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0094] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, the method comprising administering to the individual a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a proinflammatory agent (e.g., a TLR agonist, e.g., R848, e.g., radiation therapy), and wherein the method comprises administering the SHP-1 inhibitor intravenously or subcutaneously, optionally wherein the SHP-1 inhibitor is administered intermittently. In some embodiments, the SHP-1 inhibitor is administered daily for no more than three or two consecutive days and optionally at least twice (which are separated by at least one day). In some embodiments, the SHP-1 inhibitor is administered at least three, four, or five times. In some embodiments, at least two administrations of the SHP-1 inhibitor are separated by two, three, four, five, six, seven, eight, nine, or ten days. In some embodiments, each administration of the SHP-1 inhibitor is separated by at least one day from the previous or subsequent administration of the SHP-1 inhibitor. In some embodiments, the method comprises administering a SHP-1 inhibitor to an individual for at least two cycles, further optionally wherein the SHP-1 inhibitor is administered at least once in each cycle and wherein each cycle has a duration of about three to about twenty days. In some embodiments, the SHP-1 inhibitor is administered at least twice in each cycle (e.g., at least two consecutive days). In some embodiments, the SHP-1 inhibitor is administered at least three times in each cycle (e.g., at least three consecutive days). In some embodiments, the SHP-1 inhibitor is administered concurrently with the proinflammatory agent. In some embodiments, the SHP-1 inhibitor is administered concurrently with the proinflammatory agent. In some embodiments, the SHP-1 inhibitor and the proinflammatory agent are administered sequentially and within two weeks (e.g., within 10 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or within the same day). In some embodiments, the half-life of the SHP-1 inhibitor is 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 SHP-1 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 SHP-1 or activate SHP-1). In some embodiments, the SHP-1 inhibitor is selected from the group consisting of: TPI-1 or an analog or derivative thereof, a vitamin E derivative, Phomopsis xanthone A (PXA), and a PKCθ activator. In some embodiments, the method further comprises administering a pro-inflammatory agent locally (e.g., intratumorally) to the individual. 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-TNFα antibody or an anti-IL6 antibody).In some embodiments, the method further comprises administering an anti-TNFα antibody to the individual, optionally wherein the anti-TNFα antibody is administered prior to (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours), concurrently or simultaneously with, or immediately after (within 3, 2, 1, or 0.5 hours) administration of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and / or a proinflammatory agent. In some embodiments, the proinflammatory agent comprises or is selected from the group consisting of: R848, 3M-852A, motomod, bropirimine, and visammod. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0095] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in a subject is provided, the method comprising administering to the subject intravenously, subcutaneously, and / or intratumorally a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a proinflammatory agent (e.g., a TLR agonist, e.g., R848, e.g., radiation therapy), optionally wherein the SHP-1 inhibitor is effective to inhibit SHP-1 activity by greater than 50% for no more than about 5 days, and optionally wherein the method comprises administering the SHP-1 inhibitor to the subject at least twice (e.g., at least 3, 4, 5, or 6 times) at intervals no more than once every three days. In some embodiments, the SHP-1 inhibitor is administered intermittently. In some embodiments, the SHP-1 inhibitor is administered daily for no more than three or two consecutive days and optionally at least twice (which are separated by at least one day). In some embodiments, the SHP-1 inhibitor is administered at least three, four, or five times. In some embodiments, at least two administrations of the SHP-1 inhibitor are separated by two, three, four, five, six, seven, eight, nine, or ten days. In some embodiments, each administration of the SHP-1 inhibitor is separated from the previous or subsequent administration of the SHP-1 inhibitor by at least one day. In some embodiments, the SHP-1 inhibitor is administered no more than twice every seven to twenty days. In some embodiments, the SHP-1 inhibitor is administered no more than three times every seven to twenty days. In some embodiments, the SHP-1 inhibitor is administered about 1-3 times every seven to twenty days for a period of at least fourteen to twenty days. In some embodiments, the SHP-1 inhibitor is administered at least about two, three, four, five, or six times over a period of about fourteen to about forty days (e.g., about fourteen to about twenty days). In some embodiments, the SHP-1 inhibitor is administered simultaneously with the proinflammatory agent. In some embodiments, the SHP-1 inhibitor is administered concurrently with the proinflammatory agent. In some embodiments, the SHP-1 inhibitor and the proinflammatory agent are administered sequentially and within two weeks (e.g., within 10 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or within the same day). In some embodiments, the half-life of the SHP-1 inhibitor is 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 SHP-1 inhibitor effectively inhibits more than 50% of SHP-1 activity for no more than about 7 days (e.g., about 5 days, 4 days, or 3 days). In some embodiments, the SHP-1 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., antibodies that target SHP-1 or activate SHP-1). In some embodiments, the SHP-1 inhibitor is selected from the group consisting of: TPI-1 or an analog or derivative thereof, a vitamin E derivative, Phomopsis xanthonone A (PXA), and a PKCθ activator.In some embodiments, the method further comprises administering a proinflammatory agent locally (e.g., intratumorally) to the individual. In some embodiments, the SHP-1 inhibitor is administered systemically and the proinflammatory agent is administered intratumorally. In some embodiments, the SHP-1 inhibitor is administered systemically and intratumorally. 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-TNFα antibody or an anti-IL6 antibody). In some embodiments, the method further comprises administering to the individual an anti-TNFα antibody, optionally wherein the anti-TNFα antibody is administered prior to (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours), concurrently or simultaneously with, or immediately after (within 3, 2, 1, or 0.5 hours) administration of the SHP-1 inhibitor (e.g., TPI-1 or its analog or derivative) and / or the proinflammatory agent. In some embodiments, the proinflammatory agent comprises or is selected from the group consisting of: R848, 3M-852A, motomod, bropirimine, and visammod. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0096] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, comprising administering to the individual intravenously, subcutaneously, and / or intratumorally a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a proinflammatory agent (e.g., a TLR agonist, e.g., R848, e.g., radiation therapy), wherein the SHP-1 inhibitor is effective to inhibit SHP-1 activity by greater than 50% for no more than about 5 days (e.g., no more than 5, 4, or 3 days), and wherein the SHP-1 inhibitor is administered intermittently. In some embodiments, the SHP-1 inhibitor is administered daily for no more than three or two consecutive days and optionally at least twice (which are separated by at least one day). In some embodiments, the SHP-1 inhibitor is administered at least three, four, or five times. In some embodiments, at least two administrations of the SHP-1 inhibitor are separated by two, three, four, five, six, seven, eight, nine, or ten days. In some embodiments, each administration of the SHP-1 inhibitor is separated by at least one day from the previous or next administration of the SHP-1 inhibitor. In some embodiments, the method comprises administering a SHP-1 inhibitor to an individual for at least two cycles, wherein the SHP-1 inhibitor is administered at least once in each cycle and wherein each cycle has a duration of about three to about twenty days. In some embodiments, the SHP-1 inhibitor is administered at least twice in each cycle (e.g., at least two consecutive days). In some embodiments, the SHP-1 inhibitor is administered at least three times in each cycle (e.g., at least three consecutive days). In some embodiments, the SHP-1 inhibitor is administered simultaneously with the proinflammatory agent. In some embodiments, the SHP-1 inhibitor is administered concurrently with the proinflammatory agent. In some embodiments, the SHP-1 inhibitor and the proinflammatory agent are administered sequentially and within two weeks (e.g., within 10 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or within the same day). In some embodiments, the half-life of the SHP-1 inhibitor is 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 SHP-1 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 or activates SHP-1). In some embodiments, the SHP-1 inhibitor is selected from the group consisting of: TPI-1 or an analog or derivative thereof, a vitamin E derivative, Phomopsis xanthone A (PXA), and a PKCθ activator. In some embodiments, the method further comprises administering a proinflammatory agent locally (e.g., intratumorally) to the individual. In some embodiments, the SHP-1 inhibitor is administered systemically and the proinflammatory agent is administered intratumorally. In some embodiments, the SHP-1 inhibitor is administered systemically and intratumorally.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-TNFα antibody or an anti-IL6 antibody). In some embodiments, the method further comprises administering to the individual an anti-TNFα antibody, optionally wherein the anti-TNFα antibody is administered prior to (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours), concurrently or simultaneously with, or immediately after (within 3, 2, 1, or 0.5 hours) administration of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and / or a proinflammatory agent. In some embodiments, the proinflammatory agent comprises or is selected from the group consisting of: R848, 3M-852A, motomod, bropirimine, and visammod. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0097] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, comprising administering (e.g., intravenously, subcutaneously, and / or intratumorally) a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and an immune cell (e.g., any of the immune cells described herein) to the individual. In some embodiments, the individual has received, is receiving, or will receive a pro-inflammatory 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 a persistent infection. In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, comprising administering (e.g., intravenously, subcutaneously, and / or intratumorally) a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof), a pro-inflammatory agent (e.g., a TLR agonist, e.g., R848, e.g., radiation therapy), and an immune cell to the individual. In some embodiments, the immune cells are derived from the same individual. In some embodiments, the immune cells comprise monocytes or macrophages. In some embodiments, the immune cells comprise T cells (e.g., CAR-T cells). In some embodiments, the immune cells comprise NK cells (e.g., CAR-NK cells). In some embodiments, the immune cells comprise neutrophils (e.g., neutrophil cells expressing CAR). In some embodiments, the immune cells comprise antigen presenting cells (APCs). In some embodiments, the immune cells are engineered to express a chimeric receptor that specifically binds to a tumor antigen. In some embodiments, the SHP-1 inhibitor is administered intermittently. In some embodiments, the SHP-1 inhibitor is administered daily for no more than three or two consecutive days and optionally at least twice (which are at least one day apart). In some embodiments, the SHP-1 inhibitor is administered at least three, four, or five times. In some embodiments, at least two SHP-1 inhibitor administrations are separated by two, three, four, five, six, seven, eight, nine, or ten days. In some embodiments, each administration of the SHP-1 inhibitor is at least one day apart from the previous or subsequent administration of the SHP-1 inhibitor. In some embodiments, the SHP-1 inhibitor, immune cells, and / or proinflammatory agent are administered within 7, 6, 5, 4, 3, 2, or 1 day of each other. In some embodiments, the SHP-1 inhibitor and immune cells are administered within 24 hours of each other (e.g., within 12, 8, 4, 2, or 1 hour, or within 30 minutes). In some embodiments, the SHP-1 inhibitor, immune cells, and / or proinflammatory agent are administered simultaneously. In some embodiments, the SHP-1 inhibitor, immune cells, and / or proinflammatory agent are administered concurrently. In some embodiments, the SHP-1 inhibitor, immune cells, and / or proinflammatory agent are administered sequentially. In some embodiments, the SHP-1 inhibitor is administered systemically and the proinflammatory agent is administered intratumorally.In some embodiments, the SHP-1 inhibitor is administered systemically and intratumorally. 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-TNFα antibody or an anti-IL6 antibody). In some embodiments, the method further comprises administering to the individual an anti-TNFα antibody, optionally wherein the anti-TNFα antibody is administered prior to (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours), concurrently or simultaneously with, or immediately after (within 3, 2, 1, or 0.5 hours) administration of the SHP-1 inhibitor (e.g., TPI-1 or its analog or derivative) and / or proinflammatory agent. In some embodiments, the proinflammatory agent comprises or is selected from the group consisting of: R848, 3M-852A, motomod, bropirimine, and visammod. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0098] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, comprising administering to the individual a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a TLR agonist (e.g., R848), wherein the SHP-1 inhibitor is administered at least twice (e.g., at least 3, 4, or 5 times). In some embodiments, the SHP-1 inhibitor is administered intermittently. In some embodiments, the SHP-1 inhibitor is administered daily for no more than three or two consecutive days and optionally at least twice (which are separated by at least one day). In some embodiments, the SHP-1 inhibitor is administered at least three, four, or five times. In some embodiments, at least two administrations of the SHP-1 inhibitor are separated by two, three, four, five, six, seven, eight, nine, or ten days. In some embodiments, each administration of the SHP-1 inhibitor is separated by at least one day from the previous or subsequent administration of the SHP-1 inhibitor. In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, comprising administering to the individual a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a TLR agonist, wherein the SHP-1 inhibitor and the TLR agonist are administered within 24 hours of each other (e.g., within 12, 8, 4, 2, or 1 hours, or within 30 minutes). In some embodiments, the method comprises administering the SHP-1 inhibitor to the individual at least twice, at intervals no greater than once every three days. In some embodiments, the method comprises administering the SHP-1 inhibitor to the individual for at least two cycles, wherein the SHP-1 inhibitor is administered at least once (e.g., at least two or three times) in each cycle, and wherein each cycle has about three to about twenty days. In some embodiments, the SHP-1 inhibitor is administered systemically (e.g., intravenously or subcutaneously) and / or locally (e.g., intratumorally). In some embodiments, the SHP-1 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, polyI:C, PolyICIC, PolyIC12U, IPH302, ARNAX, and / or MPLA.In some embodiments, the TLR agonist activates TLR4, optionally wherein the TLR agonist comprises LPS, lipoteichoic acid β-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 bistriazole 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 profilin. 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, polyI:C, and / or R848. In some embodiments, the proinflammatory agent comprises the following agents or is selected from the group consisting of: R848, 3M-852A, motomod, bropirimine, and visammod. In some embodiments, the SHP-1 inhibitor is administered systemically and the TLR agonist is administered intratumorally. In some embodiments, the SHP-1 inhibitor is administered systemically and intratumorally. 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-TNFα antibody or an anti-IL6 antibody).In some embodiments, the method further comprises administering to the individual an anti-TNFα antibody, optionally wherein the anti-TNFα antibody is administered prior to (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours), concurrently or simultaneously with, or immediately after (within 3, 2, 1, or 0.5 hours) administration of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and / or a proinflammatory agent. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0099] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, the method comprising administering TPI-1, or an analog or derivative thereof, 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, TPI-1, or an analog or derivative thereof, and the TLR agonist are administered on the same day. In some embodiments, TPI-1, or an analog or derivative thereof, is administered intermittently. In some embodiments, the SHP-1 inhibitor is administered daily for no more than three or two consecutive days and optionally at least twice (separated by at least one day). In some embodiments, the SHP-1 inhibitor is administered at least three, four, or five times. In some embodiments, at least two administrations of the SHP-1 inhibitor are separated by two, three, four, five, six, seven, eight, nine, or ten days. In some embodiments, each administration of the SHP-1 inhibitor is separated by at least one day from the previous or next administration of the SHP-1 inhibitor. In some embodiments, TPI-1 or its analog or derivative and / or TLR agonist are administered at least twice (e.g., at least three, four, five, or six times). In some embodiments, TPI-1 or its analog or derivative and TLR agonist are administered for at least two cycles (e.g., at least three cycles), optionally wherein TPI-1 or its analog or derivative and TLR agonist are administered on 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 to about twenty days. In some embodiments, the TLR agonist activates TLRs on macrophages, optionally wherein the TLRs comprise 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, polyI:C, and / or R848. In some embodiments, the proinflammatory agent includes the following agents or is selected from the group consisting of: R848, 3M-852A, motomod, bropirimine and visammod. In some embodiments, TPI-1 or its analogs or derivatives are administered systemically and the TLR agonist is administered intratumorally. In some embodiments, TPI-1 or its analogs or derivatives are administered systemically and intratumorally. 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-TNFα antibody or an anti-IL6 antibody).In some embodiments, the method further comprises administering to the individual an anti-TNFα antibody, optionally wherein the anti-TNFα antibody is administered prior to (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours), concurrently or simultaneously with, or immediately after (within 3, 2, 1, or 0.5 hours) administration of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and / or a proinflammatory agent. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0100] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, comprising administering to the individual a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a STING activator (e.g., cGAMP, e.g., MSA-2), optionally wherein the SHP-1 inhibitor is administered at least twice (at least three, four, five, or six times). In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, comprising administering to the individual a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a STING activator (e.g., cGAMP, e.g., MSA-2), optionally wherein the SHP-1 inhibitor and the STING activator are administered within 24 hours of each other (e.g., within 12, 8, 4, 2, or 1 hour, or within 30 minutes). In some embodiments, the SHP-1 inhibitor is administered intermittently. In some embodiments, the SHP-1 inhibitor is administered no more than three or two consecutive days and optionally at least twice daily (which are at least one day apart). In some embodiments, the SHP-1 inhibitor is administered at least three, four, or five times. In some embodiments, at least two administrations of the SHP-1 inhibitor are separated by two, three, four, five, six, seven, eight, nine, or ten days. In some embodiments, each administration of the SHP-1 inhibitor is separated by at least one day from the previous or subsequent administration of the SHP-1 inhibitor. In some embodiments, the method comprises administering the SHP-1 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 SHP-1 inhibitor to the individual for at least two cycles, wherein the SHP-1 inhibitor is administered at least once in each cycle and wherein each cycle has a duration of about three to about twenty days. In some embodiments, the SHP-1 inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or locally (e.g., intratumorally). In some embodiments, the SHP-1 inhibitor and the STING activator are administered sequentially, simultaneously, or concurrently.In some embodiments, the STING activator is cyclic-guanosine monophosphate-adenosine monophosphate (cGAMP, such as 3'3'cGAMP, such as 2'3'cGAMP), a bacterial vector (such as SYNB1891, STACT-TREX-1), a CDN compound (such as ADU-S100, BI-STING, BMS-986301, GSK532, JNJ-4412, MK-1454, SB11285, 3'3'-cyclic AIMP), a non-CDN small molecule (such as ALG-031048, E7755, JNJ-'6196, MK-2118, MSA-1, MSA-2, SNX281, SR-717, TAK676, TTI-10001), a nanovaccine (such as PC7A NP, cCAMP-NP, ONM-500) or an antibody-drug conjugate (e.g., XMT-2056, CRD-5500). In some embodiments, the SHP-1 inhibitor is administered systemically and the STING activator is administered intratumorally. In some embodiments, the SHP-1 inhibitor is administered systemically and intratumorally. 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-TNFα antibody or an anti-IL6 antibody). In some embodiments, the method further comprises administering to the individual an anti-TNFα antibody, optionally wherein the anti-TNFα antibody is administered prior to (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours), concurrently or simultaneously with, or immediately after (within 3, 2, 1, or 0.5 hours) administration of the SHP-1 inhibitor (e.g., TPI-1 or its analog or derivative) and / or the proinflammatory agent. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0101] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in a subject is provided, the method comprising administering to the subject a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and radiation therapy, optionally wherein the method comprises administering to the subject the SHP-1 inhibitor for at least two cycles, wherein the SHP-1 inhibitor is administered at least once in each cycle and wherein each cycle has a duration of about three to about twenty days. In some embodiments, the method comprises administering to the subject the SHP-1 inhibitor at least twice, at intervals of no more than once every three days. In some embodiments, the SHP-1 inhibitor is administered at least three times. In some embodiments, the SHP-1 inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or locally (e.g., intratumorally). In some embodiments, the SHP-1 inhibitor and the radiation therapy are administered within 24 hours of each other (e.g., within 12, 8, 4, 2, or 1 hour, or within 30 minutes). In some embodiments, the radiation therapy comprises irradiating the site of the cancer being treated. In some embodiments, the radiation therapy comprises irradiating a site different from the site of the cancer being treated. In some embodiments, the dose of radiotherapy is insufficient to kill tumor cells. In some embodiments, the radiotherapy is selected from the group consisting of external beam radiotherapy, internal radiotherapy (brachytherapy), intraoperative radiotherapy (IORT), systemic radiotherapy, radioimmunotherapy, and the administration of radiosensitizers and radioprotectants. In some embodiments, the radiotherapy is external beam radiotherapy, which optionally includes three-dimensional conformal radiotherapy (3D-RT), intensity modulated radiation therapy (IMRT), photon beam therapy, image-guided radiation therapy (IGRT), and stereotactic radiotherapy (SRT). In some embodiments, the radiotherapy is brachytherapy, which optionally includes interstitial brachytherapy, intracavitary brachytherapy, intraluminal radiotherapy, and intravenously administered radiolabeled molecules. In some embodiments, the SHP-1 inhibitor is administered systemically and intratumorally. 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-TNFα antibody or an anti-IL6 antibody). In some embodiments, the method further comprises administering to the individual an anti-TNFα antibody, optionally wherein the anti-TNFα antibody is administered prior to (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours), concurrently or simultaneously with, or immediately after (within 3, 2, 1, or 0.5 hours) administration of the SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and / or the proinflammatory agent.In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0102] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, the method comprising administering to the individual a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and radiation therapy, wherein the radiation therapy comprises irradiating a site different from the site of the cancer to be treated. In some embodiments, the SHP-1 inhibitor is administered at least twice (at least three, four, five, or six times). In some embodiments, the SHP-1 inhibitor is administered intermittently. In some embodiments, the SHP-1 inhibitor is administered daily for no more than three or two consecutive days and optionally at least twice (separated by at least one day). In some embodiments, the SHP-1 inhibitor is administered at least three, four, or five times. In some embodiments, at least two administrations of the SHP-1 inhibitor are separated by two, three, four, five, six, seven, eight, nine, or ten days. In some embodiments, each administration of the SHP-1 inhibitor is separated by at least one day from the previous or subsequent administration of the SHP-1 inhibitor. In some embodiments, the method comprises administering to the individual at least twice at intervals of no more than once every three days. In some embodiments, the method comprises administering a SHP-1 inhibitor to the individual for at least two cycles, wherein the SHP-1 inhibitor is administered at least once in each cycle and wherein each cycle has a duration of about three to about twenty days. In some embodiments, the SHP-1 inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or locally (e.g., intratumorally). In some embodiments, the SHP-1 inhibitor and the radiation therapy are administered within 24 hours of each other (e.g., within 12, 8, 4, 2, or 1 hours, or within 30 minutes). In some embodiments, the radiation therapy comprises irradiating the site of the cancer to be treated. In some embodiments, the radiation therapy comprises irradiating 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), systemic radiation therapy, radioimmunotherapy, and administration of a radiosensitizer and a radioprotectant. In some embodiments, the radiation therapy is external beam radiation therapy, which optionally includes 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, which optionally includes interstitial brachytherapy, intracavitary brachytherapy, intraluminal radiation therapy, and intravenously administered radiolabeled molecules. In some embodiments, the SHP-1 inhibitor is administered systemically and intratumorally.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-TNFα antibody or an anti-IL6 antibody). In some embodiments, the method further comprises administering to the individual an anti-TNFα antibody, optionally wherein the anti-TNFα antibody is administered prior to (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours), concurrently or simultaneously with, or immediately after (within 3, 2, 1, or 0.5 hours) administration of the SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and / or the proinflammatory agent. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0103] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, the method comprising administering TPI-1, or an analog or derivative thereof, and radiation therapy. In some embodiments, TPI-1, or an analog or derivative thereof, is administered intermittently. In some embodiments, the SHP-1 inhibitor is administered daily for no more than three or two consecutive days and optionally at least twice (separated by at least one day). In some embodiments, the SHP-1 inhibitor is administered at least three, four, or five times. In some embodiments, at least two administrations of the SHP-1 inhibitor are separated by two, three, four, five, six, seven, eight, nine, or ten days. In some embodiments, each administration of the SHP-1 inhibitor is separated by at least one day from the previous or next administration of the SHP-1 inhibitor. In some embodiments, TPI-1, or an analog or derivative thereof, and radiation therapy are administered on the same day. In some embodiments, TPI-1, or an analog or derivative thereof and / or radiation therapy are administered at least twice (e.g., at least three, four, five, or six times). In some embodiments, TPI-1 or its analog or derivative and radiation therapy are administered for at least two cycles (e.g., at least three cycles), optionally wherein TPI-1 or its analog or derivative and radiation therapy are administered on the same day for at least two consecutive days (e.g., at least three consecutive days) in each cycle. In some embodiments, each cycle is about seven to about twenty days. In some embodiments, the SHP-1 inhibitor and radiation therapy are administered within 24 hours of each other (e.g., within 12, 8, 4, 2, or 1 hour, or within 30 minutes). In some embodiments, radiation therapy comprises irradiating the site of the cancer to be treated. In some embodiments, radiation therapy comprises irradiating a site different from the site of the cancer to be treated. In some embodiments, the dose of radiation therapy is insufficient to kill tumor cells. In some embodiments, TPI-1 or its analog or derivative is administered systemically and intratumorally. 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-TNFα antibody or an anti-IL6 antibody). In some embodiments, the method further comprises administering to the individual an anti-TNFα antibody, optionally wherein the anti-TNFα antibody is administered prior to (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours), concurrently or simultaneously with, or immediately after (within 3, 2, 1, or 0.5 hours) administration of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and / or a proinflammatory agent. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0104] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in a subject is provided, comprising administering to the subject a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a PAMP / DAMP activator, optionally wherein the SHP-1 inhibitor is administered at least twice (at least three, four, five, or six times). In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in a subject is provided, comprising administering to the subject a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a PAMP / DAMP activator, optionally wherein the SHP-1 inhibitor and the PAMP / DAMP activator are administered within 24 hours of each other (e.g., within 12, 8, 4, 2, or 1 hour, or within 30 minutes). In some embodiments, the SHP-1 inhibitor is administered intermittently. In some embodiments, the SHP-1 inhibitor is administered no more than three or two consecutive days and optionally at least twice daily (which are separated by at least one day). In some embodiments, the SHP-1 inhibitor is administered at least three, four, or five times. In some embodiments, at least two administrations of the SHP-1 inhibitor are separated by two, three, four, five, six, seven, eight, nine, or ten days. In some embodiments, each administration of the SHP-1 inhibitor is separated by at least one day from the previous or subsequent administration of the SHP-1 inhibitor. In some embodiments, the method comprises administering the SHP-1 inhibitor to the individual at least twice at intervals no more than once every three days. In some embodiments, the method comprises administering the SHP-1 inhibitor to the individual for at least two cycles, wherein the SHP-1 inhibitor is administered at least once in each cycle and wherein each cycle has a duration of about three to about twenty days. In some embodiments, the SHP-1 inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or topically (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 (I: C), trypanosomal lipid, paclitaxel, Pseudomonas exoenzyme S, RSV F protein, MMTV envelope protein, flagellin, diacyl lipopeptide, single-stranded RNA, imiquimod, single-stranded RNA, resquimod, bacterial / viral DNA, CpG DNA, ureobacteria 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 hyaluronic acid, fibrinogen, fibronectin, fx1-defensin, heparan sulfate, HSP60, HSP70, HSP90, HMGB1, or unmethylated CpG DNA. In some embodiments, the SHP-1 inhibitor is administered systemically and the PAMP / DAMP activator is administered intratumorally. In some embodiments, the SHP-1 inhibitor is administered systemically and intratumorally. 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-TNFα antibody or an anti-IL6 antibody). In some embodiments, the method further comprises administering to the individual an anti-TNFα antibody, optionally wherein the anti-TNFα antibody is administered prior to (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours), concurrently or simultaneously with, or immediately after (within 3, 2, 1, or 0.5 hours) administration of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and / or a proinflammatory agent. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0105] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, comprising administering to the individual a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) 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 SHP-1 inhibitor is administered at least twice (at least three, four, five, or six times). In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, comprising administering to the individual a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a checkpoint inhibitor (e.g., an anti-PD-1 agent, an anti-PD-L1 agent, or an anti-CTLA-4 agent), wherein the SHP-1 inhibitor and the checkpoint inhibitor are administered within 24 hours of each other (e.g., within 12, 8, 4, 2, or 1 hour, or within 30 minutes). In some embodiments, the SHP-1 inhibitor is administered intermittently. In some embodiments, the SHP-1 inhibitor is administered daily for no more than three or two consecutive days and optionally at least twice (which are separated by at least one day). In some embodiments, the SHP-1 inhibitor is administered at least three, four, or five times. In some embodiments, at least two administrations of the SHP-1 inhibitor are separated by two, three, four, five, six, seven, eight, nine, or ten days. In some embodiments, each administration of the SHP-1 inhibitor is separated by at least one day from the previous or next administration of the SHP-1 inhibitor. In some embodiments, the method comprises administering the SHP-1 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 SHP-1 inhibitor to the individual for at least two cycles, wherein the SHP-1 inhibitor is administered at least once in each cycle and wherein each cycle has a duration of about three to about twenty days. In some embodiments, the SHP-1 inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or topically (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 / SIRPα, 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 lipilimumab, Cemiplimab, Nivolumab, Pembrolizumab, Atezolizumab, Avelumab, Durvalumab, LAG525 (IMP701), REGN3767, BI 754,091, tebotelimab (MGD013), eftilagimod 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, TT I-662, RRx-001, Lanotuzumab (MCS110), LY3022855, SNDX-6352, Emactuzumab (RG7155), Pexidartinib (PLX3397), CAN04, Canakinumab (ACZ885), BMS-986253, Pepinemab (VX15 / 2503), Trebananib, FP-1305, Enapata monoclonal antibody vedotin (EnaV) or Bavituximab. In some embodiments, the SHP-1 inhibitor is administered systemically and the checkpoint inhibitor is administered intratumorally. In some embodiments, the SHP-1 inhibitor is administered systemically and intratumorally. 