TW202421172A - Shp-1 inhibitors for treating cancer - Google Patents

Abstract

The present application provides methods of treating a cancer in an individual that involves administering to the individual a SHP-1 inhibitor (such as TPI-1 or an analog or a derivative thereof) and a pro-inflammatory agent (such as a TLR agonist, a radiation therapy or an immune checkpoint inhibitor). In some cases, the methods involves administering a SHP-1 inhibitor when the individual is under an inflammation reaction.

Description

SHP-1 inhibitors for cancer treatment

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.

In cancers such as solid tumors, intratumoral myeloid leukocytes, including macrophages (i.e., tumor-associated macrophages or TAMs) and myeloid-derived suppressive cells (MDSCs), play a key role in controlling tumor microenvironment (TME) immunosuppression that supports tumor growth and also confers tumor resistance to immunotherapeutic treatments. An important mechanism by which intratumoral myeloid leukocytes adapt 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) in their cytoplasmic domain leads to activation of the central signaling regulator SHP-1, which mediates dephosphorylation and thus inactivation of multiple signaling molecules, thereby attenuating the anti-cancer proinflammatory response induced by the therapeutic agent (Figure 1). In solid tumors, essential cell surface iRs (such as SIRPα, Siglec, LilRB, PirB, LAIR1, lectin receptors, SLAM family receptors, etc. (1, 2)) that also show an increase in TME expression as tumors progress to advanced stages are regulated by activated SHP-1, which then mediates downstream inhibition.

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

The disclosures of all publications, patents, patent applications, and published patent applications mentioned herein are hereby incorporated by reference in their entirety.

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 pro-inflammatory 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 pro-inflammatory agent comprises an agent selected from the group consisting of: TLR agonists, STING activators, radiation therapy, PAMP/DAMP molecules, checkpoint inhibitors, pro-inflammatory cytokines, pro-inflammatory cells, cells, cancer vaccines, chemotherapeutic agents, bacterial components, cancer vaccines, oncolytic viruses, sound wave therapy, magnetic therapy, electrical therapy, and electrostatic therapy.

In another aspect, the present application provides a method for 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: TLR agonists, STING activators, radiation therapy, PAMP/DAMP activators, checkpoint inhibitors, proinflammatory cytokines, chemotherapeutics, bacterial components, cancer vaccines, oncolytic viruses, sound wave therapy, magnetic therapy, electrical therapy, and electrostatic therapy.

In another aspect, the present application provides a method for treating cancer in an individual, comprising administering to the individual a) a SHP-1 inhibitor, and b) a pro-inflammatory agent, wherein the pro-inflammatory agent comprises an agent selected from the group consisting of: a TLR agonist, a STING activator, a PAMP/DAMP activator, chemotherapy, a pro-inflammatory cytokine, a cancer vaccine, a bacterial component, sonic 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.

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 in 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 immune cells.

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

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

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.

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 longer than about 5 days, optionally wherein the SHP-1 inhibitor is effective to inhibit greater than 50% of SHP-1 activity for no longer than about 3 days.

In some embodiments according to 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., antibody agents that target SHP-1 or activate SHP-1), or protein agents containing SH2 domains (inhibiting SHP-1 activation by competing with ITIM motifs), 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 its analogs or derivatives, vitamin E derivatives, phomoxanthone dimer A (PXA), and PKCθ activators. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

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.

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

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 (e.g., within about 16 hours, 8 hours, 4 hours, 2 hours, 1 hour, or 0.5 hours).

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

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.

In some embodiments according to any of the methods described above, the proinflammatory agent comprises a TLR agonist. In some embodiments, the TLR agonist activates TLRs on macrophages. In some embodiments, the TLR comprises TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8 and/or TLR9. In some embodiments, the TLR agonist comprises CpG, poly I:C (polyI:C) and/or R848, flagellin (TLR5), yeast polysan (TLR2/4), DAMPs produced by radiation therapy (such as HMGB1 (TLR2/4)), DNA and RNA molecules (TLR3/7/8/9), etc.

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).

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.

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).

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).

In some embodiments according to any of the methods described above for use, 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 insufficient to eliminate the tumor (kill all tumor cells).

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.

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

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

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 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 be defective in 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.