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-TNFα antibody or an anti-IL6 antibody). In some embodiments, the method further comprises administering to the individual an anti-TNFα antibody, optionally wherein the anti-TNFα antibody is administered prior to (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours), concurrently or simultaneously with, or immediately after (within 3, 2, 1, or 0.5 hours) administration of the SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and / or the proinflammatory agent. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0106] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, comprising administering to the individual a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a proinflammatory cytokine (e.g., IL-1b, IL-18, IL-6, and / or TNFα), optionally wherein the SHP-1 inhibitor is administered at least twice (at least three, four, five, or six times). In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, comprising administering to the individual a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a proinflammatory cytokine (e.g., IL-1b, IL-18, IL-6, and / or TNFα), wherein the SHP-1 inhibitor and the proinflammatory cytokine are administered within 24 hours of each other (e.g., within 12, 8, 4, 2, or 1 hour, or within 30 minutes). In some embodiments, the SHP-1 inhibitor is administered intermittently. In some embodiments, the SHP-1 inhibitor is administered daily for no more than three or two consecutive days and optionally at least twice (which are separated by at least one day). In some embodiments, the SHP-1 inhibitor is administered at least three, four, or five times. In some embodiments, at least two administrations of the SHP-1 inhibitor are separated by two, three, four, five, six, seven, eight, nine, or ten days. In some embodiments, each administration of the SHP-1 inhibitor is separated by at least one day from the previous or next administration of the SHP-1 inhibitor. In some embodiments, the method comprises administering the SHP-1 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 SHP-1 inhibitor to the individual for at least two cycles, wherein the SHP-1 inhibitor is administered at least once in each cycle and wherein each cycle has a duration of about three to about twenty days. In some embodiments, the SHP-1 inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or topically (e.g., intratumorally). In some embodiments, proinflammatory cytokines promote M1 macrophages. In some embodiments, the proinflammatory cytokine comprises or is TNF, IFNγ and / or GM-CSF. In some embodiments, the proinflammatory cytokine comprises IFNγ. In some embodiments, the proinflammatory cytokine comprises IL-1. In some embodiments, the proinflammatory cytokine comprises TNF-α. In some embodiments, the proinflammatory cytokine comprises IL-6. In some embodiments, the SHP-1 inhibitor is administered systemically and the proinflammatory cytokine is administered intratumorally. In some embodiments, the SHP-1 inhibitor is administered systemically and intratumorally. 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-TNFα antibody or an anti-IL6 antibody).In some embodiments, the method further comprises administering to the individual an anti-TNFα antibody, optionally wherein the anti-TNFα antibody is administered prior to (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours), concurrently or simultaneously with, or immediately after (within 3, 2, 1, or 0.5 hours) administration of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and / or a proinflammatory agent. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0107] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in a subject is provided, comprising administering to the subject a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a chemotherapeutic agent (e.g., azathioprine), optionally wherein the SHP-1 inhibitor is administered at least twice (at least three, four, five, or six times). In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in a subject is provided, comprising administering to the subject a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a chemotherapeutic agent (e.g., azathioprine), wherein the SHP-1 inhibitor and the chemotherapy are administered within 24 hours of each other (e.g., within 12, 8, 4, 2, or 1 hour, or within 30 minutes). In some embodiments, the SHP-1 inhibitor is administered intermittently. In some embodiments, the SHP-1 inhibitor is administered no more than three or two consecutive days and optionally at least twice daily (which are separated by at least one day). In some embodiments, the SHP-1 inhibitor is administered at least three, four, or five times. In some embodiments, at least two administrations of the SHP-1 inhibitor are separated by two, three, four, five, six, seven, eight, nine, or ten days. In some embodiments, each administration of the SHP-1 inhibitor is separated by at least one day from the previous or subsequent administration of the SHP-1 inhibitor. In some embodiments, the method comprises administering the SHP-1 inhibitor to the individual at least twice at intervals no more than once every three days. In some embodiments, the method comprises administering the SHP-1 inhibitor to the individual for at least two cycles, wherein the SHP-1 inhibitor is administered at least once in each cycle and wherein each cycle has a duration of about three to about twenty days. In some embodiments, the SHP-1 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., endamustine, cyclophosphamide, ifosfamide), nitrosoureas (e.g., carmustine, lomustine), platinum analogs (e.g., carboplatin, cisplatin, oxaliplatin), triazenes (e.g., dacarbazine, procarbazine, temozolamide), alkyl sulfonates (e.g., busulfan), and ethyleneimines (e.g., thiotepa). In some embodiments, the chemotherapeutic agent is an antimetabolite.In some embodiments, the antimetabolite is selected from the group consisting of a cytidine analog (e.g., azacitidine, decitabine, cytarabine, gemcitabine), a folate antagonist (e.g., methotrexate, pemetrexed), a purine analog (e.g., cladribine, clofarabine, nelarabine), a pyrimidine analog (e.g., fluorouracil (5-FU), capecitabine (5-FU prodrug)). 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: a topoisomerase II inhibitor (e.g., anthracyclines, doxorubicin, daunorubicin, idarubicin, mitoxantrone), a topoisomerase I inhibitor (e.g., irinotecan, topotecan), a taxane (e.g., paclitaxel, docetaxel, cabazitaxel), a vinca alkaloid (e.g., vinblastine, vincristine, vinorelbine), an antibiotic (e.g., actinomycin, bleomycin, daunomycin). In some embodiments, the chemotherapeutic agent is hydroxyurea, tretinoin, arsenic trioxide, or a proteasome inhibitor (e.g., bortezomib). In some embodiments, the SHP-1 inhibitor is administered systemically and the chemotherapeutic agent is administered intratumorally. In some embodiments, the SHP-1 inhibitor is administered systemically and intratumorally. 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-TNFα antibody or an anti-IL6 antibody). In some embodiments, the method further comprises administering to the individual an anti-TNFα antibody, optionally wherein the anti-TNFα antibody is administered prior to (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours), concurrently or simultaneously with, or immediately after (within 3, 2, 1, or 0.5 hours) administration of the SHP-1 inhibitor (e.g., TPI-1 or its analog or derivative) and / or the proinflammatory agent. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0108] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, comprising administering to the individual a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a cancer vaccine, optionally wherein the SHP-1 inhibitor is administered at least twice. In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, comprising administering to the individual a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a cancer vaccine, wherein the SHP-1 inhibitor and the cancer vaccine are administered within 24 hours of each other (e.g., within 12, 8, 4, 2, or 1 hour, or within 30 minutes). In some embodiments, the SHP-1 inhibitor is administered intermittently. In some embodiments, the SHP-1 inhibitor is administered no more than three or two consecutive days and optionally at least twice daily (which are separated by at least one day). In some embodiments, the SHP-1 inhibitor is administered at least three, four, or five times. In some embodiments, at least two administrations of the SHP-1 inhibitor are separated by two, three, four, five, six, seven, eight, nine, or ten days. In some embodiments, each administration of the SHP-1 inhibitor is separated by at least one day from the previous or subsequent administration of the SHP-1 inhibitor. In some embodiments, the method comprises administering the SHP-1 inhibitor to the individual at least twice, at intervals no greater than once every three days. In some embodiments, the method comprises administering the SHP-1 inhibitor to the individual for at least two cycles, wherein the SHP-1 inhibitor is administered at least once in each cycle and wherein each cycle lasts from about three to about twenty days. In some embodiments, the SHP-1 inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or topically (e.g., intratumorally). In some embodiments, the cancer vaccine comprises a cell-based vaccine, a peptide-based vaccine, a viral-based vaccine, and / or a nucleic acid-based vaccine. In some embodiments, the SHP-1 inhibitor is administered systemically and the cancer vaccine is administered intratumorally. In some embodiments, the SHP-1 inhibitor is administered systemically and intratumorally. 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-TNFα antibody or an anti-IL6 antibody). In some embodiments, the method further comprises administering to the individual an anti-TNFα antibody, optionally wherein the anti-TNFα antibody is administered prior to (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours), concurrently or simultaneously with, or immediately after (within 3, 2, 1, or 0.5 hours) administration of the SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and / or the proinflammatory agent. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0109] In some embodiments, a method for treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, comprising administering a SHP-1 inhibitor (e.g., TPI-1 or its analogs or derivatives) and an oncolytic virus to the individual, optionally wherein the SHP-1 inhibitor is administered at least twice (at least three, four, five, or six times). In some embodiments, a method for treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, comprising administering a SHP-1 inhibitor (e.g., TPI-1 or its analogs or derivatives) and an oncolytic virus to the individual, wherein the SHP-1 inhibitor and the oncolytic virus are administered within 24 hours of each other (e.g., within 12, 8, 4, 2, or 1 hour, or within 30 minutes). In some embodiments, the SHP-1 inhibitor is administered intermittently. In some embodiments, the SHP-1 inhibitor is administered daily for no more than three or two consecutive days and optionally at least twice (which are at least one day apart). In some embodiments, the SHP-1 inhibitor is administered at least three, four, or five times. In some embodiments, at least two administrations of the SHP-1 inhibitor are separated by two, three, four, five, six, seven, eight, nine, or ten days. In some embodiments, each administration of the SHP-1 inhibitor is separated by at least one day from the previous or subsequent administration of the SHP-1 inhibitor. In some embodiments, the method comprises administering the SHP-1 inhibitor to the individual at least twice at intervals no more than once every three days. In some embodiments, the method comprises administering the SHP-1 inhibitor to the individual for at least two cycles, wherein the SHP-1 inhibitor is administered at least once in each cycle and wherein each cycle has a duration of about three to about twenty days. In some embodiments, the SHP-1 inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or topically (e.g., intratumorally). In some embodiments, the oncolytic virus comprises or is an adenovirus (e.g., ONYX-15, LOAd703 virus), a protoparvovirus, 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 (Coxsackievirus A21; CVA21), Maraba virus or its MG1 variant, DNX2440 adenovirus, fowl pox virus, or Sendai virus. In some embodiments, the SHP-1 inhibitor is administered systemically and the oncolytic virus is administered intratumorally. In some embodiments, the SHP-1 inhibitor is administered systemically and intratumorally.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-TNFα antibody or an anti-IL6 antibody). In some embodiments, the method further comprises administering to the individual an anti-TNFα antibody, optionally wherein the anti-TNFα antibody is administered prior to (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours), concurrently or simultaneously with, or immediately after (within 3, 2, 1, or 0.5 hours) administration of the SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and / or the proinflammatory agent. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0110] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, comprising administering to the individual a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and acoustic wave therapy (e.g., high-intensity focused ultrasound (HIFU), e.g., low-intensity focused ultrasound (LIPUS)), optionally wherein the SHP-1 inhibitor is administered at least twice (at least three, four, five, or six times). In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, comprising administering to the individual a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and acoustic wave therapy (e.g., high-intensity focused ultrasound (HIFU), e.g., low-intensity focused ultrasound (LIPUS)), wherein the SHP-1 inhibitor and acoustic wave therapy are administered within 24 hours of each other (e.g., within 12, 8, 4, 2, or 1 hour, or within 30 minutes). In some embodiments, the SHP-1 inhibitor is administered intermittently. In some embodiments, the SHP-1 inhibitor is administered daily for no more than three or two consecutive days and optionally at least twice (which are separated by at least one day). In some embodiments, the SHP-1 inhibitor is administered at least three, four, or five times. In some embodiments, at least two administrations of the SHP-1 inhibitor are separated by two, three, four, five, six, seven, eight, nine, or ten days. In some embodiments, each administration of the SHP-1 inhibitor is separated by at least one day from the previous or next administration of the SHP-1 inhibitor. In some embodiments, the method comprises administering the SHP-1 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 SHP-1 inhibitor to the individual for at least two cycles, wherein the SHP-1 inhibitor is administered at least once in each cycle and wherein each cycle has a duration of about three to about twenty days. In some embodiments, the SHP-1 inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or locally (e.g., intratumorally). In some embodiments, the SHP-1 inhibitor is administered systemically and the method comprises administering acoustic wave therapy to the site of the cancer to be treated. In some embodiments, the SHP-1 inhibitor is administered systemically and intratumorally. 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-TNFα antibody or an anti-IL6 antibody). In some embodiments, the method further comprises administering to the individual an anti-TNFα antibody, optionally wherein the anti-TNFα antibody is administered prior to (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours), concurrently or simultaneously with, or immediately after (within 3, 2, 1, or 0.5 hours) administration of the SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and / or the proinflammatory agent.In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0111] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, comprising administering to the individual a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and magnetic therapy (e.g., a pulsed magnetic field, e.g., a static magnetic field), optionally wherein the SHP-1 inhibitor is administered at least twice (at least three, four, five, or six times). In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, comprising administering to the individual a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and magnetic therapy (e.g., a pulsed magnetic field, e.g., a static magnetic field), wherein the SHP-1 inhibitor and magnetic therapy are administered within 24 hours of each other (e.g., within 12, 8, 4, 2, or 1 hour, or within 30 minutes). In some embodiments, the SHP-1 inhibitor is administered intermittently. In some embodiments, the SHP-1 inhibitor is administered no more than three or two consecutive days and optionally at least twice per day (which are separated by at least one day). In some embodiments, the SHP-1 inhibitor is administered at least three, four, or five times. In some embodiments, at least two administrations of the SHP-1 inhibitor are separated by two, three, four, five, six, seven, eight, nine, or ten days. In some embodiments, each administration of the SHP-1 inhibitor is separated by at least one day from the previous or subsequent administration of the SHP-1 inhibitor. In some embodiments, the method comprises administering the SHP-1 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 SHP-1 inhibitor to the individual for at least two cycles, wherein the SHP-1 inhibitor is administered at least once in each cycle and wherein each cycle has a duration of about three to about twenty days. In some embodiments, the SHP-1 inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or locally (e.g., intratumorally). In some embodiments, the SHP-1 inhibitor is administered systemically and the method comprises administering magnetic therapy to the site of the cancer to be treated. In some embodiments, the SHP-1 inhibitor is administered systemically and intratumorally. 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-TNFα antibody or an anti-IL6 antibody). In some embodiments, the method further comprises administering to the individual an anti-TNFα antibody, optionally wherein the anti-TNFα antibody is administered prior to (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours), concurrently or simultaneously with, or immediately after (within 3, 2, 1, or 0.5 hours) administration of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and / or a proinflammatory agent. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0112] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, comprising administering to the individual a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and electrotherapy or electrochemotherapy, optionally wherein the SHP-1 inhibitor is administered at least twice (at least three, four, five, or six times). In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, comprising administering to the individual a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and electrotherapy or electrochemotherapy, wherein the SHP-1 inhibitor and electrotherapy or electrochemotherapy are administered within 24 hours of each other (e.g., within 12, 8, 4, 2, or 1 hour, or within 30 minutes). In some embodiments, the SHP-1 inhibitor is administered intermittently. In some embodiments, the SHP-1 inhibitor is administered no more than three or two consecutive days and optionally at least twice daily (which are separated by at least one day). In some embodiments, the SHP-1 inhibitor is administered at least three, four, or five times. In some embodiments, at least two administrations of the SHP-1 inhibitor are separated by two, three, four, five, six, seven, eight, nine, or ten days. In some embodiments, each administration of the SHP-1 inhibitor is separated by at least one day from the previous or subsequent administration of the SHP-1 inhibitor. In some embodiments, the method comprises administering the SHP-1 inhibitor to the individual at least twice, at intervals no greater than once every three days. In some embodiments, the method comprises administering the SHP-1 inhibitor to the individual for at least two cycles, wherein the SHP-1 inhibitor is administered at least once in each cycle and wherein each cycle has a duration of about three to about twenty days. In some embodiments, the SHP-1 inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or locally (e.g., intratumorally). In some embodiments, the SHP-1 inhibitor is administered systemically and the method comprises administering electrotherapy or electrochemotherapy to the site of the cancer to be treated. In some embodiments, the SHP-1 inhibitor is administered systemically and intratumorally. 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-TNFα antibody or an anti-IL6 antibody). In some embodiments, the method further comprises administering to the individual an anti-TNFα antibody, optionally wherein the anti-TNFα antibody is administered prior to (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours), concurrently or simultaneously with, or immediately after (within 3, 2, 1, or 0.5 hours) administration of the SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and / or the proinflammatory agent. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0113] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in a subject is provided, comprising administering to the subject a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and electrostatic therapy, optionally wherein the SHP-1 inhibitor is administered at least twice (at least three, four, five, or six times). In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in a subject is provided, comprising administering to the subject a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and electrostatic therapy, wherein the SHP-1 inhibitor and electrostatic therapy are administered within 24 hours of each other (e.g., within 12, 8, 4, 2, or 1 hour, or within 30 minutes). In some embodiments, the SHP-1 inhibitor is administered intermittently. In some embodiments, the SHP-1 inhibitor is administered no more than three or two consecutive days and optionally at least twice daily (which are separated by at least one day). In some embodiments, the SHP-1 inhibitor is administered at least three, four, or five times. In some embodiments, at least two administrations of the SHP-1 inhibitor are separated by two, three, four, five, six, seven, eight, nine, or ten days. In some embodiments, each administration of the SHP-1 inhibitor is separated by at least one day from the previous or subsequent administration of the SHP-1 inhibitor. In some embodiments, the method comprises administering the SHP-1 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 SHP-1 inhibitor to the individual for at least two cycles, wherein the SHP-1 inhibitor is administered at least once in each cycle and wherein each cycle has a duration of about three to about twenty days. In some embodiments, the SHP-1 inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or topically (e.g., intratumorally). In some embodiments, the SHP-1 inhibitor is administered systemically and the method comprises administering electrostatic therapy to the site of the cancer to be treated. In some embodiments, the SHP-1 inhibitor is administered systemically and intratumorally. 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-TNFα antibody or an anti-IL6 antibody). In some embodiments, the method further comprises administering to the individual an anti-TNFα antibody, optionally wherein the anti-TNFα antibody is administered prior to (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours), concurrently or simultaneously with, or immediately after (within 3, 2, 1, or 0.5 hours) administration of a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and / or a proinflammatory agent. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0114] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematologic cancer, e.g., an advanced cancer) in a subject is provided, comprising administering to the subject a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof), wherein the subject is selected for treatment based on having a persistent inflammatory response. In some embodiments, the subject has an acute inflammatory response. In some embodiments, the inflammatory response is localized in the tumor. In some embodiments, the inflammatory response is localized at a site distinct from the tumor. In some embodiments, a subject 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 (e.g., IFNγ, IL-12b, TNFα, 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 (e.g., TGFb1, TGFb2, TGFb3), c) an increase in infiltrating immune cells (e.g., T cells, NK cells, macrophages, neutrophils), d) a decrease in suppressive immune cells (e.g., MDSCs), and / or e) an increase in one or more (e.g., at least one, two, three, four, or five) immunogenic co-stimulatory molecules (e.g., CD80, CD86, OX40L, CD40, ICOS-L, PD-L1, GITRL) in tissue (e.g., tumor tissue) or immune cells (e.g., macrophages). In some embodiments, the SHP-1 inhibitor is administered intermittently. In some embodiments, the SHP-1 inhibitor is administered daily for no more than three or two consecutive days and optionally at least twice (separated by at least one day). In some embodiments, the SHP-1 inhibitor is administered at least three, four, or five times. In some embodiments, at least two administrations of the SHP-1 inhibitor are separated by two, three, four, five, six, seven, eight, nine, or ten days. In some embodiments, each administration of the SHP-1 inhibitor is separated by at least one day from the previous or subsequent administration of the SHP-1 inhibitor. In some embodiments, the SHP-1 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 or activates SHP-1). In some embodiments, the SHP-1 inhibitor is selected from the group consisting of TPI-1 or an analog or derivative thereof, a vitamin E derivative, Phomopsis xanthonone A (PXA), and a PKCθ activator. In some embodiments, the SHP-1 inhibitor is administered at least twice (eg, at least three, four, five, or six times). In some embodiments, the method comprises administering the SHP-1 inhibitor to the individual at least twice at intervals no more than once every three days.In some embodiments, the method comprises administering a SHP-1 inhibitor to the individual for at least two cycles, wherein the SHP-1 inhibitor is administered at least once in each cycle and wherein each cycle is about three to about twenty days. In some embodiments, the SHP-1 inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or locally (e.g., intratumorally). 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-TNFα antibody or an anti-IL6 antibody). In some embodiments, the method further comprises administering to the individual an anti-TNFα antibody, optionally wherein the anti-TNFα antibody is administered prior to (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours), concurrently or simultaneously with, or immediately after (within 3, 2, 1, or 0.5 hours) administration of the SHP-1 inhibitor (e.g., TPI-1 or its analog or derivative) and / or the proinflammatory agent. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0115] In some embodiments, a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual is provided, the method comprising administering a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) to the individual, wherein the individual is selected for treatment based on the individual having persistent 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 DAMPs (e.g., at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%) than a reference sample (e.g., a corresponding sample from a healthy control, e.g., a sample from the cancer before administration of a therapy that induces ICD). In some embodiments, the SHP-1 inhibitor is administered intermittently. In some embodiments, the SHP-1 inhibitor is administered no more than three or two consecutive days and optionally at least twice daily (which are separated by at least one day). In some embodiments, the SHP-1 inhibitor is administered at least three, four, or five times. In some embodiments, at least two administrations of the SHP-1 inhibitor are separated by two, three, four, five, six, seven, eight, nine, or ten days. In some embodiments, each administration of the SHP-1 inhibitor is separated by at least one day from the previous or subsequent administration of the SHP-1 inhibitor. In some embodiments, the DAMP is selected from the group consisting of: endoplasmic reticulum (ER) chaperone proteins (e.g., calreticulin (CALR), such as 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 SHP-1 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., antibodies that target SHP-1 or activate SHP-1). In some embodiments, the SHP-1 inhibitor is selected from the group consisting of: TPI-1 or an analog or derivative thereof, a vitamin E derivative, Phomopsis xanthone A (PXA), and a PKCθ activator. In some embodiments, the SHP-1 inhibitor is administered at least twice (e.g., at least three, four, five, or six times). In some embodiments, the method comprises administering the SHP-1 inhibitor to the individual at least twice at intervals no more than once every three days. In some embodiments, the method comprises administering the SHP-1 inhibitor to the individual for at least two cycles, wherein the SHP-1 inhibitor is administered at least once in each cycle and wherein each cycle has a duration of about three to about twenty days. In some embodiments, the SHP-1 inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and / or topically (e.g., intratumorally).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-TNFα antibody or an anti-IL6 antibody). In some embodiments, the method further comprises administering to the individual an anti-TNFα antibody, optionally wherein the anti-TNFα antibody is administered prior to (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours), concurrently or simultaneously with, or immediately after (within 3, 2, 1, or 0.5 hours) administration of the SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and / or the proinflammatory agent. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0116] In some embodiments, the present application provides a method of treating cancer (e.g., a solid tumor, e.g., a hematological cancer, e.g., an advanced cancer) in an individual, comprising administering to the individual a) monocytes or macrophages defective in SHP-1 expression or activation, and b) a proinflammatory agent (e.g., a TLR agonist, e.g., R848, e.g., radiation therapy). In some embodiments, the monocytes or macrophages are derived from the same individual. In some embodiments, the monocytes or macrophages are engineered to express a chimeric receptor targeting a tumor antigen. In some embodiments, the monocytes or macrophages and the proinflammatory agent are administered within 24 hours of each other (e.g., within 12, 8, 4, 2, or 1 hour, or within 30 minutes). In some embodiments, the monocytes or macrophages and the proinflammatory agent are administered simultaneously, concurrently, or sequentially. In some embodiments, the monocytes or macrophages are administered before the proinflammatory agent. In some embodiments, the monocytes or macrophages are administered after the proinflammatory agent. 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-TNFα antibody or an anti-IL6 antibody). In some embodiments, the method further comprises administering to the individual an anti-TNFα antibody, optionally wherein the anti-TNFα antibody is administered prior to (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours), concurrently or simultaneously with, or immediately after (within 3, 2, 1, or 0.5 hours) administration of the SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and / or the proinflammatory agent. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0117] The present application also provides a method for regulating monocytes or macrophages derived from an individual suffering from cancer, the method comprising contacting the monocytes or macrophages with a SHP-1 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 an agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFα antibody or an anti-IL6 antibody). In some embodiments, the method further comprises administering to the individual an anti-TNFα antibody, optionally wherein the anti-TNFα antibody is administered prior to (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours), concurrently or simultaneously with, or immediately after (within 3, 2, 1, or 0.5 hours) administration of the SHP-1 inhibitor (e.g., TPI-1 or its analog or derivative) and / or the proinflammatory agent. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0118] The present application also provides a method for activating phagocytosis of tumor cells in an individual with a tumor, the method comprising administering a SHP-1 inhibitor to the individual, wherein the individual a) has received, is receiving, or will receive a pro-inflammatory agent, or b) is in an inflammatory response or has a persistent infection. In some embodiments, the SHP-1 inhibitor is administered systemically (e.g., intravenously or subcutaneously). The present application also provides a method for activating tumor-infiltrating T cells in an individual with a tumor, the method comprising administering a SHP-1 inhibitor to the individual, wherein the individual a) has received, is receiving, or will receive a pro-inflammatory agent, or b) is in an inflammatory response or has a persistent infection. In some embodiments, the method comprises administering the SHP-1 inhibitor to the individual at least twice at intervals no more than once every three days. In some embodiments, the method comprises administering the SHP-1 inhibitor to the individual for at least two cycles, wherein the SHP-1 inhibitor is administered at least once in each cycle and wherein each cycle has a duration of about three to about twenty days. In some embodiments, the pro-inflammatory agent and the SHP-1 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, radiotherapy, 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 an agent that reduces systemic inflammation and / or reduces the inflammatory cytokine cascade or cytokine storm (e.g., an anti-TNFα antibody or an anti-IL6 antibody). In some embodiments, the method further comprises administering to the individual an anti-TNFα antibody, optionally wherein the anti-TNFα antibody is administered before (e.g., within two weeks, ten days, one week, 48 hours, or 24 hours) to, concurrently with, or immediately thereafter (within 3, 2, 1, or 0.5 hours) to, a SHP-1 inhibitor (e.g., TPI-1, or its analogs or derivatives) and / or a proinflammatory agent. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0119] It was also found that the strong inhibitory regulation by iR-SHP-1 within the tumor was largely dependent on the physical contact between cancer cells and macrophages. Using transwells that allow the transfer of soluble factors but prevent cancer cells from "contacting" macrophages to separate cancer cells from macrophages failed to exert a strong inhibitory effect on the proinflammatory response of macrophages. See, for example, Figure 19. Combining iR blockers (e.g., antibodies, fusion proteins, or other agents that block the following: a) SIGLEC interaction with sialic acid proteoglycans, b) LILRB or MHC for interaction, c) CD47 or SIRPα, d) lectin receptors, or e) signaling lymphocytic activation molecule family (SLAMF) receptors or their ligands) also achieved the effect of eliminating iR-mediated inhibition, thereby allowing macrophage activation in a proinflammatory direction, through Figure 27C This was demonstrated by the elevation of cytokines shown in .

[0120] Therefore, it is also contemplated that iR blockers (e.g., antibodies, fusion proteins, or other agents that block the interaction between SIGLEC and sialoproteoglycan ligands, block the interaction between LILRB and MHC, block the interaction between CD47 and SIRPα, block the interaction between lectins and lectin receptors, block the interaction between signaling lymphocytic activation molecule family (SLAMF) receptors and their ligands, etc.), and particularly combinations of these blockers, can be used in place of SHP-1 inhibitors in the methods described herein. See, e.g., Figure 27C In some embodiments, a method of treating cancer is provided, the method comprising administering at least two or three blocking agents that block different interactions selected from the group consisting of: SIGLEC-sialoproteoglycan, LILRB-MHC, CD47-SIRPα, lectin-lectin receptor, signaling lymphocytic activation molecule family (SLAMF) receptor-its ligand; and optionally a pro-inflammatory agent.

[0121] Blockers described herein include any agent that: a) reduces the binding of an inhibitory receptor to its ligand, as measured by, for example, spectrometry, isothermal titration calorimetry (ITC), optical biosensors (e.g., surface plasmon resonance (SPR)), biolayer interferometry (BLI), or grating coupled interferometry (GCI), and / or b) activates the inhibitory receptor, as measured by, for example, Western blot activation of downstream signaling by at least 30%, 40%, 50%, 60%, 70%, 80%, or 90%. Exemplary blocking agents include, for example, blocking antibodies that bind to an iR or its ligand.

[0122] In some embodiments, the method comprises administering to an individual in need thereof a) a blocker of CD47-SIRPα (e.g., an anti-CD47 antibody (e.g., B6H12) or an anti-SIRPα antibody), b) a blocker of LILRB-MHC (e.g., an antibody to LILRB1, LILRB2, and / or LILRB3, e.g., an antibody to HLA-A, HLA-B, and / or HLA-C), c) a blocker of SIGLEC-sialoproteoglycans (e.g., anti-siglec9, anti-siglec7, anti-siglec8, e.g., neuraminidase), and optionally d) a pro-inflammatory agent (e.g., a TLR agonist, a STING activator). In some embodiments, the method comprises administering to an individual in need thereof a) a neuraminidase, b) an anti-CD47 antibody, c) an anti-HLA-A / B / C, and optionally d) a pro-inflammatory agent (e.g., a TLR agonist, a STING activator). Exemplary antibodies to iRs or their ligands can be found in Figure 27B In some embodiments, the individual has a persistent infection and does not require treatment with a pro-inflammatory agent.

[0123] Tumor microenvironment (TME) immunosuppression and SHP-1 signaling

[0124] 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. It contains two promoter regions (within exons 1 and 2) that produce two SHP-1 forms with different N-terminal amino acid sequences but similar phosphatase activities. Promoter I is active in non-hematopoietic cells, while promoter II is active in hematopoietic cells. In some epithelial cancer cells, both promoters are functional and produce multiple alternative SHP-1 transcripts. The two SHP-1 isoforms exhibit distinct subcellular localizations: form I is primarily localized in the nucleus, while form II is located in the cytoplasm, suggesting that they have different targets.

[0125] SHP-1 is a 595-amino acid protein composed of two tandem N-terminal SH2 domains (N-SH2 and C-SH2), a typical catalytic protein tyrosine phosphatase (PTP) domain, and a C-terminal tail containing several phosphorylation sites. Its crystals reveal a structure in which N-SH2 binds to the protein's catalytic site via charge-charge interactions. In this autoinhibited, inactive state, substrates are prevented from accessing the active site, but binding of phosphotyrosine residues to the SH2 domains induces conformational changes that weaken the interaction between N-SH2 and the catalytic domain. This opens the conformation to allow substrate access and is further stabilized by new interactions between the SH2 domains and the catalytic domain. These molecular rearrangements determine a complex regulatory mechanism controlled by substrate recruitment.

[0126] Another activation mechanism is mediated by phosphorylation of amino acids within the C-terminal tail. To date, three phosphorylation sites have been identified: two tyrosine (Tyr536 and Tyr564) and one serine (Ser591) residue. Tyr536 and Tyr564 become phosphorylated following various stimuli (i.e., insulin stimulation or apoptosis-inducing agents), leading to increased SHP-1 activity. The molecular mechanism is unclear, but it has been proposed that Tyr phosphorylation may lead to interaction with the N-SH2 domain, thereby releasing this domain's inhibitory effect on PTPase activity. SHP-1 activity can also be negatively regulated by protein kinase C (PKC) or mitogen-activated protein kinase (MAPK) via phosphorylation at Ser591, but the inhibitory mechanism has not been fully characterized.

[0127] Protein-tyrosine phosphorylation is a reversible post-translational modification that is tightly regulated by both kinases and phosphatases. Any deviation in 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, primarily involved in the steady-state maintenance of all these processes. SHP-1 function is actually altered in solid cancers and hematological human cancers via somatic mutations or epigenetic mechanisms. In addition to its well-documented role in regulating hematopoietic cell biology, SHP-1 has now been implicated in a variety of signal transduction pathways related to cancer pathogenesis and progression.

[0128] However, inhibition of SHP-1 causes severe side effects. v / me v ) revealed key immunological abnormalities and overactivation of immune cells associated with global loss of SHP-1 (6,7). Wormed mice often succumb to life-threatening autoimmune inflammatory disorders at an early age. Even partial depletion of SHP-1 in wild-type mice induces features of an inflammatory disease as they grow to adulthood, resulting in widespread lung inflammation and splenomegaly. Like a double-edged sword, despite its potential for anticancer immunity, inhibition of SHP-1 inevitably compromises the host’s inflammatory response, cytokine storms, and enhanced autoimmunity.

[0129] Inhibitors targeting SHP-1 phosphatase activity have been under development for some time, and several have now entered preclinical studies, including NSC-87877, sodium antimony gluconate (SSG), tyrosine phosphatase inhibitor 1 (TPI-1 or its analogs or derivatives), and suramin; however, only a few of these have shown activity in experimental tumor models. SSG has passed Phase I trials for both malignant melanoma (NCT00498979) and advanced malignancies (NCT00629200); the drug was administered in combination with interferon, followed by chemotherapy or without chemotherapy. Unfortunately, no effect on tumor progression was seen, with the most common toxic side effects being thrombocytopenia, elevated serum lipase, fatigue, fever, chills, anemia, hypokalemia, pancreatitis, and rash (observed in up to 68% of patients). Currently, no SHP-1 inhibitor is in Phase II trials.

[0130] Agents that reduce systemic inflammation

[0131] In some cases, individuals develop systemic inflammation, i.e., cytokine release syndrome (CRS), after receiving, for example, immunotherapy treatment, but inflammatory conditions are not fully understood. CRS may be induced by direct target cell lysis and continuous release of cytokines such as TNFα or IFNγ or by T cell activation due to therapeutic stimulation after subsequent cytokine release. Due to the activation of innate immune cells such as macrophages and endothelial cells, these cytokines trigger a chain reaction and then induce further cytokine release. In particular, IL6, IL10, and IFNγ are most commonly found to be elevated in patients with CRS.

[0132] The methods described herein may also include administering an agent that reduces systemic inflammation (including, for example, an agent that reduces the inflammatory cytokine cascade or cytokine storm) to suppress systemic inflammation and reduce adverse toxicity. Agents that reduce systemic inflammation include, but are not limited to, inhibitors of TNFα, IL6, IL10, and IFNγ. In some embodiments, the agent that reduces systemic inflammation is administered simultaneously with the SHP-1 inhibitor. In some embodiments, the agent that reduces systemic inflammation is administered sequentially with (e.g., before or after) the SHP-1 inhibitor. In some embodiments, administration of the agent that reduces systemic inflammation follows the same dosing schedule as the SHP-1 inhibitor. In some embodiments, the agent that reduces systemic inflammation 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, administration of an agent that reduces systemic inflammation allows for more frequent administration of the SHP-1 inhibitor and / or proinflammatory agent (e.g., daily, once every two days, once every three days, etc.).

[0133] The medicament may include any anti-inflammatory agent known in the art, including inhibitors of proinflammatory agents or antagonists of proinflammatory agents. For example, the medicament may 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, derivatives thereof, and any combination thereof, including combinations of medicaments that target one or more molecules (e.g., via inhibition of TNFα alone, IL6 alone, or a combination of TNFα and IL6).

[0134] Anti-TNFα antagonists

[0135] The major proinflammatory cytokine TNFα is secreted by activated macrophages, monocytes, and lymphocytes. The inventors unexpectedly discovered that administering an anti-TNFα antibody to an individual already administered a SHP-1 inhibitor and a proinflammatory agent alleviates toxicity caused by systemic inflammation without compromising the efficacy of the therapeutic agents.