In some embodiments of 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 interleukin cascade or interleukin 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 interleukin cascade or interleukin storm is administered simultaneously with a tyrosine kinase inhibitor. In some embodiments, the agent that reduces systemic inflammation and/or reduces the inflammatory interleukin cascade or interleukin storm is administered sequentially with (e.g., before or after) the tyrosine kinase inhibitor. In some embodiments, the agent that reduces systemic inflammation and/or reduces the inflammatory interleukin cascade or interleukin storm is administered on the same dosing schedule as the tyrosine kinase inhibitor.

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

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

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

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

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

In another aspect, the present application provides a composition comprising a SHP-1 inhibitor and a proinflammatory agent, optionally comprising an agent selected from the group consisting of: immune cells, TLR agonists, STING activators, agents for radiation therapy, PAMP/DAMP activators, checkpoint inhibitors, proinflammatory cytokines, chemotherapeutic agents, bacterial components, cancer vaccines, oncolytic viruses, and agents for ultrasound therapy, magnetic therapy, electrical therapy or electrostatic therapy.

Cross-references to related applications

This application claims the benefit of and priority to U.S. Provisional Application No. 63/404,392 filed on September 7, 2022 and U.S. Provisional Application No. 63/491,008 filed on March 17, 2023, the contents of each of which are hereby incorporated by reference in their entirety.

In one aspect, the present application provides a method for treating cancer in an individual, 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 for treating cancer in an individual, comprising administering to the individual monocytes or macrophages with a defect 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 of no more than once every three days. In some embodiments, the method comprises administering a SHP-1 inhibitor to a subject 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, sonic therapy, magnetic therapy, electrical therapy, and electrostatic therapy.

The present application is based, at least in part, on the surprising discovery that combining the "master" inhibitory executive SHP-1 with pro-inflammatory therapy releases pro-inflammatory signaling in the tumor environment, particularly on tumor-infiltrating macrophages, leading to a powerful 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 therapy because such treatments often induce hyperphosphorylation of ITIMs, thereby promoting "hyperactivation" of SHP-1 (a feedback loop that protects tumors from therapeutic damage and inflammatory insults (ahead)) and also induces wound healing responses to promote tumor progression. See, e.g., Figures 1 and 5A. This finding emphasizes the necessity of inhibiting SHP-1 as a combination in tumor immunotherapy in order to be effective.

It has been demonstrated that combining a SHP-1 inhibitor (e.g., TPI-1) with a pro-inflammatory agent (e.g., TLR agonists, pro-inflammatory interleukins, radiation therapy, checkpoint inhibitors) achieves the amazing effect of transforming an immunosuppressive TME into an inflammatory TME, thereby energizing multiple types of immune cells (e.g., macrophages, T cells, and B cells) and completely depleting the tumor. See, e.g., FIG. 11C and FIG. 12B. It has also been demonstrated that this combination therapy achieves a distal effect (e.g., FIG. 13B) and that it is effective in treating advanced, larger-sized tumors (e.g., FIG. 14C).

In addition, it has been found that when administered in intermittent dosing, SHP-1 inhibitors are able to achieve a significant anti-tumor effect comparable to that of SHP-1 inhibitors administered in continuous dosing, while achieving significantly fewer side effects (such as anemia, kidney damage, and liver damage). See Figures 11A to 11D. This is very surprising in view of the severe side effects associated with SHP-1 inhibitors demonstrated in previous studies. In addition, administration of agents that reduce systemic inflammation (e.g., anti-TNFα mAbs) further inhibits systemic inflammation and reduces adverse toxicity. See Figures 18A to 18G.

Therefore, the present application provides a novel approach that can effectively rewire the immunosuppression imposed by tumor pathology and allow innate and adaptive immunity against cancer, thereby achieving significant anti-tumor efficacy. I. Definitions

In general, the terms used in the claims and this specification are intended to be interpreted as having the ordinary meanings as understood by those 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.

The terms "individual," "subject," or "patient" are used synonymously herein to describe mammals, including humans. Individuals include, but are not limited to, humans, cattle, horses, cats, dogs, rodents, or primates. In some embodiments, the individual is a human. In some embodiments, the individual suffers from a disease, such as cancer. In some embodiments, the individual is in need of treatment.

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.

As used herein, the term "intermittent" or "intermittently" in the context of dosing refers to non-continuous dosing, such as shown in Figure 11A (bottom), Figure 12A, Figure 13A, and Figure 14A. In some cases, "intermittent" dosing refers to dosing in which a) the SHP-1 inhibitor is administered for less than 12 consecutive days (e.g., less than 11, 10, 9, 8, 7, 6, 5, 4, and 3 days), and b) the SHP-1 inhibitor is administered at least twice, and the two administrations are 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 per day for no more than three consecutive days and separated by at least one day.

As used herein, the term "cycle" in the context of dosing refers to a period of time during which there is at least one administration of a SHP-1 inhibitor. Day 1 of a cycle is defined as the day when the first administration of a SHP-1 inhibitor occurs during that period. When there are several daily continuous administrations of a SHP-1 inhibitor, Day 1 of a cycle is defined as the day when the first administration occurs among the several daily continuous administrations. The last day of a cycle is defined as the day before the next non-continuous administration of a SHP-1 inhibitor occurs. See Figures 12A and 14A for exemplary cycles. Cycles do not have to 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, the first cycle (which may have five days) may have one SHP-1 inhibitor administration, and the second cycle (which may have seven days) may have two SHP-1 inhibitor administrations.

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

As used herein, "treatment" or "treating" is an approach for obtaining beneficial or desired results (including clinical results). For the purposes of the present invention, beneficial or desired clinical results include, but are not limited to, one or more of the following: alleviation of one or more symptoms caused by a disease, reduction in the severity of a disease, stabilization of a disease (e.g., preventing or delaying worsening of a disease), prevention or delay of the spread of a disease (e.g., metastasis), prevention or delay of the onset or recurrence of a disease, delay or slowing the progression of a disease, improvement of disease symptoms, remission of a disease (whether partial or total), reduction in the dosage of one or more other drugs required to treat a disease, delaying the progression of a disease, improving quality of life and/or prolonging survival. "Treatment" also encompasses the alleviation of the pathological consequences of cancer. The methods of the present invention encompass any one or more of these treatment aspects.

As used herein, "delaying" the development of cancer means slowing, impeding, slowing, arresting, stabilizing and/or postponing the development of the disease. This delay can be of varying lengths of time, depending on the medical history and/or the individual being treated. As will be apparent to one skilled in the art, a sufficient or significant delay may actually encompass prevention, such that the individual does not develop the disease. A method that "delays" the development of cancer is one that reduces the chance of disease development within a given time frame and/or reduces the severity of the disease within a given time frame, 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 progression can be detected using standard methods, including but not limited to computed tomography (CAT scan), magnetic resonance imaging (MRI), abdominal ultrasound, coagulation testing, arteriovenous imaging, or biopsy. Progression can also refer to the progression of cancer that may not be initially detectable and includes onset, recurrence, and relapse.

As used herein, the term "simultaneous administration" means that the first therapy and the second therapy 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).

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 20 minutes, 30 minutes, 40 minutes, 50 minutes, 60 minutes or more). The first or second therapy may be administered first. The first and second therapies are contained in separate compositions, which may be contained in the same or different packages or kits.

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

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 for administration to a subject without causing any significant undesirable biological effect or interacting in a deleterious manner with any other component of the composition in which it is contained. Pharmaceutically acceptable carriers or formulations preferably meet the required standards for toxicology and manufacturing testing and/or are included in the Inactive Ingredient Guide established by the U.S. Food and Drug Administration.

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

As used herein, reference to "about" a value or parameter includes (and describes) variations with respect to that value or parameter itself. For example, a description referring to "about X" includes a description of "X".

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

As used herein, the term "about X-Y" has the same meaning as "about X to about Y".

It should 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.

Any term not directly defined herein shall be understood to have a meaning generally associated with that understood in the art of the present invention. Certain terms are discussed herein to provide additional guidance to practitioners in describing the compositions, devices, methods, and the like of 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 terms and synonyms may be used for any one or more of the terms discussed herein. It will not be important whether a term is detailed or discussed herein. Some synonyms are provided or methods, materials, and the like may be substituted. Unless expressly stated, the recitation 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 herein. II. Treatment methods