[0136] Thus, in some embodiments, the methods of the present application include administering a TNFα inhibitor, such as an anti-TNFα antagonist (e.g., in a context where the proinflammatory agent is not TNFα). In some embodiments, the TNFα inhibitor is selected from the group consisting of: small molecule inhibitors, neutralizing antibodies, TNFα receptor blocking antibodies, soluble TNFα receptors, short interfering RNA (siRNA) targeting TNFα, chemical inhibitors of TNFα mRNA stability, inhibitors of TNFα converting enzyme (TACE), and derivatives thereof. In some embodiments, the TNFα inhibitor is an anti-TNFα neutralizing antibody. In some embodiments, the TNFα inhibitor is an anti-TNFα receptor blocking antibody. In some embodiments, the anti-TNFα antibody is a monoclonal antibody. In some embodiments, the anti-TNFα antibody is a chimeric, humanized, and / or fully human antibody.

[0137] Antibodies suitable for use in the methods provided herein include, but are not limited to: (Infliximab (Centocor)); and those antibodies described, for example, in U.S. Pat. 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. 0 610 201, the contents of each of which are hereby incorporated by reference in their entirety; or with Antibodies that bind to the same epitope. By way of non-limiting example, other anti-TNFα antibodies suitable for use in the methods provided herein are: Humira (Adalimumab (Abbott Laboratories, Esai)), 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 which are hereby incorporated by reference in their entirety; Simponi TM (Golimimab, CNTO 148 (Centocor)), as described in PCT Publication No. WO 02 / 12502 and related patents and applications, the contents of which are hereby incorporated by reference in their entireties; ART621 (Arana Therapeutics), SSS 07 (Epitopmics and 3SBio), or an antibody that binds to the same epitope as Humira, Simponi, ART621, or SSS 07.

[0138] In some embodiments, the TNFα inhibitor (e.g., an anti-TNFα antagonist) is a fusion protein. Fusion proteins suitable 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.

[0139] In some embodiments, the TNFα inhibitor (e.g., anti-TNFα 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 (Certolizumab pegol, CDP870 (Enzon)), bispecific antibodies, (e.g., ABX 0402 (Ablynx)), immunotoxins and radiolabeled therapeutics; peptide therapeutics; gene therapy, particularly intracellular antibodies; 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.

[0140] In some embodiments, the TNFα inhibitor (e.g., an anti-TNFα antibody) is administered within two weeks, 10 days, or one week prior to administration of a SHP-1 inhibitor and / or proinflammatory agent described herein. Exemplary TNFα inhibitors (e.g., anti-TNFα antibodies) are generally stable for at least one or two weeks. In some embodiments, the TNFα inhibitor (e.g., an anti-TNFα antibody) is administered concurrently or simultaneously with the SHP-1 inhibitor and / or proinflammatory agent. In some embodiments, the TNFα inhibitor (e.g., an anti-TNFα antibody) is administered immediately (e.g., within 1 hour or 30 minutes) after administration of the SHP-1 inhibitor and / or proinflammatory agent.

[0141] In some embodiments, the TNFα inhibitor is administered systemically. In some embodiments, the TNFα inhibitor is administered at least once a week, once every five days, once every three days, or every day. In some embodiments, the TNFα inhibitor is administered intermittently. In some embodiments, the TNFα inhibitor is administered to the individual for at least two cycles, wherein each cycle has about three to about seven days. In some embodiments, the individual does not develop cytokine release syndrome or proinflammatory organ damage. In some embodiments, administration of the TNFα inhibitor does not impair or minimally impairs tumor clearance.

[0142] Anti-IL6 antagonists

[0143] "Anti-IL6 antagonists" or "IL6 inhibitors" refer to agents that inhibit or block the biological activity of IL6 by binding to IL6 or IL6 receptors. In some embodiments, the anti-IL6 antagonist is an antibody. In one embodiment, the anti-IL6 antagonist is an antibody that binds to the IL6 receptor. Antibodies that bind to the IL-6 receptor include tocilizumab (including its intravenous iv and subcutaneous sc 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 to IL6. Antibodies that bind to IL-6 include sirukumab (Centecor, Janssen), olokizumab (UCB), clazakizumab (BMS and Alder), siltuximab (Janssen), and EBI-031 (Eleven Biotherapeutics and Roche). In one embodiment, the IL6 antagonist is olamkicept.

[0144] 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 daily. In some embodiments, the IL6 inhibitor is administered intermittently. In some embodiments, the IL6 inhibitor is administered to an individual for at least two cycles, wherein each cycle has about three to about seven days.

[0145] SHP-1 inhibitors

[0146] As used herein, a SHP-1 inhibitor is any type or class of agent that inhibits the expression or activation of SHP-1. In some embodiments, the SHP-1 inhibitor directly targets SHP-1. In some embodiments, the SHP-1 inhibitor targets a molecule other than SHP-1 that is involved in the SHP-1 signaling pathway in macrophages.

[0147] In some embodiments, the SHP-1 inhibitor is capable of inhibiting SHP-1 activity by at least about 20% (e.g., at least 20%, 30%, 40%, or 50%). In some embodiments, the SHP-1 inhibitor is capable of inhibiting SHP-1 expression by at least about 20% (e.g., at least 20%, 30%, 40%, or 50%).

[0148] In some embodiments, the SHP-1 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), protein agents (e.g., antibody agents that target SHP-1 or activate SHP-1, such as dominant negative SHP-1 or constitutively active SHP-1 mutants), SH2 domain-containing protein agents (which inhibit SHP-1 activation by competing for binding to ITIM motifs), and tyrosine kinase inhibitors that inhibit ITIM phosphorylation.

[0149] In some embodiments, the SHP-1 inhibitor does not significantly inhibit SHP-2 (eg, does not inhibit SHP-2 activity by more than 50%, 40%, 30%, or 20%).

[0150] In some embodiments, the SHP-1 inhibitor also inhibits SHP-2.

[0151] In some embodiments, the half-life of the SHP-1 inhibitor is no more than about 10, 9, 8, or 7 days (eg, a half-life of no more than about 7, 6, 5, 4, 3, 2, or 1 day).

[0152] In some embodiments, the SHP-1 inhibitor is effective to inhibit SHP-1 activity by greater than 50% for no more than about 10, 9, 8, 7, 6, or 5 days. In some embodiments, the SHP-1 inhibitor is effective to inhibit SHP-1 activity by greater than 50% for no more than 4, 3, 2, or 1 day.

[0153] In some embodiments, the SHP-1 inhibitor is a covalent inhibitor. In some embodiments, the SHP-1 inhibitor is a non-covalent inhibitor.

[0154] In some embodiments, the SHP-1 inhibitor is a competitive inhibitor. In some embodiments, the SHP-1 inhibitor is Phomopsis xanthone A (PXA) or Phomopsis xanthone B (PXB). See, e.g., Yang et al., ACSOmega. 2020 Sep 29;5(40):25927-25935

[0155] In some embodiments, the SHP-1 inhibitor targets the catalytic site. In some embodiments, the SHP-1 inhibitor binds to the catalytic site (e.g., covalently or competitively). Exemplary catalytic site inhibitors include TPI-1 or TPI analogs, such as those shown in Kundu et al. (e.g., TPI-1a1-10). See J Immunol. 2010 Jun 1;184(11):6529-6536. Methods for screening and identifying SHP-1 inhibitors (e.g., SHP-1 inhibitors that target the catalytic site) are known in the art. For example, recombinant proteins of the SHP-1 catalytic domain can be used to screen and identify SHP-1 inhibitors that target the catalytic site. SHP-1 inhibitory activity can be evaluated using a variety of methods (e.g., the rapid SHP-1 PTP assay). See "Materials and Methods" in Kundu et al.

[0156] In some embodiments, the SHP-1 inhibitor targets an ectopic or regulatory site. For the structure of SHP-1, see, eg, Wang et al., J Cell Biochem. 2011 Aug; 112(8):2062-2071.

[0157] In some embodiments, the SHP-1 inhibitor is TPI-1, a derivative thereof, or an analog thereof. Exemplary analogs include those disclosed in Kundu et al. (J Immunol. 2010 Jun 1; 184(11): 6529-6536). See, for example, Figure 6 of Kundu et al.

[0158] In some embodiments, the SHP-1 inhibitor comprises TPI-1.

[0159] In some embodiments, the SHP-1 inhibitor is PTP-I.

[0160] In some embodiments, the SHP-1 inhibitor is vitamin E. In some embodiments, the SHP-1 inhibitor is tocofersolan (TPGS). In some embodiments, the SHP-1 inhibitor is alpha-tocopheryl acetate (αTA). In some embodiments, the SHP-1 inhibitor is alpha-tocopheryl succinate (αTOS).

[0161] In some embodiments, the SHP-1 inhibitor is Phomopsis xanthone A (PXA).

[0162] In some embodiments, the SHP-1 inhibitor is a PKC theta activator (eg, PMA).

[0163] In some embodiments, the SHP-1 inhibitor is an siRNA or shRNA that inhibits or knocks down the amount of endogenous SHP-1 protein. See, for example, WO2009 / 023333.

[0164] In some embodiments, the SHP-1 inhibitor is a dominant negative SHP-1 or a constitutively active SHP-1 mutant. See, for example, WO2009 / 023333.

[0165] In some embodiments, the SHP-1 inhibitor is a nucleic acid editing system (e.g., a CRISPR system). In some embodiments, CRISPR components are introduced into cells (e.g., monocytes and macrophages), but the DNA encoding the guide RNA or Cas9 is not incorporated into the cell's genome. In this approach, the CRISPR system only cleaves the cell's genomic DNA for a limited period of time. See, e.g., Fister et al., Front Plant Sci. 2018 Mar 2; 9:268.

[0166] In some embodiments, the SHP-1 inhibitor is a chemical inducer of dimerization. See, e.g., Buck et al., ACS Omega. 2022 Apr 11;7(16):14180-14188.

[0167] In some embodiments, the SHP-1 inhibitor (eg, TPI-1 or an analog or derivative thereof) is administered at least twice (eg, at least 3, 4, 5, or 6 times).

[0168] In some embodiments, the method comprises administering the SHP-1 inhibitor (eg, TPI-1 or an analog or derivative thereof) at least two times (eg, at least three, four, five, or six times) at intervals no more than once every two days.

[0169] In some embodiments, the method comprises administering the SHP-1 inhibitor (eg, TPI-1 or an analog or derivative thereof) at least two times (eg, at least three, four, five, or six times) at intervals no more than once every three days.

[0170] In some embodiments, the methods comprise administering a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) for at least two cycles. In some embodiments, the SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) is administered at least once (e.g., twice, three times, four times) in each cycle. In some embodiments, each cycle has about three to about 50 days (e.g., about 3-40 days, about 3-30 days, about 3-20 days, about 3-15 days, about 3-10 days, or about 2-10 days).

[0171] In some embodiments, the SHP-1 inhibitor is administered systemically (e.g., orally, intravenously, subcutaneously, intraperitoneally). In some embodiments, the SHP-1 inhibitor is administered locally (e.g., intratumorally). In some embodiments, the SHP-1 inhibitor is administered both systemically and locally (e.g., intratumorally).

[0172] In some embodiments, the SHP-1 inhibitor is complexed with a delivery vehicle prior to administration to a subject. In some embodiments, the delivery vehicle facilitates delivery to a tumor.

[0173] In some embodiments, the SHP-1 inhibitor modulates monocytes or macrophages in vitro (eg, monocytes or macrophages derived from the individual to be treated).

[0174] In some embodiments, the SHP-1 inhibitor and the proinflammatory agent are administered within 24 hours of each other (e.g., within 12, 8, 4, 2, or 1 hours, or within 30 minutes). In some embodiments, the SHP-1 inhibitor and the proinflammatory agent are administered simultaneously, concurrently, or sequentially. In some embodiments, the SHP-1 inhibitor is administered before the proinflammatory agent. In some embodiments, the SHP-1 inhibitor is administered after the proinflammatory agent.

[0175] Proinflammatory agents

[0176] Infection and tissue damage are two typical inflammatory factors (instigator). See, for example, Medzhitov, Nature. 2008 July 24; 454 (7203): 428-35. Proinflammatory agents described herein include at least two overlapping categories: 1) agents or therapies 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 tissues or activating any of these cells, or inhibiting regulatory / suppressive immune cells (e.g., regulatory T cells or MDSCs)), and 2) agents or therapies that can cause cancer cell damage (e.g., cancer cell necrosis).

[0177] In some embodiments, the proinflammatory agent triggers a proinflammatory signal on macrophages. See, e.g., Figure 5AIn 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. Proinflammatory agents can activate proinflammatory signals on macrophages directly or indirectly. For example, when used with an SHP-1 inhibitor, both TLR agonists that directly activate TLRs on macrophages, or radiation therapy that indirectly activates proinflammatory signals on macrophages, exhibit significant antitumor effects. See the Examples.

[0178] Exemplary proinflammatory agents include TLR agonists, STING activators, radiotherapy, PAMP / DAMP activators, checkpoint inhibitors, proinflammatory cytokines or chemokines, chemotherapy, bacterial components, cancer vaccines and oncolytic viruses. Other exemplary proinflammatory agents include acoustic wave therapy (such as high-intensity focused ultrasound), magnetic therapy, electrotherapy and electrostatic therapy that can kill cancer cells. See, for example, Naud et al., Nanoscale Adv., 2020, 2, 3632-3655; Rominiyi et al., Br J Cancer. February 2021; 124 (4): 697-709; Zandi et al., Cancer Med. November 2021; 10 (21): 7475-7491.

[0179] In some embodiments, the proinflammatory 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 proinflammatory cytokine or chemokine, chemotherapy, a bacterial component, a cancer vaccine, an oncolytic virus, acoustic wave therapy (e.g., high-intensity focused ultrasound), magnetic therapy, electrical therapy, and electrostatic therapy.

[0180] In some embodiments, the pro-inflammatory agent comprises an agent selected from the group consisting of a TLR agonist, a STING activator, a PAMP / DAMP activator, a pro-inflammatory cytokine or chemokine, a bacterial component, a cancer vaccine, acoustic wave therapy (e.g., high-intensity focused ultrasound), magnetic therapy, electrical therapy, and electrostatic therapy.

[0181] In some embodiments, the pro-inflammatory agent is an acoustic wave therapy (e.g., high-intensity focused ultrasound (HIFU), e.g., low-intensity focused ultrasound (LIPUS)). See, e.g., Wood et al., Ultrasound Med Biol. 2015 Apr; 41(4): 905-928; Sengupta et al., JAdv Res. 2018 Nov; 14: 97-111.

[0182] In some embodiments, the proinflammatory agent is magnetic therapy (e.g., a pulsed magnetic field, such as a static magnetic field). 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.

[0183] In some embodiments, the proinflammatory agent is electrotherapy or electrochemotherapy. See, for example, Ciria et al., Chin J Cancer Res. 2013 Apr; 25(2): 223-234; Das et al., Front Bioeng Biotechnol. 2021; 9: 795300.

[0184] In some embodiments, the proinflammatory agent is an electrostatic therapy. See, e.g., Zandi et al., Cancer Med. 2021 Nov; 10(21):7475-7491.

[0185] In some embodiments, the pro-inflammatory agent is thermoacoustic therapy. See, for example, Wen et al., Theranostics. 2017; 7(7): 1976-1989.

[0186] In some embodiments, the pro-inflammatory agent comprises a microorganism (e.g., a fragment or lysate of a microorganism). Examples of microorganisms include bacteria, fungi, and viruses.

[0187] In some embodiments, the proinflammatory agent comprises a TLR agonist (eg, R848) and a cytokine (eg, IFN-γ).

[0188] TLR agonists

[0189] In some embodiments, the proinflammatory agent comprises or is a TLR agonist.

[0190] TLRs play a crucial role in activating immune responses. TLRs recognize conserved pathogen-associated molecular patterns (PAMPs) expressed on a wide range of microorganisms, as well as endogenous DAMPs released from stressed or dying cells. TLR1, -2, -4, -5, -6, and -10 are expressed on the cell surface, while TLR3, -7, -8, and -9 are located on the endosomal membrane within the cell. TLR1 and TLR2 can heterodimerize to recognize a variety of bacterial lipid structures and cell wall components, such as triacylated lipoproteins, lipoteichoic acid, and β-glucan. TLR2 also heterodimerizes with TLR6 to bind diacylated lipopeptides. In addition, TLR2 can bind to a variety of endogenous DAMPs, such as HSPs, HMGB1, uric acid, fibronectin, and other extracellular matrix proteins. It has also been shown 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 PolyI:C. TLR4 binds LPS in a complex with the lipid A binding protein CD14 and myeloid differentiation protein 2 (MD2), as well as recognizing a variety of DAMPs. Endogenous TLR4 ligands that have been described include β-defensin 2, fibronectin extra domain A EDA, HMGB1, snapin, and tenascin C. TLR5 recognizes bacterial flagellin, TLR7 and TLR8 bind viral ssRNA, and TLR9 interacts with unmethylated CpG DNA from bacteria and some viruses. Other TLRs have recently been identified in mice based on sequence homology to the highly conserved TIR domains. TLR10 is a surface receptor whose natural ligand remains unknown. TLR11, -12, and -13 are present in mice but not in humans. TLR11 has been shown to bind to Toxoplasma gondii profibrillarin and uropathogenic Escherichia coli. The ligand for TLR12 has not yet been identified, whereas TLR13 is an endosomal receptor that recognizes VSV. See, for example, Kaczanowska et al., J Leukoc Biol. 2013 Jun; 93(6):847-63.

[0191] TLR signaling can be used as a double-edged sword in cancer. It was found that TLR stimulation of cancer cells can lead to tumor progression or suppression. For example, it was found that stimulation of TLR 2, 4, and 7 / 8 causes tumor progression via the production of immunosuppressive cytokines, increased cell proliferation, and resistance to apoptosis. R848-stimulation of pancreatic cancer cell lines that overexpress TLR7 / 8 causes increased cell proliferation and reduced chemosensitivity. On the other hand, stimulation of TLR 2, 3, 4, 5, 7 / 8, and 9 is often combined with chemotherapy or immunotherapy and can lead to tumor suppression via different pathways. See, for example, Grimmig et al., Int J Oncol. (2015) 47: 857-66; Urban-Wojciuk et al., Front Immunol. 2019; 10: 2388.

[0192] In some embodiments, the TLR agonist activates any one of the TLRs.

[0193] In some embodiments, the TLR agonist activates TLR1 or TLR2, optionally wherein the TLR agonist comprises triacylated lipoprotein, peptidoglycan, zymosan, and / or Pam3CSK4.

[0194] In some embodiments, the TLR agonist activates any one of TLR2, TLR3, TLR4, TLR5, and TLR6, optionally wherein the TLR agonist comprises a diacylated lipopeptide, a heat shock protein, HMGB1, uric acid, fibronectin, and / or an ECM protein.

[0195] In some embodiments, the TLR agonist activates TLR2, optionally wherein the TLR agonist comprises Pam3Cys, SMP-105, and / or CBLB612.

[0196] 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.

[0197] 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.

[0198] In some embodiments, the TLR agonist activates TLR5, optionally wherein the TLR agonist comprises flagellin, CBLB502, and / or M-VM3.

[0199] In some embodiments, the TLR agonist activates TLR6.

[0200] 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.

[0201] In some embodiments, the TLR agonist activates TLR7, optionally wherein the TLR agonist comprises a bistriazole group and / or R848.

[0202] In some embodiments, the TLR agonist activates TLR8, optionally wherein the TLR agonist comprises VTX1463 and / or R848.

[0203] 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.

[0204] In some embodiments, the TLR agonist activates TLR10, optionally wherein the TLR agonist comprises Pam3CSK4.

[0205] In some embodiments, the TLR agonist activates TLR11, optionally wherein the TLR agonist comprises Toxoplasma gondii profibrillar protein.

[0206] In some embodiments, the TLR agonist activates TLR12.

[0207] In some embodiments, the TLR agonist activates TLR13, optionally wherein the TLR agonist comprises VSV.

[0208] In some embodiments, the TLR agonist activates a TLR on macrophages.

[0209] In some embodiments, the TLR agonist activates TLR1, TLR2, TLR3, TLR4, TLR7, TLR8, and / or TLR9.

[0210] In some embodiments, the TLR comprises TLR1, TLR4, and / or TLR9. In some embodiments, the TLR comprises TLR9.

[0211] In some embodiments, the TLR comprises TLR2, TLR4, TLR7, and / or TLR8.

[0212] In some embodiments, the TLR agonist comprises CpG. In some embodiments, the TLR agonist comprises polyI:C. In some embodiments, the TLR agonist comprises CpG and / or polyI:C. In some embodiments, the TLR agonist comprises CpG, polyI:C and / or R848.

[0213] In some embodiments, the TLR agonist is R848, 3M-852A, motomod, bropirimine, or visammod. In some embodiments, the TLR agonist is R848.

[0214] In some embodiments, the methods described herein further comprise assessing whether the individual has a persistent infection. In some embodiments, when the individual has a persistent infection, a reduced amount of a TLR agonist is administered. In some embodiments, when the individual has a persistent infection, administration of a TLR agonist may be avoided.

[0215] Radiation therapy

[0216] In some embodiments, the proinflammatory agent comprises or is radiation therapy. Radiation activates an interconnected network of cytokines, adhesion molecules, ROS / RNS, and DAMPs, leading to a self-amplifying cascade that creates a pro-inflammatory, pro-oxidant tumor microenvironment and ultimately leads to tumor cell death. See, for example, McKelvey et al., Mamm Genome. 2018; 29(11): 843-865.