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

In some embodiments, the method comprises 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, as appropriate, 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 systemic 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 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 a) has received, is receiving, or will receive a pro-inflammatory agent (e.g., a TLR agonist, e.g., R848, e.g., radiation therapy), or b) is in an inflammatory response or has an ongoing 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, as appropriate, 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 SHP-1 inhibitor is administered at intervals of no more than once every two days. In some embodiments, the SHP-1 inhibitor is administered no less than twice and no more than 5 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 a pro-inflammatory 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 2 weeks (e.g., within 10 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or the same day). In some embodiments, the 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 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., antibody agents that target 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, pseudopteroylanthrone dimer A (PXA), and a PKCθ activator. In some embodiments, the method further comprises administering a pro-inflammatory agent locally (e.g., intratumorally) to the subject. In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and/or reduces the inflammatory interleukin cascade or interleukin 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 pro-inflammatory agent comprises an agent selected from the group consisting of R848, 3M-852A, Motolimod, Bropirimine, and Vesatolimod. In some embodiments, the pro-inflammatory agent comprises a TLR agonist (e.g., R848) and a pro-inflammatory cytokine (e.g., IFN-γ). In some embodiments, the SHP-1 inhibitor comprises TPI-1.

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 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 the SHP-1 inhibitor to the individual at least twice at intervals no more than once every three days. In some embodiments, the SHP-1 inhibitor is administered twice (e.g., two implementation days) every seven to twenty days. In some embodiments, the SHP-1 inhibitor is administered three times (e.g., three implementation days) every ten to twenty days. In some embodiments, the SHP-1 inhibitor is administered at intervals no more than once every two days. In some embodiments, the SHP-1 inhibitor is administered no less than twice and no more than 5 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 2 weeks (e.g., within 10 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or the same day). In some embodiments, the half-life of the SHP-1 inhibitor does not exceed about 10 days (e.g., does not exceed 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 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, pseudopteroylanthrone dimer A (PXA), and a PKCθ activator. In some embodiments, the method further comprises administering a pro-inflammatory agent locally (e.g., intratumorally) to the subject. In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and/or reduces the inflammatory interleukin cascade or interleukin 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 pro-inflammatory agent comprises an agent selected from the group consisting of 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.

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 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 the SHP-1 inhibitor to the 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 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 2 weeks (e.g., within 10 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or the same day). In some embodiments, the 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, pseudo-piperidin dimer A (PXA), and a PKCθ activator. In some embodiments, the method further comprises administering a pro-inflammatory agent locally (e.g., intratumorally) to the subject. In some embodiments, the method further comprises administering to the subject an agent that reduces systemic inflammation and/or reduces the inflammatory interleukin cascade or interleukin 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 an agent selected from the group consisting of 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.

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 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) intravenously, subcutaneously, and/or intratumorally to the subject, optionally wherein the SHP-1 inhibitor is effective to inhibit SHP-1 activity by more 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 of 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, as appropriate, 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 SHP-1 inhibitor is administered at intervals of no more than two times every seven to twenty days. In some embodiments, the SHP-1 inhibitor is administered at intervals of no more than three times every seven to twenty days. In some embodiments, the SHP-1 inhibitor is administered at intervals of 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 2, 3, 4, 5, or 6 times within 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 2 weeks (e.g., within 10 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or the same day). In some embodiments, the 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 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., 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, pseudopteroylxanthrone dimer A (PXA), and a PKCθ activator. In some embodiments, the method further comprises administering a proinflammatory agent locally (e.g., intratumorally) to the subject. 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 subject an agent that reduces systemic inflammation and/or reduces the inflammatory interleukin cascade or interleukin 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 an agent selected from the group consisting of 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.

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 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) intravenously, subcutaneously, and/or intratumorally to the subject, wherein the SHP-1 inhibitor is effective to inhibit SHP-1 activity by more 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, as appropriate, 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 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 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 2 weeks (e.g., within 10 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or the same day). In some embodiments, the 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, pseudopteroylxanthrone dimer A (PXA), and a PKCθ activator. In some embodiments, the method further comprises administering a proinflammatory agent locally (e.g., intratumorally) to the subject. 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 subject an agent that reduces systemic inflammation and/or reduces the inflammatory interleukin cascade or interleukin 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 an agent selected from the group consisting of 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.