[0217] In some embodiments, radiation therapy comprises irradiating the site of the cancer to be treated.

[0218] In some embodiments, radiation therapy comprises irradiating a site different from the site of the cancer being treated.

[0219] In some embodiments, the radiation therapy is intraoperative radiation therapy ("IORT"). In certain embodiments, the radiation is localized to the tumor site. The patient may undergo intraoperative radiation before or after removal of the tumor. The tumor site may contain different types of cells, including cancerous and benign cells. In certain embodiments, the radiation therapy is stereotactic body radiation therapy ("SBRT") or stereotactic radiosurgery ("SRS").

[0220] In some embodiments, the radiation is ionizing radiation, such as particle beam radiation. Particle beam radiation can be selected from any of the following: electrons, protons, neutrons, heavy ions (such as carbon ions), or muons. Ionizing radiation can be selected from x-rays, UV light, gamma rays, or microwaves. In some embodiments, radiotherapy can comprise subjecting the patient to one or more types of radiotherapy.

[0221] In some embodiments, radiosensitizers are used to sensitize tumor cells to radiation. The use of such drugs (called radiosensitizers) provides a method for increasing the radiosensitivity of tumors to radiotherapy, thereby avoiding the need to increase the radiation dose to a level that is harmful to surrounding organs and tissues. See, for example, US9656098B2.

[0222] In some embodiments, the dose of radiation therapy is non-ablative and insufficient to eliminate the tumor (kill all tumor cells). In some embodiments, radiation therapy is selected from the group consisting of external beam radiation therapy, internal radiation therapy (brachytherapy), intraoperative radiation therapy (IORT), systemic radiation therapy, radioimmunotherapy, and administration of radiosensitizers and radioprotectants.

[0223] In some embodiments, the radiation therapy is external beam radiation therapy, which optionally includes 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).

[0224] In some embodiments, radiation therapy includes administering a radiopharmaceutical. The radiopharmaceutical can be delivered via any vehicle (e.g., 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).

[0225] In some embodiments, the radiation therapy is brachytherapy, which optionally includes interstitial brachytherapy, intracavitary brachytherapy, intraluminal radiation therapy, and intravenously administered radiolabeled molecules.

[0226] STING activators

[0227] In some embodiments, the proinflammatory agent comprises or is a STING activator.

[0228] Stimulator of IFN genes (STING, also known as TMEM173, MITA, MPYS or ERIS) is a pattern recognition receptor (PRR) that recognizes cytoplasmic 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 cytoplasmic DNA binding, cGAS converts ATP and GTP into the metazoan-specific CDN 2'3'-cGAMP for STING recognition and activation. STING is a transmembrane protein that exists as a dimer anchored in the endoplasmic reticulum membrane and forms a V-shaped pocket that enables cytoplasmic CDN binding. Ligand binding results in a significant conformational change in the C-terminal domain of STING, mediating its trafficking to the Golgi compartment. At the Golgi, STING recruits TANK-binding kinase 1 (TBK1), which promotes IRF3 phosphorylation, nuclear translocation, and potent induction of type I IFN transcription (e.g., IFN-β). STING also triggers a robust pro-inflammatory cytokine response [e.g., tumor necrosis factor (TNF)] by activating nuclear factor-κB (NF-κB), and this part of the pathway may be mediated independently of TBK1 via the closely related homolog protein IKKε. See, e.g., Peng et al., Front Immunol. 2022 Feb 25; 13: 794776; Amougezar et al., Cancers (Basel). 2021 May 30; 13(11): 2695.

[0229] In some embodiments, the STING activator is cyclic-guanosine monophosphate-adenosine monophosphate (cGAMP, eg, 3'3' cGAMP, eg, 2'3' cGAMP).

[0230] In some embodiments, the STING activator is a bacterial vector (eg, SYNB1891, STACT-TREX-1).

[0231] In some embodiments, the STING activator is a CDN compound (eg, ADU-S100, BI-STING, BMS-986301, GSK532, JNJ-4412, MK-1454, SB11285, 3'3'-cyclic AIMP).

[0232] In some embodiments, the STING activator is a non-CDN small molecule (eg, ALG-031048, E7755, JNJ-'6196, MK-2118, MSA-1, MSA-2, SNX281, SR-717, TAK676, TTI-10001).

[0233] In some embodiments, the STING activator is a nanovaccine (eg, PC7A NP, cCAMP-NP, ONM-500).

[0234] In some embodiments, the STING activator is an antibody-drug conjugate (eg, XMT-2056, CRD-5500).

[0235] Other exemplary STING activators can be found in Amougezar et al., Cancers (Basel). 2021 May 30;13(11):2695, which is incorporated herein by reference in its entirety.

[0236] PAMP / DAMP Activators

[0237] In some embodiments, the proinflammatory agent comprises or is a PAMP / DAMP activator.

[0238] Organisms sense microbial infections via innate receptors encoded in the genome, which are called pattern recognition receptors, including Toll-like receptors (TLRs), nucleotide and oligomerization domain (NOD)-like receptors, and retinoic acid-induced gene 1 (RIG-I)-like receptors. These receptors recognize pathogen-associated molecular patterns (PAMPs) expressed by bacteria, fungi, and viruses, and also bind to damage-associated molecular patterns (DAMPs) that are molecules released by sterile injury. Therefore, PAMPs and DAMPs bound to the same type of receptors initiate the same intracellular pathway and end with the same effector function. See, for example, Alisi et al., Hepatology. 2011 November; 54(5): 1500-2.

[0239] In some embodiments, the proinflammatory agent is a PAMP activator. Exemplary PAMP activators include triacyl lipopeptides, LPS, lipoproteins, peptidoglycan, zymosan, lipoteichoic acid, trypanosomal phospholipids, Pam3Cys porins, lipoarabinomannan, double-stranded RNA, poly (I: C), trypanosomal 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 bacteria, and Toxoplasma gondii LPS.

[0240] In some embodiments, the proinflammatory agent is a DAMP activator. Exemplary DAMP activators include defensins, HSP60, HSP70, messenger RNA, low molecular weight hyaluronic acid, fibrinogen, fibronectin, fx1-defensin, heparan sulfate, HSP60, HSP70, HSP90, HMGB1, and unmethylated CpG DNA.

[0241] chemotherapeutic agents

[0242] In some embodiments, the proinflammatory agent comprises or is a chemotherapeutic agent.

[0243] In some embodiments, the chemotherapeutic agent is an alkylating agent. Exemplary alkylating agents include nitrogen mustards (e.g., endomustine, 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 ethyleneimines (e.g., thiotepa).

[0244] In some embodiments, the chemotherapeutic agent is an antimetabolite. Exemplary antimetabolites include 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)).

[0245] In some embodiments, the chemotherapeutic agent is an anti-microtubule agent. Exemplary anti-microtubule agents include topoisomerase II inhibitors (e.g., anthracyclines, doxorubicin, daunomycin, edamicin, 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, daunomycin).

[0246] Other exemplary chemotherapeutic agents include hydroxyurea, tretinoin, arsenic trioxide, and proteasome inhibitors (eg, bortezomib).

[0247] Proinflammatory cytokines

[0248] In some embodiments, the proinflammatory agent is a proinflammatory cytokine.

[0249] In some embodiments, the proinflammatory cytokine promotes M1 macrophages. See, for example, Duque et al., Front Immunol. 2014; 5:491. In some embodiments, the proinflammatory cytokine comprises or is TNF, IFNγ, and / or GM-CSF.

[0250] In some embodiments, the proinflammatory cytokines comprise IL-6, TNFα, cytokines from the IL-1 family (eg, IL-1α, IL-1β, IL-18, IL-33, and IL-36), and / or IFNγ.

[0251] In some embodiments, the proinflammatory cytokine comprises a cytokine from the IL-1 family. In some embodiments, the proinflammatory cytokine comprises any one or more of IL-1α, IL-1β, 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).

[0252] Checkpoint inhibitors

[0253] In some embodiments, proinflammatory agents are checkpoint inhibitors. Immune checkpoints are pathways with inhibitory or stimulatory characteristics that maintain self-tolerance and assist immune responses. 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, for example, Marin-Acevedo et al., J Hematol Oncol 14, 45 (2021).

[0254] In some embodiments, the checkpoint inhibitor targets CTLA-4, PD-1, or PD-L1 (e.g., an antibody targeting CTLA-4, PD-1, or PD-L1).

[0255] 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 / SIRPα, CSF-1 (M-CSF) / CSF-1R, IL-1 / IL-1R3 (IL-1RAP), IL-8, SEMA4D, Ang-2, CLEVER-1, Axl, or phosphatidylserine.

[0256] In some embodiments, the checkpoint inhibitor comprises or is: ripimumab, cemiplizumab, nivolumab, pembrolizumab, atezolizumab, avelumab, durvalumab, LAG525 (IMP701), REGN3767, BI 754,091, tepolizumab (MGD013), efamod alfa (IMP321), FS118, MBG453, Sym023, TSR-022, MGC018, FPA150, EOS100850, AB928, CPI-006, monalizumab, COM701, CM24, NEO-201, difatinib, PF-04136309, MSC-1, Hu5F9-G4 (5F9), A LX148, TTI-662, RRx-001, laranolol (MCS110), LY3022855, SNDX-6352, emtansuzumab (RG7155), piritinib (PLX3397), CAN04, canakinumab (ACZ885), BMS-986253, pebinetuzumab (VX15 / 2503), teberanib, FP-1305, empatuzumab vedotin (EnaV), or bavituximab.

[0257] cancer vaccines

[0258] In some embodiments, the proinflammatory agent comprises or is a cancer vaccine. Cancer vaccines stimulate anti-tumor immunity with tumor antigens and can be delivered in the form of whole cells, peptides, nucleic acids, etc. An ideal cancer vaccine can overcome immunosuppression in tumors and induce both humoral and cellular immunity.

[0259] 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, for example, Liu et al., J Hematol Oncol 15, 28 (2022).

[0260] Cell-based vaccines were initially in the form of cancer vaccines. Cell-based cancer vaccines are usually prepared from whole cells or cell fragments, containing almost all tumor antigens and inducing a wider antigen immune response. DC vaccines are an important branch of cell-based vaccines. Personalized DC-based neoantigen cancer vaccines have shown promising anti-tumor effects in the clinic. Viruses are naturally immunogenic and their genetic material can be engineered to contain sequences encoding tumor antigens. Several recombinant viruses (such as adenoviruses) can be used as vectors to infect immune cells. 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 dissolve tumors, release tumor antigens, further increase the effectiveness of the vaccine, and produce long-term immune memory.

[0261] Peptide-based subunit vaccines (including chemical and biosynthetic preparations of predicted or known specific tumor antigens) induce robust immune responses against specific tumor antigen sites. Peptide-based subunit vaccines, when combined with adjuvants, can effectively stimulate humoral immune responses and are suitable for the prevention and treatment of viral infectious diseases.

[0262] HBV and HPV vaccines for liver and cervical cancer are primarily peptide-based subunit vaccines. In particular, virus-like particle (VLP)-based subunit vaccines that can activate cellular immune responses have shown promising antitumor activity in recent years.

[0263] Nucleic acid vaccines induce potent MHCI-mediated CD8+ T cell responses; therefore, they are a desirable cancer vaccine platform

[63] . Nucleic acid vaccines can deliver multiple antigens simultaneously to trigger humoral and cellular immunity. In addition, nucleic acid vaccines can encode full-length tumor antigens, allowing APCs to cross-present multiple epitopes or present several antigens simultaneously. Finally, nucleic acid vaccine formulation is simple and rapid, which is suitable for the development of personalized neoantigen cancer vaccines.

[0264] Oncolytic viruses

[0265] In some embodiments, the proinflammatory agent is an oncolytic virus (OV). Oncolytic viruses (OV) are organisms that can identify, infect, and dissolve different cells in the tumor environment, aiming to stabilize and reduce tumor progression. They can present natural tropism to cancer cells or be genetically directed to identify specific targets. See, for example, Apolonio et al., World J Virol. September 25, 2021; 10(5): 229-255.

[0266] Oncolytic viruses represent an exciting new approach to cancer therapy. These viruses have a remarkable ability to seek out and kill cancer cells while leaving healthy cells unharmed and enhancing the immune system's ability to recognize and kill cancer cells. See, for example, Cancer Cell. 2022 Aug 15; S1535-6108(22)00357-9.

[0267] In some embodiments, the oncolytic virus comprises or is an adenovirus (e.g., ONYX-15, LOAd703 virus), a proparvovirus, a parvovirus (e.g., H-1PV), a vaccinia virus (VACV), a reovirus (e.g., Relaison), or a herpes simplex virus (HSV, e.g., HSV-1, HSV-2, G207, L1BR1, HF10, T-VEC, Orien X010).

[0268] Other exemplary oncolytic viruses include JX-593, Coxsackievirus A21 (CVA21), Maraba virus or its MG1 variant, DNX2440 adenovirus, fowlpox virus, and Sendai virus.

[0269] cell

[0270] In some embodiments, the proinflammatory 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., 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 neutrophil cell expressing a CAR). 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.

[0271] Immune cells, monocytes, or macrophages

[0272] The immune cells described herein encompass multiple types of immune cells.

[0273] In some embodiments, the immune cells comprise monocytes or macrophages as described herein. In some embodiments, macrophages are identified by F4 / 80 expression. In some embodiments, the macrophages have an M1 phenotype. In some embodiments, at least 50%, 60%, 70%, 80%, 90%, 95%, 98% or 99% of the macrophages in the immune cells have an M1 phenotype.

[0274] In some embodiments, the macrophages are engineered to be defective in SHP-1 expression and / or activation. In some embodiments, the monocytes or macrophages express reduced levels of SHP-1 for at least a period of time (e.g., for at least 1, 2, 3, 4, or 5 days), or are resistant to activation for at least a period of time (e.g., for at least 1, 2, 3, 4, or 5 days). In some embodiments, the period of time is no more than about 10, 9, 8, 7, 6, 5, 4, or 3 days.

[0275] In some embodiments, SHP-1 activity in monocytes or macrophages is reduced for no more than about 5 consecutive days (eg, no more than 5, 4, or 3 days) before the level of SHP-1 activity returns to normal.

[0276] Methods for engineering monocytes or macrophages to transiently express reduced levels of SHP-1 are well known in the art. Exemplary methods include contacting monocytes or macrophages with a SHP-1 inhibitor (e.g., 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 or activates SHP-1)) described herein in vivo or in vitro.

[0277] In some embodiments, the immune cells comprise T cells (e.g., CAR-T cells).

[0278] In some embodiments, the immune cells comprise NK cells (eg, CAR-NK cells).

[0279] In some embodiments, the immune cell comprises a neutrophil (e.g., a CAR-expressing neutrophil cell).

[0280] In some embodiments, the immune cells comprise antigen presenting cells (APCs, such as dendritic cells).

[0281] In some embodiments, the immune cells are derived from the same individual (ie, autologous). In some embodiments, the immune cells are allogeneic.

[0282] In some embodiments, the immune cells are engineered to express a chimeric antigen receptor, optionally wherein the chimeric antigen receptor specifically binds to a tumor antigen.

[0283] In some embodiments, the immune cells express high levels of MHC-I, MHC-II, CD80, and / or CD86. In some embodiments, the immune cells express high levels of MHC-I, MHC-II, CD80, and / or CD86 when the expression levels of MHC-I, MHC-II, CD80, and / or CD86 on the immune cells are comparable (e.g., at least 50% greater) to those on activated antigen presenting cells (APCs).

[0284] In some embodiments, the immune cells express one or more proinflammatory cytokines, optionally wherein the one or more proinflammatory cytokines comprise TNFα and / or IL-12.

[0285] In some embodiments, the immune cells do not express significant levels of TGFβ and / or IL-10.

[0286] In some embodiments, the SHP-1 inhibitor and the immune cells 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 SHP-1 inhibitor and the immune cells are administered within 4 hours of each other.

[0287] In some embodiments, the SHP-1 inhibitor, immune cells, and proinflammatory agent described above 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 cells are administered simultaneously or concurrently with the SHP-1 inhibitor and / or proinflammatory agent.

[0288] Inflammatory response or ongoing infection

[0289] There is a lot of evidence that both acute and chronic inflammation are associated with the development and progression of cancer. Research progress on inflammation reveals the relationship between inflammatory processes and neoplastic transformation, the progression of tumors, and the development of metastasis and recurrence. In addition, tumor aggressiveness procedures (both surgery and biopsy) affect the remaining tumor cells by increasing their survival, proliferation, and migration. One of the concepts to explain this phenomenon is to induce wound healing responses. Although it is necessary for tissue repair in normal tissues, in tumor tissues, inducing acquired and innate immune responses related to wound healing stimulates tumor cell survival, angiogenesis, and circulating tumor cell extravasation. See, for example, Singh et al., Ann Afr Med. 2019 July-September; 18(3): 121-126; Piotrowski et al., Rep Pract Oncol Radiother. 2020 May-June; 25(3): 422-427.

[0290] However, as demonstrated in this application, the combined use of SHP-1 inhibitors and proinflammatory agents unleashes a proinflammatory response and transforms the immunosuppressive tumor environment into one with hallmarks of inflammation. See, e.g. Figure 7F These results provide support for the use of the methods described herein to treat individuals experiencing inflammatory responses.

[0291] 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, for example, by a) an increase in one or more (e.g., at least one, two, three, four, five) inflammatory cytokines (e.g., IFNγ, IL-12b, TNFα, 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 (e.g., TGFb1, TGFb2, TGFb3), c) an increase in infiltrating immune cells (e.g., T cells, NK cells, macrophages, neutrophils), d) a decrease in suppressive immune cells (e.g., MDSCs), and / or e) an increase in one or more (e.g., at least one, two, three, four, or five) immunogenic co-stimulatory molecules (e.g., CD80, CD86, OX40L, CD40, ICOS-L, PD-L1, GITRL) in tissue (e.g., tumor tissue) or immune cells (e.g., macrophages).

[0292] In some embodiments, the inflammatory response is an acute inflammatory response.

[0293] In some embodiments, the inflammatory response is located in the tumor. In some embodiments, the inflammatory response is located at a site different from the tumor.

[0294] In some embodiments, an inflammatory response is present 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 (e.g., IFNγ, IL-12b, TNFα, 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 (e.g., TGFb1, TGFb2, TGFb3), c) an increase in infiltrating immune cells (e.g., T cells, NK cells, macrophages, neutrophils), d) a decrease in suppressive immune cells (e.g., MDSCs), and / or e) an increase in one or more (e.g., at least one, two, three, four, or five) immunogenic co-stimulatory molecules (e.g., CD80, CD86, OX40L, CD40, ICOS-L, PD-L1, GITRL) in tissue (e.g., tumor tissue) or immune cells (e.g., macrophages).

[0295] In some embodiments, the increase described herein refers to an amount that is at least about 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175% or 200% higher than a reference state, optionally wherein the reference state is when the individual is neither treated with the methods described herein nor infected with a pathogen. In some embodiments, the increase described herein refers to an amount that 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 state, optionally wherein the reference state is when an individual is neither treated with the methods described herein nor infected with a pathogen. In some embodiments, the reference state is when a healthy individual is not infected with a pathogen.

[0296] In some embodiments, a reduction as described herein refers to an amount that is at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 99.9% lower than a reference state, optionally wherein the reference state is when the individual is neither treated with the methods described herein nor infected with a pathogen. In some embodiments, the reference state is when a healthy individual is not infected with a pathogen.

[0297] In some embodiments, the subject has an inflammatory response (eg, in a tumor, eg, in a site different from the tumor) within about one week, six days, five days, four days, three days, two days, or one day before and / or after administration of the SHP-1 inhibitor.

[0298] In some embodiments, the individual has an ongoing inflammatory response (eg, in a tumor, eg, in a site different from the tumor) when the SHP-1 inhibitor is administered.

[0299] In some embodiments, the individual has an ongoing infection when the SHP-1 inhibitor is administered.In some embodiments, the method further comprises assessing the individual for the presence of an infection, such as an infection associated with a virus, fungus, and / or bacteria.

[0300] In some embodiments, the individual has an ongoing infection (eg, a bacterial infection, a viral infection, a fungal infection), and the method further comprises administering an antibacterial therapy (eg, an antibiotic), an antiviral therapy, an antimicrobial therapy, or an antiprotozoal therapy.

[0301] Immunogenic cell death

[0302] In some embodiments, the individual has immunogenic cell death when treated with the methods described herein.

[0303] Immunogenic 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 chemotherapeutic agents 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 poly ADP-ribose polymerase (PARP) inhibitors; and (4) various physical modalities, covering hypericin-based and redaporfin-based photodynamic therapy, extracorporeal photochemotherapy, various forms of ionizing radiation, high hydrostatic pressure, and severe heat shock. It involves the activation of the immune system against cancer in an immunocompetent host. ICD involves the release of damage-associated molecular patterns (DAMPs) from dying tumor cells, which lead to the activation of tumor-specific immune responses, thereby resulting in long-term efficacy of anticancer drugs through a combination of direct cancer cell killing and 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 box-1 (HMGB1), type I IFN and members of the IL-1 cytokine family. See, for example, Ahmed et al., Mol Oncol. 2020 December; 14(12): 2994-3006 and Fucikova et al., Cell Death Dis. 2020 November 26; 11(11): 1013.

[0304] Key DAMPs of cell death that are perceived as immunogenic include calreticulin, high mobility group box 1 (HMGB1), ATP, annexin A1 (ANXA1), and type I IFN. The main markers of immunogenic cell death (ICD) can be assessed by flow cytometry, (immuno)fluorescence microscopy, immunoblotting, or luminescence assays based on a variety of different approaches. See, for example, Cell Death Dis. 2020 Nov 26; 11(11): 1013.