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 (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 immune cell described herein) to the subject. In some embodiments, the subject 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 subject is in an inflammatory response or has an ongoing 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 to the individual (e.g., intravenously, subcutaneously, and/or intratumorally) a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof), a proinflammatory agent (e.g., a TLR agonist, e.g., R848, e.g., radiation therapy), and immune cells. 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., neutrophils expressing CAR). In some embodiments, the immune cells comprise antigen presenting cells (APCs). In some embodiments, the immune cells are engineered to express chimeric receptors that specifically bind to tumor antigens. 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 at least twice (separated by at least one day), as appropriate. 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 a SHP-1 inhibitor is separated from the previous or subsequent administration of a SHP-1 inhibitor by at least one day. In some embodiments, a SHP-1 inhibitor, immune cells, and/or a proinflammatory agent are administered within 7, 6, 5, 4, 3, 2, or 1 day. In some embodiments, a 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, a SHP-1 inhibitor, immune cells, and/or a proinflammatory agent are administered simultaneously. In some embodiments, a SHP-1 inhibitor, immune cells, and/or a proinflammatory agent are administered in parallel. In some embodiments, the SHP-1 inhibitor, immune cells and/or pro-inflammatory agents are administered sequentially. In some embodiments, the SHP-1 inhibitor is administered systemically and the pro-inflammatory 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 interleukin cascade or interleukin 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 an agent selected from the group consisting of 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.

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, as appropriate, 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 next administration of the SHP-1 inhibitor by at least one day. 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 hour, or within 30 minutes). In some embodiments, the method comprises administering to the individual a SHP-1 inhibitor 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 a subject for at least two cycles, wherein the SHP-1 inhibitor is administered at least once (e.g., at least twice 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 one of TLR2, TLR3, TLR4, TLR5 and TLR6, optionally wherein the TLR agonist comprises diacylated lipopeptide, heat shock protein, HMGB1, uric acid, fiber binding protein and/or ECM protein. In some embodiments, the TLR agonist activates TLR2, optionally wherein the TLR agonist comprises Pam3Cys, SMP-105 and/or CBLB612. In some embodiments, the TLR agonist activates TLR3, optionally wherein the TLR agonist comprises dsRNA, poly I:C, poly ICIC, poly IC12U, IPH302, ARNAX and/or MPLA. In some embodiments, the TLR agonist activates TLR4, where the TLR agonist comprises LPS, lipoteichoic acid β-defense peptide 2, fiber binding protein EDA, HMGB1, snapin, tenascin C, OK-432, AS04 and/or GLA-SE. In some embodiments, the TLR agonist activates TLR5, where 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, where 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 a bistriazole base 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, poly I:C and/or R848. In some embodiments, the pro-inflammatory agent comprises 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 interleukin cascade or interleukin 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, 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, as appropriate. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

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 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 (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 SHP-1 inhibitor are separated by two, three, four, five, six, seven, eight, nine or ten days. In some embodiments, each administration of SHP-1 inhibitor is separated by at least one day from the previous or subsequent administration of 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 for at least two consecutive days (e.g., at least three consecutive days) in each cycle. In some embodiments, each cycle has about seven to about twenty days. In some embodiments, the TLR agonist activates TLRs on macrophages, where the TLRs include TLR9 as appropriate. 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 includes CpG, poly I:C, and/or R848. In some embodiments, the proinflammatory agent includes an agent selected from the group consisting of or is selected from the group consisting of: R848, 3M-852A, Motomod, Bropirimine, and Visamod. 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 an analog or derivative thereof 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 interleukin cascade or interleukin 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.

In some embodiments, a method of treating cancer (e.g., a solid tumor, such as a hematological cancer, such as 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 STING activator (e.g., cGAMP, such as 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 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 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 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 nano vaccine (such as PC7A 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 interleukin cascade or interleukin 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, 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, as appropriate. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

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 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 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 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 radiosensitizers and radioprotectants. 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, intracavitary 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 interleukin cascade or interleukin 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, 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, as appropriate. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

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 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, as appropriate, 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 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 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 hour, 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 radiosensitizers and radioprotectants. 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, intracavitary 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 interleukin cascade or interleukin 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, 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, as appropriate. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

In some embodiments, a method of treating cancer (e.g., a solid tumor, such as a hematological cancer, such as an advanced cancer) in an individual is provided, 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 at least twice (which are separated by at least one day), as appropriate. 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, TPI-1, or an analog or derivative thereof, and the radiation treatment are administered on the same day. In some embodiments, TPI-1, or an analog or derivative thereof, and/or the radiation treatment are administered at least two times (e.g., at least three, four, five, or six times). In some embodiments, TPI-1, or an analog or derivative thereof, and the radiation treatment are administered for at least two cycles (e.g., at least three cycles), optionally wherein in each cycle TPI-1, or an analog or derivative thereof, and the radiation treatment are administered on the same day 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 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 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, TPI-1 or an analog or derivative thereof 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 interleukin cascade or interleukin 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.

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 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 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 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 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 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 pro-inflammatory agent is a PAMP activator. In some embodiments, the PAMP activator is a triacyl lipopeptide, LPS, lipoprotein, peptidoglycan, zymosan, lipoteichoic acid, styrax phospholipid, Pam3Cys porin, adipoarabinomannan, double-stranded RNA, poly (I: C), styrax 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 pro-inflammatory agent is a DAMP activator. In some embodiments, the DAMP activator is a preventin, HSP60, HSP70, messenger RNA, low molecular weight hyaluronic acid, fibrinogen, fibronectin, fx1-defensin peptide, 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 interleukin cascade or interleukin 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, 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, as appropriate. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

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, as appropriate, 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 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 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 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, TTI-662, RRx-001, Lanotuzumab (MCS110), LY3022855, SNDX-6352, Emactuzumab (RG7155), Pexidartinib (PLX3397), CAN04, Canakinumab (ACZ885), BMS-986253, Pepinemab (VX15/2503), Trebananib, FP-1305, Enapotamab 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 interleukin cascade or interleukin 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, 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, as appropriate. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

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 proinflammatory interleukin (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, as appropriate, 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 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 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 proinflammatory cytokines promote M1 macrophages. In some embodiments, the proinflammatory cytokines comprise or are TNF, IFNγ, and/or GM-CSF. In some embodiments, the proinflammatory cytokines comprise IFNγ. In some embodiments, the proinflammatory cytokines comprise IL-1. In some embodiments, the proinflammatory cytokines comprise TNF-a. In some embodiments, the proinflammatory cytokines comprise IL-6. In some embodiments, the SHP-1 inhibitor is administered systemically and the proinflammatory cytokines are 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 interleukin cascade or interleukin 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.

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 daily for no more than three or two consecutive days and, as appropriate, 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 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 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, cyclophosphamides, isocyclic phosphamides), 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 anti-metabolite. In some embodiments, the anti-metabolite is selected from the group consisting of: cytidine analogs (e.g., azacitidine, decitabine, cytarabine, gemcitabine), folic acid antagonists (e.g., methotrexate, pemetrexed), purine analogs (e.g., cladribine, clofarabine, nelarabine), pyrimidine analogs (e.g., 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 topoisomerase II inhibitors (e.g., anthracyclines, doxorubicin, daunorubicin, idarubicin, mitoxantrone), topoisomerase I inhibitors (e.g., irinotecan, topotecan), taxanes (e.g., paclitaxel, docetaxel, cabazitaxel), vinca alkaloids In some embodiments, the chemotherapeutic agent is a hydroxyurea, retinoic acid, 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 interleukin cascade or interleukin 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.

In some embodiments, a method of treating cancer (e.g., a solid tumor, such as a hematological cancer, such as 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, such as a hematological cancer, such as 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 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 a SHP-1 inhibitor to a subject 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 SHP-1 inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and/or locally (e.g., intratumorally). In some embodiments, the cancer vaccine comprises a cell-based vaccine, a peptide-based vaccine, a virus-based vaccine, and/or a nucleic acid-based vaccine. In some embodiments, the 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 interleukin cascade or interleukin 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.

In some embodiments, a method of treating cancer (e.g., a solid tumor, such as a hematological cancer, such as 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 an oncolytic virus, wherein the SHP-1 inhibitor is administered at least twice (at least three, four, five, or six times), as appropriate. In some embodiments, a method of treating cancer (e.g., a solid tumor, such as a hematological cancer, such as 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 an oncolytic virus, 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 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 a SHP-1 inhibitor to a subject 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 SHP-1 inhibitor is administered systemically (e.g., intravenously, e.g., subcutaneously) and/or locally (e.g., intratumorally). In some embodiments, the oncolytic virus comprises or is: adenovirus (e.g., ONYX-15, LOAd703 virus), protoparvovirus, parvovirus (e.g., H-1PV), vaccinia virus (VACV), Reovirus (e.g., Reolysin), or herpes simplex virus (HSV, e.g., HSV-1, HSV-2, G207, L1BR1, HF10, T-VEC, Orien X010). In some embodiments, the oncolytic virus comprises JX-593, Coxsackievirus A21 (CVA21), marabá 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 interleukin cascade or interleukin 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, 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, as appropriate. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