[0305] In some embodiments, the individual has an ICD (eg, in a tumor, eg, in a site different from the tumor) within about one week, six days, five days, four days, three days, two days, or one day before and / or after administration of the SHP-1 inhibitor.

[0306] In some embodiments, the individual has persistent ICD (eg, in the tumor, eg, in a site different from the tumor) when the SHP-1 inhibitor is administered.

[0307] In some embodiments, a subject has ICD when a sample from a cancer has higher levels of one or more (e.g., at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%) DAMPs than a reference sample (e.g., a corresponding sample from a healthy control, e.g., a sample from a cancer before administration of a therapy that induces ICD). In some embodiments, the DAMPs are selected from the group consisting of: endoplasmic reticulum (ER) chaperone proteins (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 metabolite ATP, and type I interferon (IFN).

[0308] individual

[0309] In some embodiments, the individual has a solid tumor. In some embodiments, the individual has a hematological cancer.

[0310] In some embodiments, the subject has advanced cancer. In some embodiments, the subject has late stage cancer. In some embodiments, the subject has a malignant cancer. In some embodiments, the subject has stage II, III, or IV cancer. In some embodiments, the subject has an inoperable tumor and / or metastasis. In some embodiments, the subject is terminally ill.

[0311] In some embodiments, the subject has received (e.g., within 1, 2, 4, 8, 12, 16, 20, or 24 hours, e.g., within 1, 2, 3, 4, 5, 6, or 7 days before administration of the SHP-1 inhibitor) a therapy that induces an inflammatory response or immunogenic cell death (e.g., radiation therapy). In some embodiments, the subject is about to receive (e.g., within 1, 2, 4, 8, 12, 16, 20, or 24 hours, e.g., within 1, 2, 3, 4, 5, 6, or 7 days after administration of the SHP-1 inhibitor) a therapy that induces an inflammatory response or immunogenic cell death (e.g., radiation therapy).

[0312] In some embodiments, the subject has received (e.g., within 1, 2, 4, 8, 12, 16, 20, or 24 hours, e.g., within 1, 2, 3, 4, 5, 6, or 7 days prior to administration of a SHP-1 inhibitor) a proinflammatory agent (e.g., any of the proinflammatory agents described herein). In some embodiments, the subject is about to receive (e.g., within 1, 2, 4, 8, 12, 16, 20, or 24 hours, e.g., within 1, 2, 3, 4, 5, 6, or 7 days after administration of a SHP-1 inhibitor) a proinflammatory agent (e.g., any of the proinflammatory agents described herein).

[0313] In some embodiments, the individual does not have an autoimmune disease.

[0314] In some embodiments, the individual is female. In some embodiments, the individual is male.

[0315] In some embodiments, the individual is a human. In some embodiments, the individual is at least about 50, 55, 60, 65, 70, or 75 years old.

[0316] In some embodiments, an individual is selected for treatment based on high expression levels and / or high activation levels of SHP-1 in tumor tissue. In some embodiments, an individual has high expression levels and / or high activation levels of SHP-1 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% greater than a reference expression level and / or reference activation level of SHP-1. In some embodiments, a subject has a high expression level and / or a high activation level of SHP-1 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 greater than a reference expression level and / or reference activation level of SHP-1. In some embodiments, the reference expression level or reference activation level of SHP-1 is the corresponding expression or activation level of SHP-1 in a reference state in which the subject is not treated with a proinflammatory agent (or any immunotherapy).

[0317] 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 before administering an agent that reduces systemic inflammation. Cytokine release syndrome can damage most organ systems or cause organ failure in most organ systems. For example, organs that may become damaged due to CRS may include, but are not limited to, lungs, kidneys, liver, brain, heart, spleen, or any combination thereof, such as multiple organ failure.

[0318] In some embodiments, an agent that reduces systemic inflammation is administered to an individual. In some embodiments, administration occurs before the individual develops 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 may include fever, fatigue, headache, rash, arthralgia, and myalgia. Mild CRS can be treated by treating the symptoms or by administering anti-inflammatory drugs (e.g., corticosteroids). Mild CRS typically resolves within one to two weeks and does not require or require hospitalization.

[0319] In some embodiments, the individual does not suffer from severe cytokine release syndrome. In some embodiments, the individual does not suffer from grade 2 CRS. In some embodiments, the individual does not suffer from grade 3 CRS. In some embodiments, the individual does not suffer from grade 4 CRS. More serious cases are characterized by hypotension and hyperthermia, and severe CRS can progress to uncontrolled systemic inflammatory response with circulatory shock, vascular leak, disseminated intravascular coagulation, and multi-organ system failure requiring vasopressors. More serious cases of CRS typically require hospitalization. Common laboratory abnormalities in patients with CRS include cytopenia, elevated creatinine and liver enzymes, deranged coagulation parameters, and high CRP. There are four current grading systems for cytokine release syndrome, as shown in Table 1 below. 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 incorporated herein by reference in their entireties.

[0320] In some embodiments, prior to administering an agent that reduces systemic inflammation, the individual has developed CRS. 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, an agent that reduces systemic inflammation is administered to an individual who has developed CRS. In some embodiments, agents that reduce systemic inflammation improve, eliminate, or reverse CRS, including organ damage, such as proinflammatory organ damage (e.g., nephritis, hepatitis, pneumonia, myocarditis, appendicitis).

[0321] Table 1. Medical grading system for cytokine release syndrome.

[0322]

[0323]

[0324] 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 storms appear to be primarily the result of nonspecific T cell activation, while CRS is more commonly a direct result of antigen-specific T cell activation. The clinical manifestations of cytokine storms and CRS can be similar (Liu, D. and Zhao, J., J Hematol Oncol. 2018 September 24; 11 (1): 121).

[0325] cancer

[0326] The cancer described herein can be of any type or species. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a hematological cancer.

[0327] In some embodiments, the cancer is advanced cancer. In some embodiments, the cancer is advanced cancer. In some embodiments, the cancer is terminal cancer. In some embodiments, the cancer is in stage II, III, or IV. In some embodiments, the cancer is an inoperable tumor and / or is malignant.

[0328] In some embodiments, 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 in length.

[0329] Examples of cancers described herein include, but are not limited to, adrenocortical carcinoma, myeloid metaplasia of unknown cause, AIDS-related cancers (e.g., AIDS-related lymphoma), anal cancer, appendiceal cancer, astrocytomas (e.g., cerebellar and cerebral), basal cell carcinoma, bile duct cancer (e.g., extrahepatic), bladder cancer, bone cancer (osteosarcoma and malignant fibrous histiocytoma), brain tumors (e.g., gliomas, brainstem gliomas, cerebellar or cerebral astrocytomas (e.g., pilocytic astrocytomas, diffuse astrocytomas, anaplastic (malignant) astrocytomas), malignant gliomas, Ependymoma, oligodenglioma, meningioma, craniopharyngioma, hemangioblastoma, medulloblastoma, supratentorial primitive neuroectodermal tumor, optic pathway and hypothalamic glioma and glioblastoma), breast cancer, bronchial adenoma / carcinoid, carcinoid tumor (e.g., gastrointestinal carcinoid tumor), cancer of unknown primary site, central nervous system lymphoma, cervical cancer, colon cancer, colorectal cancer, chronic myeloproliferative disease, endometrial cancer (e.g., uterine cancer), ependymoma, esophageal cancer, Ewing's family of tumors (e.g., ovarian cancer), esophageal cancer, ovarian ...tumor), eye cancer (e.g., intraocular melanoma and retinoblastoma), gallbladder cancer, gastric (stomach) cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), germ cell tumors (e.g., extracranial, extragonadal, ovarian), gestational trophoblastic tumor, head and neck cancer, hepatocellular (liver) cancer (e.g., hepatocarcinoma and heptoma), hypopharyngeal cancer, islet cell cancer (endocrine pancreas), laryngeal cancer, leukemia, lip and oral cavity cancer, and oral cancer , liver cancer, lung cancer (e.g., small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma, and lung squamous cell carcinoma), lymphocytic neoplasms (e.g., lymphoma), medulloblastoma, melanoma, mesothelioma, metastatic squamous neck cancer, oral cancer, multiple endocrine neoplasia syndrome, myelodysplastic syndrome, myelodysplastic / myeloproliferative disorders, 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-grade malignant potential), pancreatic cancer, parathyroid cancer, penile cancer, peritoneal cancer, pharyngeal cancer, pheochromocytoma, pineoblastoma and supratentorial primitive neuroectodermal tumors, pituitary tumors, pleuropulmonary blastoma, lymphoma, primary central nervous system lymphoma (microglioma), pulmonary lymphangioleiomyomatosis, rectal cancer, kidney cancer, renal pelvis and ureter cancer (transitional cell carcinoma), rhabdomyosarcoma, salivary gland cancer, skin cancer (e.g., non-melanomas (e.g., squamous cell carcinoma), melanoma, and Merkel cell carcinoma), Examples of the diseases that are considered include: Merkel cell carcinoma, small bowel cancer, squamous cell carcinoma, testicular cancer, pharyngeal cancer, thymoma and thymic cancer, thyroid cancer, tuberous sclerosis, urethral cancer, vaginal cancer, vulvar cancer, Wilms' tumor, and post-transplantation lymphoproliferative disorder (PTLD), abnormal blood vessel proliferation associated with macular degeneration, edema (e.g., edema associated with brain tumors), and Meigs' syndrome.

[0330] In some embodiments, cancer is a viral infection-related cancer. In some embodiments, cancer is human papillomavirus (HPV)-related cancer (e.g., HPV-related cervical cancer, e.g., HPV-related head and neck cancer, e.g., HPV-related squamous cell carcinoma). In some embodiments, cancer is human herpesvirus 8 (HHV8)-related cancer (e.g., Kaposi's sarcoma). In some embodiments, cancer is human T-lymphotropic virus (HTLV-1)-related cancer (e.g., adult T-cell leukemia or lymphoma). In some embodiments, cancer is Epstein-Barr virus (Epstein-Barr virus; EBV)-related cancer (e.g., Burkitt lymphoma, Hodgkin's and non-Hodgkin's lymphoma, gastric cancer). In some embodiments, cancer is hepatitis B virus (HBV)-related cancer (e.g., liver cancer). In some embodiments, cancer is hepatitis C virus-related cancer (e.g., liver cancer, non-Hodgkin's lymphoma).

[0331] In some embodiments, the cancer is liver cancer, kidney cancer, endometrial cancer, thymic epithelial neoplasm, lung cancer, spindle cell sarcoma, chondrosarcoma, uterine leiomyoma, colon cancer, or pancreatic cancer.

[0332] In some embodiments, the cancer has been treated with and / or failed one or more prior therapies (e.g., immune checkpoint blockade therapy (e.g., PD-1 antibody), chemotherapy, surgery, cell therapy (e.g., allogeneic NK cell infusion therapy)).

[0333] In some embodiments, the cancer is a relapsed or refractory cancer.

[0334] In some embodiments, the cancer is refractory to one or more of radiation therapy, chemotherapy, or immunotherapy (e.g., checkpoint blockade).

[0335] Dosing, administration methods and delivery vehicles

[0336] The SHP-1 inhibitors, proinflammatory agents, and immune cells (eg, monocytes / macrophages) described herein can be administered at any desired dose. Exemplary dosing regimens are described, for example, in the "SHP-1 Inhibitors" section.

[0337] In some aspects, the size of the dose of a proinflammatory agent, SHP-1 inhibitor, and / or immune cell (e.g., monocyte / macrophage) is determined based on one or more criteria such as: the disease load in the subject, such as tumor load, mass, size or extent, extent or type of metastasis; stage; and / or the likelihood or incidence of a subject developing a toxic outcome, such as CRS, macrophage activation syndrome, tumor lysis syndrome, neurotoxicity, and / or a host immune response to the activated immune cells administered. For example, in some aspects, the number of monocytes or macrophages administered in a dose is determined based on the tumor load present in the subject prior to the start of administration of the cell dose.

[0338] The proinflammatory agent, SHP-1 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., intravenous or subcutaneous injection. In some embodiments, the proinflammatory agent, SHP-1 inhibitor, and / or monocytes or macrophages are administered systemically (e.g., intravenously, subcutaneously, or intraperitoneally). In some embodiments, the proinflammatory agent, SHP-1 inhibitor, and / or monocytes or macrophages are administered locally (e.g., intratumorally).

[0339] In some embodiments, the proinflammatory agent, SHP-1 inhibitor, and / or immune cells (e.g., monocytes / macrophages) are administered parenterally, intrapulmonary, and intranasally, and, if necessary for local treatment, intralesional or intratumoral administration. Parenteral infusion includes intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. In some embodiments, the proinflammatory agent and / or SHP-1 inhibitor is administered orally.

[0340] In some embodiments, immune cells (such as monocytes / macrophages) and proinflammatory agents are administered simultaneously. In some embodiments, monocytes or macrophages and proinflammatory agents are administered in parallel. In some embodiments, immune cells (such as monocytes / macrophages) and proinflammatory agents are administered sequentially. In some embodiments, immune cells (such as monocytes / macrophages) and proinflammatory agents are administered within about 7, 6, 5, 4, 3, 2 or 1 days. In some embodiments, immune cells (such as monocytes / macrophages) and proinflammatory agents are administered within about 24, 16, 12, 8, 4, 2 or 1 hours. In some embodiments, immune cells (such as monocytes / macrophages) and proinflammatory agents are administered within 30 minutes.

[0341] In some embodiments, the SHP-1 inhibitor and the proinflammatory agent are administered simultaneously. In some embodiments, the SHP-1 inhibitor and the proinflammatory agent are administered concurrently. In some embodiments, the SHP-1 inhibitor and the proinflammatory agent are administered sequentially. In some embodiments, the SHP-1 inhibitor and the proinflammatory agent are administered within about 7, 6, 5, 4, 3, 2, or 1 day. In some embodiments, the SHP-1 inhibitor and the proinflammatory agent are administered within about 24, 16, 12, 8, 4, 2, or 1 hour. In some embodiments, the SHP-1 inhibitor and the proinflammatory agent are administered within 30 minutes.

[0342] It is also contemplated that the SHP-1 inhibitors and / or proinflammatory agents described herein may be delivered via any appropriate vehicle or method. In some embodiments, the SHP-1 inhibitors and / or proinflammatory agents are delivered directly to tumor tissue. For this purpose, various carrier systems may be used. See, for example, 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 SHP-1 inhibitors and / or proinflammatory agents are delivered via nanoparticles. In some embodiments, the SHP-1 inhibitors and / or proinflammatory agents are delivered via a controlled release system. In some embodiments, the SHP-1 inhibitors and / or proinflammatory agents are delivered via a biomaterial implant scaffold. In some embodiments, the SHP-1 inhibitors and / or proinflammatory agents are delivered via an injectable biomaterial scaffold. In some embodiments, the SHP-1 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.

[0343] In some embodiments, the SHP-1 inhibitor and / or proinflammatory agent is delivered via cells. See, e.g., Millian et al., Ther Deliv. 2012 Jan;3(1):25-41. In some embodiments, the cells comprise macrophages. See, e.g., Visser et al., Front Pharmacol. 2019 Jan 25;10:22. In some embodiments, the cells comprise polymer-encapsulated human retinal pigment epithelial (aRPE) cells. See, e.g., Nash et al., Clin Cancer Res. 2022 Aug 22;CCR-22-1493. In some embodiments, the cells are encapsulated in a biocompatible material (e.g., a biocompatible alginate capsule, as discussed in Nash et al.).

[0344] In some embodiments, the SHP-1 inhibitor and / or proinflammatory agent is associated with an antibody construct. In some embodiments, the SHP-1 inhibitor and / or proinflammatory 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).

[0345] In some embodiments, the SHP-1 inhibitor and / or proinflammatory agent is delivered via a method or device that facilitates delivery to a specific organ (e.g., an organ with a tumor). Examples of such methods or devices are described, for example, in Alsaggar et al., J Drug Target. 2018 Jun-Jul; 26(5-6):385-397; Zhao et al., Cell. 2020 Apr 2; 181(1):151-167 (incorporated by reference in their entireties).

[0346] In an embodiment, the SHP-1 inhibitor is delivered via a controlled drug delivery system (e.g., a slow release system or vehicle, such as a sustained release system or vehicle). Examples of such systems can be found, for example, in 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 entirety.

[0347] VI. Compositions Containing SHP-1 Inhibitors

[0348] The present application also provides compositions (eg, pharmaceutical compositions) comprising a SHP-1 inhibitor, a pro-inflammatory agent, and / or immune cells for use in treatment, as described above.

[0349] In some embodiments, a composition (e.g., a pharmaceutical composition) is provided, comprising a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a proinflammatory agent (e.g., any proinflammatory agent described herein). In some embodiments, the composition further comprises immune cells (e.g., monocytes or macrophages described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier.

[0350] In some embodiments, a composition (e.g., a pharmaceutical composition) is provided, comprising a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a TLR agonist (e.g., CpG, polyI:C, and / or R848). In some embodiments, the composition further comprises immune cells (e.g., monocytes or macrophages as described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier.

[0351] In some embodiments, a composition (e.g., a pharmaceutical composition) is provided, comprising a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a STING activator (e.g., cGAMP, e.g., 2'3'-cGAMP, e.g., 3'3'-cGAMP). In some embodiments, the composition further comprises an immune cell (e.g., a monocyte or macrophage as described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier.

[0352] In some embodiments, a composition (e.g., a pharmaceutical composition) is provided that comprises a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a chemotherapeutic agent (e.g., azathioprine (AZA), such as gemcitabine). In some embodiments, the composition further comprises an immune cell (e.g., a monocyte or macrophage as described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier.

[0353] In some embodiments, a composition (e.g., a pharmaceutical composition) is provided, comprising a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a pro-inflammatory cytokine (e.g., IL-1b, IL-18, IL-6, and / or TNFα). In some embodiments, the composition further comprises immune cells (e.g., monocytes or macrophages as described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier.

[0354] In some embodiments, a composition (e.g., a pharmaceutical composition) is provided, comprising a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a checkpoint inhibitor (e.g., an anti-PD-L1 antibody, an anti-PD-1 antibody, or an anti-CLTA4 antibody). In some embodiments, the composition further comprises an immune cell (e.g., a monocyte or macrophage as described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier.

[0355] In some embodiments, a composition (e.g., a pharmaceutical composition) is provided, comprising a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a bacterial component (e.g., LPS). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier. In some embodiments, the composition further comprises an immune cell (e.g., a monocyte or macrophage as described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier.

[0356] In some embodiments, a composition (e.g., a pharmaceutical composition) is provided, comprising a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and an agent that promotes immunogenic cell death (ICD). In some embodiments, the composition further comprises immune cells (e.g., monocytes or macrophages as described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier.

[0357] In some embodiments, a composition (e.g., a pharmaceutical composition) is provided, comprising a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and an agent for use in radiation therapy (e.g., any radiation therapy described herein). In some embodiments, the composition further comprises immune cells (e.g., monocytes or macrophages described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier.

[0358] In some embodiments, a composition (e.g., a pharmaceutical composition) is provided, comprising a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a PAMP / DAMP activator (e.g., any of the PAMP / DAMP activators described herein). In some embodiments, the composition further comprises an immune cell (e.g., a monocyte or macrophage described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier.

[0359] In some embodiments, a composition (e.g., a pharmaceutical composition) is provided, comprising a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and a cancer vaccine (e.g., any cancer vaccine described herein). In some embodiments, the composition further comprises immune cells (e.g., monocytes or macrophages described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier.

[0360] In some embodiments, a composition (e.g., a pharmaceutical composition) is provided, comprising a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and an oncolytic virus (e.g., any oncolytic virus described herein). In some embodiments, the composition further comprises an immune cell (e.g., a monocyte or macrophage described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier.

[0361] In some embodiments, a composition (e.g., a pharmaceutical composition) is provided, comprising a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and an agent for use in acoustic wave therapy (e.g., any acoustic wave therapy described herein). In some embodiments, the composition further comprises immune cells (e.g., monocytes or macrophages described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier.

[0362] In some embodiments, a composition (e.g., a pharmaceutical composition) is provided, comprising a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and an agent for magnetic therapy (e.g., any magnetic therapy described herein). In some embodiments, the composition further comprises immune cells (e.g., monocytes or macrophages described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier.

[0363] In some embodiments, a composition (e.g., a pharmaceutical composition) is provided, comprising a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and an agent for use in an electrical or electrochemical treatment (e.g., any of the electrical or electrochemical treatments described herein). In some embodiments, the composition further comprises an immune cell (e.g., a monocyte or macrophage described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier.

[0364] In some embodiments, a composition (e.g., a pharmaceutical composition) is provided, comprising a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and an agent for use in electrostatic therapy (e.g., any of the electrostatic therapies described herein). In some embodiments, the composition further comprises immune cells (e.g., monocytes or macrophages described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier.

[0365] Example

[0366] The following examples are intended to be merely exemplary of the present invention, and therefore should not be construed as limiting the invention in any way.The following examples and detailed description are offered by way of illustration and not limitation.

[0367] Example 1.

[0368] SHP-1 is abundantly expressed in macrophages. Proteomic studies examining the expression of non-receptor protein tyrosine phosphatases (PTPs) in macrophages revealed that SHP-1 has the highest expression level. Figure 15 As shown, SHP-1 is the most abundant protein tyrosine phosphatase expressed in macrophages of both human and murine origin. SIRPα is an activated macrophage inhibitory receptor (iR) involved in SHP-1. Prior to analysis, human monocyte-derived macrophages were either left unstimulated (M0) or stimulated with IFNγ / LPS (M1) or IL-4 (M2) to induce an activated phenotype. (The housekeeping gene GAPDH was also assayed in parallel.)