In some embodiments, a method of treating cancer (e.g., a solid tumor, such as a hematological cancer, such as 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 ultrasound therapy (e.g., high intensity focused ultrasound (HIFU), such as low intensity focused ultrasound (LIPUS)), 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, such as a hematological cancer, such as 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), such as low intensity focused ultrasound (LIPUS)), wherein the SHP-1 inhibitor and the 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, as appropriate, 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 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 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 sonic 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 interleukin cascade or interleukin 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, 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, as appropriate. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

In some embodiments, a method of treating cancer (e.g., a solid tumor, such as a hematological cancer, such as 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 magnetic therapy (e.g., a pulsed magnetic field, such as 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 the 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 daily for no more than three or two consecutive days and, as appropriate, 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 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 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 interleukin cascade or interleukin 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, 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, as appropriate. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

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 electrotherapy or electrochemical 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 electrical or electrochemical treatment, wherein the SHP-1 inhibitor and the electrical or electrochemical treatment 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, as appropriate, 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 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 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 electrochemical 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 interleukin cascade or interleukin 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, 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, as appropriate. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

In some embodiments, a method of treating cancer (e.g., a solid tumor, such as a hematological cancer, such as 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, such as a hematological cancer, such as 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 the electrostatic therapy are administered within 24 hours (e.g., within 12, 8, 4, 2, or 1 hour, or within 30 minutes) of each other. 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, as appropriate, 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 a SHP-1 inhibitor to a subject 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 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 electrostatic therapy at 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 interleukin cascade or interleukin 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.

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), wherein the subject is selected for treatment based on the subject having a persistent inflammatory response. In some embodiments, the subject has an acute inflammatory response. In some embodiments, the inflammatory response is located in a tumor. In some embodiments, the inflammatory response is located in a different location than the tumor. In some embodiments, the 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 interleukins (e.g., IFNγ, IL-12b, TNFα, IL-6, IL-1b, IFN-a1, IFN-a2, IFN-b1), b) an increase in one or more (e.g., at least one, two, or three) anti-inflammatory interleukins (e.g., TGFb1, Tgfb2, Tgfb3, Tgfb4, Tgfb5, Tgfb6, Tgfb7, Tgfb8, Tgfb9, Tgfb10, Tgfb111, Tgfb12, Tgfb13, Tgfb14, Tgfb15, Tgfb16, Tgfb17, Tgfb18, Tgfb19, Tgfb20, Tgfb19, Tgfb21, Tgfb19, Tgfb22, Tgfb19 ... In some embodiments, the SHP-1 inhibitor is administered intermittently. In some embodiments, the SHP-1 inhibitor is administered for no more than three or two consecutive days and at least twice a 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 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, pseudopteroylxanthrone dimer 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 subject 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 subject 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 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 interleukin cascade or interleukin 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.

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), wherein the subject is selected for treatment based on the subject having persistent immunogenic cell death (ICD). In some embodiments, the subject has ICD when a sample from the cancer has a higher level (e.g., at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%) of one or more DAMPs than a reference sample (e.g., a corresponding sample in 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 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 DAMP is selected from the group consisting of endoplasmic reticulum (ER) chaperones (e.g., calcium reticulum chaperones (CALRs), such as heat shock proteins (HSPs)), non-histone chromatin-bound protein high mobility group box 1 (HMGB1), cytoplasmic protein phospholipid binding protein A1 (ANXA1), and small metabolites ATP, and type I interferons (IFNs). 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, pseudopiperidin dimer 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 subject 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 subject 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 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 interleukin cascade or interleukin 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.

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) a monocyte or macrophage 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 monocyte or macrophage is derived from the same individual. In some embodiments, the monocyte or macrophage is engineered to express a chimeric receptor targeting a tumor antigen. In some embodiments, the monocyte or macrophage and the proinflammatory agent are administered within 24 hours of each other (e.g., within 12, 8, 4, 2, or 1 hour, or within 30 minutes). In some embodiments, monocytes or macrophages and proinflammatory agents are administered simultaneously, concurrently, or sequentially. In some embodiments, monocytes or macrophages are administered before the proinflammatory agent. In some embodiments, 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 interleukin cascade or interleukin 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, 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, as appropriate. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

The present application also provides a method of regulating monocytes or macrophages from an individual suffering from cancer, 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 interleukin cascade or interleukin 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, 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, as appropriate. In some embodiments, the SHP-1 inhibitor comprises TPI-1.