[0369] Despite its high expression, we found that SHP-1 activity in tumor-associated macrophages remains low unless the macrophages are “surrounded / contacted” by tumor cells and simultaneously stimulated with TLR agonists or other proinflammatory activators. Figure 2A Under these conditions, inhibition of SHP-1 activity by the covalent inhibitor TPI-1 (8) reduces SHP-1-mediated protein dephosphorylation, leading to a marked enhancement of signaling mediated by TLRs and other factors.

[0370] As shown ( Figure 2C ), the phosphorylation of macrophages stimulated with IFNγ / LPS was completely inhibited and, consequently, the activation of STAT-1 (pSTAT-1) and Erk1 / 2 (pErk1 / 2) in the presence of cancer cell ligation, whereas the inhibition of SHP-1 exerted by cancer cells, released in a dose-dependent manner by TPI-1, allowed signal transduction and activation of STAT-1 (pSTAT-1) and Erk1 / 2 (pErk1 / 2). Consistently, this effect of TPI-1 on SHP-1 inhibition led to elevated proinflammatory cytokine production and expression of immunogenic antigen presentation machinery by macrophages ( Figure 2D and Figure 2E Macrophages with high SHP-1 activity in the tumor environment have reduced proinflammatory cytokine production but produce high IL-10 upon IFNγ / LPS stimulation (see Figure 2D , blue bars). This enhanced immunosuppression was reversed by SHP-1 inhibition. In addition, SHP-1 inhibition also enabled pro-inflammatory activated macrophages to overcome the "don't-eat-me" barrier and initiate potent phagocytosis toward cancer cells, regardless of whether the cancer cells expressed CD47 ( Figure 2F and Figure 2G ).

[0371] Example 2.

[0372] In addition to TPI-1, several other SHP-1 inhibitors have been tested, such as PTP inhibitor-I, PTP inhibitor-III, and recently reported vitamin E (9, 10) and Phomopsis xanthones A and B (PXA and PXB) (11). Given that PKCθ regulates SHP-1 activity via phosphorylation at Ser591 (12, 13), PKCθ inhibitors and activators have also been tested in various assays.

[0373] Figures 3A to 3E As shown in the results, among these compounds, TPI-1 exhibited the strongest effect and potently inhibited SHP-1 activity at low concentrations. Vitamin E derivatives and the PKCθ activator PMA modestly inhibited SHP-1. We also investigated the effects of inhibition of SHP-2, a close family member of SHP-1. In contrast to SHP-1 inhibition, SHP-2 inhibition did not significantly reduce PTP activity induced by proinflammatory activation and cancer cell attachment, nor did it confer phagocytosis of cancer cells by activated macrophages. These studies suggest that SHP-2 is regulated differently from SHP-1, and that SHP-1, but not SHP-2, controls macrophage proinflammatory responses, immunogenic antigen presentation, and phagocytosis of cancer cells.

[0374] Example 3.

[0375] SHP-1 was found to have a rapid turnover rate. Macrophages were treated with the covalent inhibitor TPI-1 for 15 min, followed by washing and removal of inhibitor availability. Strong SHP-1 inhibition was observed immediately after TPI-1 treatment, an effect associated with increases in pSTAT-1 and pErk1 / 2 induced by IFNγ / LPS. However, this type of pulsed SHP-1 inhibition did not persist for more than a few hours (5-8 h), as SHP-1 activity began to recover and reached over 50% within 12-16 h, thereby acquiring the ability to reduce IFNγ / LPS-induced signaling. Figure 4A Although SHP-1 activity was altered, total protein did not appear to change. Treatment of macrophages with TPI-1 without or with partial removal of the inhibitor prolonged the inhibition of SHP-1 activity ( Figure 4B ).

[0376] Example 4.

[0377] Further molecular and cellular signaling studies demonstrated that, in the tumor environment, tumor cells ligate macrophage iRs (e.g., SIRPα, LI1RB, and Siglec) via their cell surface counterreceptors (e.g., CD47, MHC, and carbohydrates), which then drive high-level SHP-1 activation via their cytoplasmic ITIMs that become tyrosine phosphorylated in the presence of proinflammatory stimuli, bind to SHP-1, and thereby disengage the SH2 domain from the auto-inhibition of SHP-1 ( Figure 5A (depicted in ). Neither extracellular ligation of iRs nor proinflammatory stimuli alone drive robust ITIM phosphorylation and, consequently, SHP-1 activation, but their simultaneous presence achieves this. The underlying mechanism suggests that extracellular ligation of iRs induces a change in their cytoplasmic structure to an "open" form, exposing ITIMs to phosphorylation, while proinflammatory signals simultaneously induce activation of Src family tyrosine kinases (TKs) to mediate ITIM phosphorylation. Figure 5B Studies have shown that SIRPα, an essential macrophage iR, requires both extracellular ligation of CD47 and macrophage stimulation with cytokines or TLR agonists for its ITIM phosphorylation.

[0378] These studies also found that macrophage iRs (e.g., SIRPα) can recruit both SHP-1 and SHP-2 via cytoplasmic ITIM phosphorylation. However, when macrophages are stimulated with activating cytokines (TNFα, IL-17A, IL-6, or IFNγ) or TLR agonists (LPS, CpG, or PolyIC), SHP-1 binding occurs only under proinflammatory conditions; whereas SHP-2 binding is dominated by immunosuppressive IL-4, IL-10, or TGFβ ( Figure 5C ).

[0379] Example 5

[0380] Similar findings were obtained in solid tumors in vivo. As tumors progressed to larger sizes, intratumoral macrophages were found to increase the expression of iRs, while tumor cells in the same TME also increased their counter-receptors (such as CD47) and the T cell inhibitory molecule PD-L1 ( Figure 6A). These changes in late-stage tumors suggest that they have established stronger immunosuppression compared to their early stages. In the absence of therapy, intratumoral macrophages are constantly connected to surrounding tumor cells through TME immunosuppressive signals, resulting in their iR phosphorylation and binding to SHP-2 rather than SHP-1 in the cytoplasmic ITIM (data not shown). Our cumulative data indicate that this type of iR-SHP-2 binding detains SHP-2 from access to immunosuppressive cytokine receptors (such as IL-4R and IL-10R) and thus prevents SHP-2 from inhibiting anti-inflammatory signal transduction (14). Therefore, this iR-SHP-2 binding in stable solid tumors acts as a feed-forward regulation that enhances the anti-inflammatory phenotype of macrophages, thereby promoting TME immunosuppression and tumor progression.

[0381] SHP-1 activity is low in non-therapeutic solid tumors (see 7A to 7D ), and this low activity inhibits immunosuppressive receptor signaling. In fact, this function of SHP-1 cooperates with SHP-2, both of which are able to bind to IL-4R and IL-10R and inactivate their signaling (see our previous studies and studies by others (14-17). Consistently, inhibition of SHP-1 in MC38 solid tumors leads to increased IL-10 production in the TME ( Figure 6B Interestingly, inhibition of SHP-1 also increased tumor production of IL-6, a cytokine reported to play an immunosuppressive role in the TME and support tumor progression (18,19). In vitro assays of macrophage activation in a tumor-associated environment with the alternative activation (M2) stimuli IL-4, IL-13, and IL-10 confirmed that inhibition of SHP-1 with TPI-1 or another inhibitor, PTP-1, dose-dependently increased macrophage production of IL-10 and TGFβ ( Figure 6C In parallel experiments, the same SHP-1 inhibitor potentiated macrophage responses to the M1 stimuli IFNγ and LPS and enhanced proinflammatory cytokine production.

[0382] Supporting this notion, we found that upon therapeutic treatment of tumors, such as those treated with TLR ligands (αTLR), inflammatory cytokines (IL-1 / 6 / TNFα / IFNγ), STING activators (2'3'-cGAMP), RT, anti-PD-L1 immune checkpoint blockade (αPD-L1), or the chemotherapy drug azacitidine (AZA), a spike in SHP-1 activity was induced ( Figure 7A and Figure 7BIndeed, large, advanced tumors with high iR expression in macrophages provide robust SHP-1 activation under therapeutic treatment. We found that intratumoral macrophages are the primary source of SHP-1 activity, and that depletion of macrophages in tumors following therapy significantly reduced the SHP-1 activity spike. Figure 7C In addition to inactivating pro-inflammatory signaling pathways, these high SHP-1 activities also enhance the production of IL-10 and TGFβ induced by pro-inflammatory stimuli through currently unidentified mechanisms ( Figure 7D , see also Figure 2D ), thus accelerating the restoration of immunosuppression in the TME. Thus, solid tumors with high induction of SHP-1 activity under treatment with TLR agonists or other therapies failed to induce a change in the phenotype of intratumoral macrophages from immunosuppressive to proinflammatory, nor did they reprogram the TME towards antitumor immunogenicity, such as the reprogramming we observed in pancreatic ductal adenocarcinoma (KPC) and colorectal cancer (MC38) under treatment ( Figures 7D to 7F Instead, these tumors become resistant to therapy through increased expression of TGFβ and its receptors (TGFBR1 and TGFBR2), and the chemokine CCL2, which attracts MDSCs ( Figure 7F ), which leads to wound healing and enhanced immunosuppression. In cases of strong treatment resistance, no antigen presentation or only marginal levels of antigen presentation are detected in the TME, and anti-tumor T cell immunity is suppressed.

[0383] In sharp contrast, inhibition of SHP-1 in the presence of a single dose of TPI-1 completely altered how the TME responded to TLR agonists or RT. Just a few hours (6-18h) after treatment (TPI-1+αTLR or RT), intratumoral macrophages changed their phenotype from immunosuppressive to characteristically proinflammatory, characterized by high expression of proinflammatory cytokines such as TNFα, IFNα / β, IFNγ, IL-1β, IL-6, IL-12, IL-17, IL-18, while also reducing IL-10 and TGFβ. A significant immunogenic antigen presentation mechanism was induced, with increases in cell surface markers MHC-I, MHC-II, and co-stimulatory molecules CD80, CD86, CD40, OX40L, etc. A set of chemokines that attract neutrophils, NK, and T cells in the TME was also increased, while TGFR and CCL2 were decreased ( Figures 7D to 7F ).

[0384] Example 6

[0385] Consistent with these changes, TME analysis of MC38 colorectal and KPC pancreatic tumors treated with TLR agonists in combination with TPI-1 or RT ( Figures 8A to 8E9) found that inflammatory neutrophils (ROShigh) and tumor-killing NK cells (Granzhigh) rapid infiltration, and strong antigen presentation also caused the tumor-specific (p15E-reactivity) cytotoxic T cells (Granzhigh) with high tumor destruction ability to expand. The immunosuppressive compartment (including MDSC and Treg) in the same TME is reduced. Interestingly, the intratumoral macrophage (F4 / 80+) population is also reduced to a microsize after T cell activation. We further found that this T cell activation and amplification event is mainly driven by antigen presentation mediated by APC in the tumor of the original site of the tumor-specific memory T cells (TEM / CM) in the TIL. In vitro infusion of TLR agonists and TPI-1 together rather than infusion of each into the resected tumor induces a similar T cell amplification reaction. However, processing the resected tumor does not increase neutrophils and NK cells, thereby indicating that the increase in the infiltration of these killer cells in the tumor in vivo is through chemotaxis recruitment outside the tumor. Together, these results suggest that inhibiting SHP-1 to abrogate a central mechanism across pro-inflammatory signaling therapeutically unleashes intratumoral anti-tumor potential, empowering both innate and adaptive immune cells against cancer.

[0386] In addition, these analyses of changes in the TME immune landscape after TPI-1 combination therapy inform the design of our pulse-intermittent SHP-1 inhibition strategy for metastatic solid tumors. We found in tumors treated with TPI-1 in combination with TLR agonists or RT that the intratumoral macrophage population in the TME was greatly reduced after antigen presentation to activate T cells. In addition to the reduction in macrophages, tumor-associated SHP-1 activity also decreased, a change that eliminates the necessity of continuous TPI-1 application. This "interval" period lasts 3-6 days, and the length depends on the TPI-1 and TLR agonist / RT doses and the tumor type and stage before treatment. During this period, the TME is dominated by tumor-killing CD8 T cells, neutrophils, and NK cells, and the treated tumor growth is limited but presents a state of stable disease (SD) or tumor regression. However, assuming that the tumor is not completely eliminated, the TME is later refilled with "new" macrophages and an increase in MDSC and Treg is shown, thereby indicating that immunosuppression is re-established. Consistent with these changes, tumors resumed growth after the rest period unless another cycle of TPI-1 combination therapy was administered, which again effectively suppressed the tumors.

[0387] Example 7

[0388] Our design of pulsed-intermittent inhibition of SHP-1 (iSHP-1) as an immunoadjuvant / neoadjuvant therapy for solid tumors originated from the above mechanistic studies. Based on our findings, iSHP-1 monotherapy does not provide a rationale, but iSHP-1 combined with a pro-inflammatory regimen predicts potent tumor suppression. Given that intratumoral macrophages indicate TME reactions and SHP-1 activity in solid tumors, the iSHP-1 strategy was adapted to target intratumoral macrophages and exploit their ability to drive pro-inflammatory responses and antigen presentation for tumor elimination. Following the dynamics of the intratumoral macrophage population, iSHP-1 treatment was given in a "pulse" manner when macrophages were abundant in the TME, but was stopped ("interval" period) when they decreased after antigen presentation to activate T cells. In addition, given our finding that macrophages repopulated the TME after the interval period, multiple treatment cycles were designed, assuming that the tumor was not eliminated, with iSHP-1 pulses repeated immediately after the interval period for each cycle.

[0389] FIG. 10A to FIG. 10B Describe the pulse-intermittent iSHP-1 design and treatment regimens. In preclinical tumor models, we have tested three pulse treatment regimens: Pulse-1, -2 and -3, in which iSHP-1 was given continuously once, twice or three times (1x per day) at the beginning of each cycle, and iSHP-1 treatment was stopped during an interval of 2-9 days between each cycle. Combination modalities include but are not limited to TLR ligands, STING activators, RT, anti-PD-1 / L1 immune checkpoint blockers (αPD-1 / L1), inflammatory cytokines, chemotherapy and oncolytic viruses. These combination regimens were given together with SHP-1 inhibitors or after separate dosing schedules. Murine solid tumor models with single tumors or multiple lesions (metastases) in syngeneic mice of different backgrounds were tested. These models were established by implanting tumors in multiple locations and when the size of a single tumor reached ≥200mm 3 or total tumor burden ≥300 mm 3 iSHP-1 treatment was initiated at 4 hr. These models included pancreatic adenocarcinoma (KPC and Pan02), colorectal cancer (MC38), metastatic breast cancer 4T1, lung cancer (LLC), and T-cell lymphoma (EL4). All treatments were administered systemically to mice, either intraperitoneally (ip) or subcutaneously (sc), to achieve a systemic effect on "metastatic" lesions. In a subset of experiments, treatment was also administered via intratumoral injection (it).

[0390] Antitumor efficacy (tumor volume change and survival rate), side effects (weight loss, proteinuria, anemia, and clinical discomfort), macroorgan toxicity and tissue inflammation due to toxicity, and changes in the immune landscape of the TME were analyzed at different time points throughout the treatment and at the final point.

[0391] As seen in the next few examples, we have intensively examined the pulse-intermittent iSHP-1 strategy in preclinical tumor models, and in all cases, pulsing iSHP-1 in combination with a pro-inflammatory regimen induced potent anti-tumor responses and activation of innate and adaptive immune cells to eliminate tumors. Applying this strategy systemically resulted in control and regression of cancer lesions throughout the body, resulting in higher survival rates and durable anti-tumor immunity.

[0392] More importantly, the pulse-intermittent iSHP-1 strategy provides, for the first time, a practical approach that allows SHP-1 inhibition to potentially be implemented as an in vivo treatment regimen, as this strategy minimizes the toxicity caused by SHP-1 deficiency. Compared to animals that were SHP-1 deficient or continuously treated with SHP-1 inhibitors, which developed significant lung inflammation, kidney damage, colitis, anemia, and splenomegaly, mice treated with three cycles of pulse-intermittent iSHP-1, along with TPI-1 at doses sufficient to induce systemic tumor elimination in combination with TLR ligands, STING activators, RT, αPD-1 / L1, and proinflammatory cytokines, did not exhibit severe lung or kidney damage, anemia, or splenomegaly (data are shown in the following section, Case Studies). This high benefit-risk profile is attributed to the pulsed, intermittent iShp-1 design, which is specifically tuned to target intratumoral macrophages for tumor-lesion efficacy and avoids prolonged depletion of SHP-1 activity in essential organs (such as the lungs, intestines, kidneys, and spleen), where SHP-1 activity is critical for preventing unwanted autoimmune inflammatory responses to commensal microorganisms or debris particles. Our further studies support this concept, demonstrating that after pulsed inhibition of SHP-1 in macrophages with the covalent inhibitor TPI-1, macrophages can restore SHP-1 activity within 24 hours.

[0393] Example 8

[0394] We have tested iSHP-1 in combination with TLR agonists (αTLR), STING activators, immune checkpoint blockers (αPD-1 / L1), and / or RT to treat a variety of solid tumors. These include pancreatic ductal adenocarcinoma KPC (KPC-luc) and Pan02, colorectal carcinoma MC38, lung cancer LLC, and metastatic breast cancer 4T1 in immunocompetent murine syngeneic models. Both treatment efficacy and safety were evaluated according to Figure 10.

[0395] Murine Models (single or multifocal solid tumors): 1) pancreatic ductal adenocarcinoma (KPC or KPC-luc)-C57BL6 syngeneic implants, 2) colorectal carcinoma (MC38)-C57BL6 syngeneic implants, 3) lung cancer (LLC or LLC-luc)-C57BL6 syngeneic implants, and 4) metastatic breast cancer (4T1 or 4T1-luc)-BalbC background syngeneic implants.

[0396] Establish tumor model: Healthy cultured cancer cells (1-5x 10 5 ) are injected subcutaneously (sc), intraperitoneally (ip), intravenously (iv) or orthotopically into WT C57BL6 or BalbC mice (6-8 weeks, male and female) to establish a syngeneic model. For multiple tumor lesions (metastasis model), tumor cells are implanted in multiple locations via a combination of multi-point sc injection and ip or iv. After 10-14 days, palpable subcutaneous or orthotopic (e.g., 4T1) tumors are formed. Tumor length and width are recorded using a caliper, and then tumor volume (V) is calculated using the formula: volume = (length x width 2) / 2. For tumor cells expressing luciferase (KPC-luc, LLC-luc, 4T1-luc), whole-body images of luminescence intensity are obtained for presentation of the tumor.

[0397] Therapeutic treatment: When a single tumor grows to ≥200 mm 3 or total tumor burden ≥300 mm 3 Treatment was initiated at 4 hr. Given its potent inhibitory capacity and relative specificity, the covalent SHP-1 inhibitor TPI-1 was selected. For systemic effects at sites distal to the tumor site, varying doses of TPI-1 were administered in PBS via ip or sc injection according to different dosing strategies. In a subset of experiments, the effects of TPI-1 were also tested by direct injection into the tumor (intratumoral injection, i.t.). TPI-1 was administered prior to or in conjunction with the combined modality.

[0398] A. Research-1( Figures 11A to 11E ):Continuous vs. intermittent iSHP-1 and TLR agonist combinations for treatment efficacy and adverse toxicity

[0399] Tumor model: Single implant of KPC pancreatic adenocarcinoma

[0400] Treatment and dosing strategy: i) iSHP-1-TPI-1, 1, 3, and 10 mg / kg; ip, 1x daily (continuous), or intermittently according to the schedule depicted in Figure 11. ii) TLR agonists (αTLR)-CpG, PolyI:C, 10 μg each; ip every 3 days

[0401] Results: TPI-1 exhibited a dose-dependent effect, and its combination with TLR agonists limited KPC pancreatic tumor growth and induced tumor regression. Both continuous and intermittent strategies of TPI-1 administration achieved similar antitumor efficacy. However, continuous administration of TPI-1 (1x daily) caused acute anemia, proteinuria, splenomegaly, and lung inflammation. In mice treated intermittently with TPI-1, these side effects were not apparent or were mild. In summary, the intermittent iSHP-1 strategy greatly reduced the risk of adverse toxicities while achieving tumor inhibitory efficacy.

[0402] B. Study-2( 12A to 12D ): Pulse-intermittent iSHP-1 in combination with TLR agonists and / or ICB (αPD-L1) in multifocal colorectal cancer: a study of efficacy and adverse toxicity

[0403] Conclusions: i) Intermittent iSHP-1 therapy in combination with a TLR agonist (αTLR), or αTLR and αPD-L1, effectively regressed multiple lesions in MC38 colorectal cancer cells that were resistant to either TLR or αPD-L1 alone. ii) TPI-1 administered via ip and sc had similar systemic efficacy. iii) αPD-L1 enhanced iSHP-1 plus αTLR-mediated T cell immune activation. iv) Intermittent iSHP-1 therapy did not produce acute adverse toxicities.

[0404] C. Study-3( 13A to 13E ):Pulse-intermittent iSHP-1 combined with RT and αPD-L1 in the treatment of multifocal pancreatic ductal adenocarcinoma and lung cancer

[0405] Modality and Dosing Strategy: i) iSHP-1-TPI-1, 3 or 5 mg / kg, ip, once every 3 days. ii) Tumor-directed RT: 8 Gy-4 Gy-2 Gy, administered concurrently with iSHP-1 into the right flank. iii) αPD-L1: 100 μg, ip, administered one day after iSHP-1 plus RT.

[0406] Results: Systemic intermittent iSHP-1 combined with right flank tumor-lesion RT, followed by αPD-L1 for T cell-boosting immunity, induced potent antitumor immunity with abscopal effects, effectively eliminating or suppressing KPC pancreatic cancer and LLC lung cancer as well as distant lesions.

[0407] D. Research-4( 14A to 14E ): Pulse-intermittent iSHP-1 combined with TLR agonists treats advanced, large KPC pancreatic cancer.

[0408] Murine Model: Single Large Pancreatic Duct Adenocarcinoma (KPC-luc) C57BL6 Syngeneic

[0409] Treatment: SHP-1 inhibition (iSHP-1) combined with TLR agonists (αTLR)

[0410] Results: As 14A to 14E As shown in , TPI-1 combined with αTLR enhanced anti-tumor immune cells (such as CD8+ T cells, NK cells, and neutrophils) and effectively eliminated advanced, large KPC pancreatic cancer.

[0411] Example 9

[0412] In vitro macrophages were assayed in the presence of tumor cells linked to macrophage iRs with or without TPI-1 inhibition of SHP-1 (iShp1). Figure 16A The test system in Figure 16B and Figure 16C As shown in the results, IFNα treatment alone, without or with iShp1, did not induce antigen presentation or a proinflammatory response in macrophages. IFNγ plus iShp1 significantly increased antigen presentation, even though the combination did not induce proinflammatory cytokine production. In contrast, IL-1 family cytokines (IL-1β, IL-18), TNFα, and TLR ligands, combined with iShp1, exhibited the ability to proinflammatoryally activate macrophages in the tumor environment.

[0413] Example 10

[0414] MC38 colorectal carcinoma was established (via subcutaneous administration) in syngeneic C57BL6 mice. 3 Subsequently, systemic inflammatory disease was induced in tumor-bearing mice by subcutaneous administration of TLR agonists (αTLR, CpG / PolyIC / R848, 25 μg each) at a site distant from the MC38 tumor. TPI-1 (1 mg / kg) was concurrently administered to one group of mice via subcutaneous administration to achieve systemic SHP-1 inhibition.

[0415] like Figure 17A As shown in , SHP-1 protects MC38 tumors from acute inflammation induced by αTLR. Neutrophil infiltration, indicative of local tissue inflammation, was measured in different organs at multiple time points after αTLR stimulation. Inhibition of SHP-1 by TPI-1 unlocks tumor immunosuppression and enables neutrophil infiltration after αTLR (αTLR+TPI-1).

[0416] like Figure 17B As shown in , TEM analysis confirmed increased neutrophil infiltration in tumor tissues in mice treated with αTLR plus TPI-1.

[0417] like Figure 17C As shown in

[15] , inhibition of SHP-1 by TPI-1 enables intratumoral macrophages to skew toward pro-inflammatory activation through αTLRs, resulting in increased expression of TNFα and IL12. Conversely, the absence of TPI-1 causes macrophages to resist pro-inflammatory activation through αTLRs, while enhancing immunosuppression and increasing IL-10 and TGFβ expression.

[0418] Example 11

[0419] This example demonstrates that neutralization of TNFα suppresses systemic inflammation without affecting the anti-tumor efficacy of the combination of TKi, SHP-1 inhibition, and αTLR.

[0420] Mice with established MC38 colorectal cancer (200-400 mm3) were treated with αTLR, TPI-1, and dasatinib (sc) with or without additional treatment with anti-TNFα mAb or anti-IL-6 mAb (150 μg, ip). Treatment was repeated once (d1 and d2). Tumor volume changes were recorded, and immune infiltration of the tumor TME was analyzed on day 6 after treatment. Figure 11A .

[0421] like Figure 11B As shown in , tumor volume decreased after αTLR+TPI-1+dasatinib treatment, and administration of anti-TNFα or anti-IL-6 did not interfere with its antitumor activity. Treatment with anti-TNFα mAb or anti-IL-6 mAb also did not affect the increase in CD8 T cells (Tc) and NK cells and the decrease in macrophages and MDSCs induced by αTLR / TPI-1 / dasatinib therapy in the TME. Figure 11C and Figure 11D Treatment of mice with anti-TNFα mAb, but not anti-IL-6 mAb, largely reduced the induction of inflammatory cytokines (TNFα, IL-6, IL-1β, IL-10, IFNα, and IFNγ) associated with the αTLR / TPI-1 / dasatinib combination therapy. Anti-TNFα treatment also significantly reduced circulating monocyte and PMN chemokines CCL2, CCL5, and CXCL1, while not reducing CXCL10, which is essential for T cell trafficking. Figure 11E Furthermore, as shown in FIG11F , anti-TNFα treatment protected mice from developing splenomegaly and intestinal inflammation typically associated with αTLR / TPI-1 / dasatinib therapy.

[0422] In summary, the results show that neither anti-TNFα nor anti-IL-6 interferes with TKi / iShp1 / αTLR-driven anti-tumor immunity or therapeutic efficacy. In addition, anti-TNFα exhibits beneficial effects by largely abrogating the TKi / iShp1 / αTLR-induced cytokine storm and systemic inflammation, thereby suppressing the adverse toxicities associated with therapy.

[0423] Furthermore, although the specific experiments above involved the use of both TPI-1 and dasatinib and both polyI:C and R848, our results from experiments using either TPI-1 or dasatinib or either polyI:C or R848 achieved similar effects (data not shown). We also found that the appropriate time window for anti-TNFα antibody treatment can be at least one week prior to SHP-1 inhibitor / αTLR treatment (as long as the antibody is stable during the time window) to immediately after (e.g., within 0.5-1 hour). Preferably, the anti-TNFα antibody is provided prior to or concurrently with the SHP-1 inhibitor and / or αTLR so that it maximally blocks TNFα induced after treatment with the SHP-1 inhibitor and proinflammatory agent.

[0424] Example 12

[0425] The mechanism by which tumor cells inhibit the pro-inflammatory response of macrophages in the TME was investigated. As shown in Figure 19A, macrophage responses in the tumor environment were measured: human monocyte-derived macrophages were injected into the tumor Cells were cultured alone (configuration 1), in the presence of cancer cell-conditioned medium (50%) containing cancer cell-secreted factors (secretome; configuration 2), or co-cultured with cancer cells (cell “contact” model; configuration 3), or in the absence of “contact” Macrophages in these environments were treated with the pro-inflammatory stimuli TLR agonist R848 (1 μg / ml), R848 plus IFNγ (40 ng / ml), or the sting activator MSA-2 (10 μg / ml).

[0426] We then examined macrophage responses to R848 / IFNγ. The presence of the cancer secretome (profiles 2 and 4) did not alter the proinflammatory response of macrophages, even though it resulted in a partial suppression of proinflammatory cytokine production (10%-40% reduction). In contrast, macrophages co-cultured with cancer cells exhibited enhanced immunosuppression and high IL-10 production, despite a reduction in proinflammatory cytokine production (Figure 19, Panel B).

[0427] Macrophage responses to R848 or MSA-2 were also investigated. Results were similar to those in Panel B: the presence of the cancer secretome partially suppressed the macrophage proinflammatory response, but macrophages co-cultured with cancer cells exhibited abrogated proinflammatory responses and enhanced immunosuppression with elevated IL-10 and TGFβ production. See Figure 19, Panel C.

[0428] In addition, if Figure 20 As shown in , macrophages co-cultured with cancer cells abrogated macrophage antigen presentation induced by a TLR agonist (R848) plus IFNγ.

[0429] Example 13

[0430] It has been found that when the tumor progresses to the late stage (large size), iR and its ligands are upregulated, as shown in Figure 21. Panel A depicts the composition and percentage of immune cells within a typical MC38 colorectal cancer. Panel B depicts the expression of multiple inhibitory receptors (iR) on the myeloid immune population (including TAM (F4 / 80+), MDSC (Ly6C+) and N2- neutrophils (PMN)) in MC38 carcinomas of different sizes. Panels C to D depict the expression of CD47 (a receptor for SIRPα) and PD-L1 on the same MC38 carcinomas of different sizes as in Panel B. Panel E depicts representative IHC staining of samples of human cancers in which the expression of ligands for myeloid iRs was increased.

[0431] Example 14

[0432] It was also found that factors (secreted protein groups) produced by cancer cells and tumor TME induce increased macrophage expression of iR. See, for example, Figure 22. Figure A shows increased expression of iR on bone marrow leukocytes in murine solid tumors (including 4T1 breast cancer, LLC lung cancer, and EL4 T cell lymphoma). Treatment of murine bone marrow-derived macrophages with murine cancer cell-conditioned medium induces increased expression of iR. Examples of cancer cells are B16 melanoma cells, MC38 colorectal cancer cells, EL4 T cell lymphoma cells, and LLC lung cancer cells. SIRPα is an example of iR. See Figure B of Figure 22.

[0433] Treatment of human monocyte-derived macrophages with human cancer cell conditioned medium induces increased iR expression. Examples of cancer cells shown are: colorectal cancer cells T84, HT29 and SW620T, breast cancer cells T47D, lung cancer A549, renal cancer TK10, ovarian cancer OVCAR3 and monocytic leukemia THP1. Examples of iR tested on human macrophages are: SIRPα, LILRB1, LILRB4 and Siglec7. See Figure C of Figure 22. Cytokines produced by murine and human cancer cells in culture are shown in Figure D of Figure 22. Treatment of macrophages with cancer cell cytokines induces increased iR expression on macrophages (bone marrow-derived macrophages (BMDM) and peritoneal macrophages (PEM)) (SIRPα as an example). See Figure E of Figure 22.

[0434] Example 15

[0435] Inhibition of SHP-1 unleashes pro-inflammatory responses in the KPC tumor TME, such as FIG. 23A to FIG. 23B As shown in . KPC pancreatic tumors removed from mice were cut into small pieces and treated with 5 different TLR agonists (R848, 3M-852A, motomod, bropirimide, or visammod; each 1 μg / ml) without or with the SHP-1 inhibitor TPI-1 (0.4 μM). After 18 h, cytokine secretion in the culture medium was measured.

[0436] like Figure 23A As shown in

[16] , all five TLR agonists failed to trigger proinflammatory cytokine production but induced high levels of IL-10 and TGFβ, suggesting enhanced immunosuppression in the TME.

[0437] like Figure 23B As shown in (A), inhibition of SHP-1 (iSHP-1) enables TLR agonists to drive proinflammatory responses in tumor tissue. As shown, addition of the SHP-1 inhibitor TPI-1 to (A) induces high proinflammatory cytokine production while inhibiting the production of IL-10 and TGFβ.

[0438] Example 16

[0439] Treatment of MC38 tumors with TLR agonists (αTLR) plus SHP-1 inhibition induces pro-inflammatory polarization of the TME. MC38 colorectal tumors excised from mice were cut into small pieces and treated with CpG+Poly(I:C)+R848 (0.4 ug / ml each) ± TPI-1 (0.4 μM). After 18 h, cytokine secretion in the culture medium was measured. Figure 24 As shown in

[16] , αTLR plus TPI-1 induced high levels of proinflammatory responses while suppressing IL-10 in the TME.

[0440] Example 17

[0441] Study on the iR→SHP-1 inhibitory axis. SHP-1 Dephosphorylates inflammatory stimulus-induced JAK-STAT, NFκB, MAPK, and PI3K-Akt activation pathways, thereby quenching pro-inflammatory signaling and conferring therapeutic resistance. SHP-1 Inhibition of α-glucose-mediated cytotoxicity abolishes multi-axis iR-mediated inhibitory regulation, thereby releasing macrophage pro-inflammatory polarization, such as FIG. 25A to FIG. 25B As shown in .

[0442] Example 18

[0443] like Figure 26B and Figure 26C As shown in Figure 2, under TLR and IFNγ stimulation, Shp1 - / - Macrophages resist the inhibition imposed by cancer cells and release a proinflammatory response. In the presence of B16 melanoma cells, cells expressing WT or isotype-bound Shp1 were treated with a TLR agonist (αTLR; R848, 1 μg / ml) plus IFNγ (40 ng / ml). - / - Mouse bone marrow-derived macrophages (BMDM) are prepared from mice. Figure 26A After 16 h, the cell culture medium was collected and cytokines were determined. The results are shown in Figure 26B Macrophages were also collected and their inflammatory phenotype and cell surface expression of antigen presentation machinery were determined. The results are shown in Figure 26C middle.

[0444] Example 19

[0445] like Figures 27A to 27C As shown in , cell surface blockade of iRs or ligands is an alternative strategy to deplete the iR→SHP-1 axis.

[0446] Experimental setup: Human macrophages co-cultured with cancer cells were stimulated with proinflammatory factors (e.g., TLR agonists, IFNγ, Sting activators, etc.) in the presence or absence of TPI-1 (inhibition of SHP-1, or iSHP-1), or mAbs that block Siglec (αSiglec-7, -8, or -9), mAbs that block LilRB-MHC interactions (αLILRB1, αLilRB2, αLIlRB3, αLilRB4, or pan-HLA-A / B / C blocking Abs), or mAbs that block CD47-SIRPα interactions (αCD47 or αSIRPα). To remove cancer cell surface sialic acid structures that bind to Siglec on macrophages, cancer cells were also treated with neuraminidase (50 mU / ml, 1 h) prior to use in the experiment. See Figure 27A.

[0447] The information of the blocking antibodies used in the experiments is shown in Figure 27B These antibodies are commercially available and used at 2-10 μg / ml.

[0448] An example of data (macrophages and SW260 cancer cells). Figure 27C As shown in , blockade of a single iR-ligand axis is insufficient to remove the immunosuppression imposed by cancer cells on macrophages, whereas blocking multiple iR-ligand interactions or inhibition of SHP-1 downstream of all iRs in macrophages eliminates tumor cell suppression, thereby releasing macrophages for a proinflammatory response. Similar results were obtained when macrophages were co-cultured with other cancer cells (e.g., OVCAR3, MDA231, TK10, HT29, etc.).

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Claims

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

2. A method of treating cancer in an individual, the method comprising administering to the individual a) a SHP-1 inhibitor, and b) a pro-inflammatory agent, wherein the method comprises systemically administering the SHP-1 inhibitor.

3. A method of treating cancer in an individual, the method comprising administering to the individual a) a SHP-1 inhibitor, and b) a proinflammatory agent, wherein the proinflammatory agent comprises an agent selected from the group consisting of a TLR agonist, a STING activator, a PAMP / DAMP activator, chemotherapy, a proinflammatory cytokine, a cancer vaccine, a bacterial component, acoustic wave therapy, magnetic therapy, electrical therapy, and electrostatic therapy.

4. A method of treating cancer in a subject, the method comprising administering to the subject a SHP-1 inhibitor, wherein the subject is experiencing an inflammatory response.

5. The method of any one of claims 2-4, wherein the method further comprises intermittently administering the SHP-1 inhibitor to the individual.

6. The method of any one of claims 1 and 3-5, wherein the method comprises systemically administering the SHP-1 inhibitor.

7. The method of any one of claims 1, 5, and 6, wherein the method comprises administering the SHP-1 inhibitor at least twice at intervals no more than once every three days.

8. The method of any one of claims 1 and 5-7, wherein the method comprises administering the SHP-1 inhibitor to the individual for at least two cycles, wherein each cycle has about three to about twenty days.

9. The method of any one of claims 1-8, wherein the SHP-1 inhibitor does not inhibit SHP-2.

10. The method of any one of claims 1-9, wherein the half-life of the SHP-1 inhibitor is no more than about 5 days, optionally the half-life of the SHP-1 inhibitor is no more than about 3 days.

11. The method of any one of claims 1-10, wherein the SHP-1 inhibitor is effective to inhibit greater than 50% of SHP-1 activity for no more than about 5 days, optionally wherein the SHP-1 inhibitor is effective to inhibit greater than 50% of SHP-1 activity for no more than about 3 days.

12. The method of any one of claims 1-11, wherein the SHP-1 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 SHP-1 or activate SHP-1).

13. The method of claim 12, wherein the SHP-1 inhibitor is selected from the group consisting of TPI-1 or an analog or derivative thereof, a vitamin E derivative, Phomopsis xanthone A (PXA), and a PKC theta activator.

14. The method of claim 13, wherein the SHP-1 inhibitor comprises TPI-1.

15. The method of any one of claims 1-14, wherein the SHP-1 inhibitor is administered at least three times.

16. The method of any one of claims 1-15, wherein the method comprises systemic and local administration of the SHP-1 inhibitor, optionally wherein the method comprises intratumoral administration of the SHP-1 inhibitor.

17. The method of any one of claims 2 and 6-16, wherein the systemic administration of SHP-1 comprises oral administration, intravenous administration, subcutaneous administration, and / or intraperitoneal administration.

18. The method of any one of claims 1-3 and 5-17, wherein the proinflammatory agent and the SHP-1 inhibitor are administered within 24 hours of each other, optionally wherein the proinflammatory agent and the SHP-1 inhibitor are administered within 4 hours of each other.

19. The method of any one of claims 1-3 and 5-18, wherein the method comprises administering the proinflammatory agent intratumorally.

20. The method of any one of claims 1-3 and 5-19, wherein the method comprises administering the proinflammatory agent to a site different from the site of the cancer to be treated.

21. The method of any one of claims 1-2 and 5-18, wherein the proinflammatory 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 proinflammatory cytokine, a chemotherapeutic agent, a bacterial component, a cancer vaccine, an oncolytic virus, acoustic wave therapy, magnetic therapy, electrical therapy, and electrostatic therapy.

22. The method of any one of claims 1-3 and 5-21, wherein the proinflammatory agent comprises a TLR agonist.

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

24. The method of claim 23, wherein the TLR agonist comprises CpG, polyI:C and / or R848.

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

26. The method of any one of claims 1-3 and 5-25, wherein the pro-inflammatory agent comprises a STING activator.

27. The method of claim 26, wherein the STING activator comprises 2'3'-cGAMP.

28. The method of any one of claims 1-3 and 5-27, wherein the pro-inflammatory agent comprises a chemotherapeutic agent.

29. The method of claim 28, wherein the chemotherapy comprises azathioprine (AZA).

30. The method of any one of claims 1-3 and 5-29, wherein the proinflammatory agent comprises a proinflammatory cytokine.

31. The method of claim 30, wherein the proinflammatory cytokines comprise IL-1b, IL-18, IL-6 and / or TNFα.

32. The method of any one of claims 1-2 and 5-31, wherein the proinflammatory agent comprises radiation therapy.

33. The method of claim 32, wherein the radiation therapy comprises irradiating the site of the cancer to be treated.

34. The method of claim 32 or claim 33, wherein the radiation therapy comprises irradiating a site different from the site of the cancer to be treated.

35. The method of any one of claims 32-34, wherein the dose of radiation therapy is insufficient to kill tumor cells.

36. The method of any one of claims 1-2 and 5-35, wherein the proinflammatory agent comprises a checkpoint inhibitor.

37. The method of claim 36, wherein the checkpoint inhibitor comprises an anti-PD-L1 antibody, an anti-PD-1 antibody, or an anti-CLTA4 antibody.

38. The method of any one of claims 1-3 and 5-37, wherein the proinflammatory agent is administered intermittently.

39. The method of any one of claims 1-3 and 5-38, wherein the proinflammatory agent and the SHP-1 inhibitor are administered simultaneously or concurrently.

40. The method of any one of claims 1-3 and 5-39, wherein the pro-inflammatory agent comprises an immune cell.

41. The method of any one of claims 4-39, wherein the method further comprises an immune cell.

42. The method of claim 41, wherein the immune cells are derived from the same individual.

43. The method of claim 41 or claim 42, wherein the immune cell comprises or is a macrophage, optionally wherein the macrophage has an M1 phenotype.

44. The method of any one of claims 40-43, wherein the immune cells are derived from monocytes.

45. The method of any one of claims 40-44, wherein the immune cells express high levels of MHC-I, MHC-II, CD80 and / or CD86.

46. The method of any one of claims 40-45, wherein the immune cells express one or more proinflammatory cytokines, optionally wherein the one or more proinflammatory cytokines comprise TNFα and / or IL-12.

47. The method of any one of claims 40-46, wherein the immune cells do not express significant levels of TGFβ and / or IL-10.

48. The method of any one of claims 40-47, wherein the immune cells comprise T cells.

49. The method of any one of claims 40-48, wherein the immune cell is engineered to express a chimeric antigen receptor, optionally wherein the chimeric antigen receptor specifically binds to a tumor antigen.

50. The method of any one of claims 43-49, wherein the macrophages are engineered to be defective in SHP-1 expression and / or activation.

51. The method of any one of claims 40-50, wherein the SHP-1 inhibitor and the immune cell are administered within 24 hours of each other, optionally wherein the SHP-1 inhibitor and the immune cell are administered within 4 hours of each other.

52. The method of any one of claims 40-51, wherein the immune cells are administered simultaneously or concurrently with the SHP-1 inhibitor.

53. The method of any one of claims 1-52, further comprising administering to the individual an effective amount of an anti-TNFα antibody.

54. The method of any one of claims 1-53, wherein the cancer is a solid tumor.

55. The method of any one of claims 1-53, wherein the cancer is a hematological cancer.

56. The method of any one of claims 1-55, wherein the cancer is advanced cancer.

57. The method of any one of claims 1-56, wherein the cancer is resistant or refractory to radiation therapy, chemotherapeutic agents, and / or checkpoint inhibitors.

58. The method of any one of claims 1-57, wherein the individual is a human.

59. A composition comprising a SHP-1 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 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 acoustic wave therapy, magnetic therapy, electrical therapy, or electrostatic therapy.

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