The present application also provides a method of activating phagocytosis of tumor cells in an individual with a tumor, 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 of activating tumor-infiltrating T cells in an individual with a tumor, 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 of no more than once every three days. In some embodiments, the method comprises administering a SHP-1 inhibitor to a subject 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 proinflammatory 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, radiation therapy, a PAMP/DAMP activator, a checkpoint inhibitor, a proinflammatory cytokine, a chemotherapeutic agent, a bacterial component, a cancer vaccine, and an oncolytic virus. In some embodiments, the method further comprises administering to the individual an agent that reduces systemic inflammation and/or reduces the inflammatory interleukin cascade or interleukin 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.

It was also found that the strong inhibitory regulation by iR-SHP-1 in tumors is largely dependent on the physical contact between cancer cells and macrophages. Separation of cancer cells from macrophages using transwells that allow soluble factor delivery but prevent cancer cells from "contacting" macrophages failed to exert a strong inhibitory effect on macrophage proinflammatory responses. See, e.g., FIG. 19 . Combination of inhibitors of iR (e.g., antibodies, fusion proteins, or other agents that block a) SIGLEC interaction with sialoproteoglycans, 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 resulted in abrogation of iR-mediated inhibition, allowing macrophage activation in a pro-inflammatory direction, as evidenced by the increase in interleukins shown in Figure 27C.

Therefore, it is also carefully considered that iR blockers (such as antibodies, fusion proteins or other agents that block the interaction between SIGLEC and sialic acid proteoglycan 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 lymphocyte activation molecule family (SLAMF) receptors and their ligands, etc.), especially combinations of such blockers, can be used to replace the SHP-1 inhibitors in the methods described herein. See, for example, FIG. 27C. In some embodiments, a method of treating cancer is provided, comprising administering at least two or three inhibitors that block different interactions selected from the group consisting of: SIGLEC-sialic acid proteoglycan, LILRB-MHC, CD47-SIRPa, lectin-lectin receptor, signaling lymphocytic activation molecule family (SLAMF) receptor-its ligand; and optionally a pro-inflammatory agent.

Blockers described herein include any agent that: a) is capable of reducing the binding of an inhibitory receptor and its ligand, as measured by, for example, spectroscopic analysis, isothermal titration calorimetry (ITC), optical biosensors such as 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.

In some embodiments, the method comprises administering to a subject in need thereof: a) a CD47-SIRPα blocker (e.g., an anti-CD47 antibody (e.g., B6H12) or an anti-SIRPα antibody), b) a LILRB-MHC blocker (e.g., an antibody against LILRB1, LILRB2 and/or LILRB3, such as an antibody against HLA-A, HLA-B and/or HLA-C), c) a SIGLEC-sialic acid proteoglycan blocker (e.g., anti-siglec9, anti-siglec7, anti-siglec8, such as neuraminidase), and optionally d) a pro-inflammatory agent (e.g., a TLR agonist, a STING activator). In some embodiments, the method comprises administering to a subject in need thereof: a) neuraminidase, b) anti-CD47 antibody, c) anti-HLA-A/B/C, and optionally d) a pro-inflammatory agent (e.g., TLR agonist, STING activator). Exemplary antibodies against iR or its ligands can be seen in FIG. 27B. In some embodiments, the subject has a persistent infection and does not require treatment with a pro-inflammatory agent. Tumor Microenvironment (TME) Immunosuppression and SHP-1 Signaling

Src homology region 2 (SH-2) domain-containing phosphatase 1 (SHP-1) is a non-receptor tyrosine phosphatase encoded by the PTPN6 gene located on human chromosome 12p13 and containing two promoter regions (in exons 1 and 2), which 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 transcripts of SHP-1. The two SHP-1 isoforms show different subcellular localizations: form I is primarily localized in the nucleus, whereas form II is localized in the cytoplasm, suggesting that they have different targets.

SHP-1 is a 595 amino acid protein composed of two tandem N-terminal SH2 domains (N-SH2 and C-SH2), one is a typical catalytic protein tyrosine phosphatase (PTP) domain, and one is a C-terminal tail containing several phosphorylation sites. Its crystals show the following structure: In which N-SH2 binds to the catalytic site of the protein via charge-charge interactions. In this autoinhibited inactive state, substrate is prevented from approaching the active site, but binding of the phosphotyrosine residue to the SH2 domain causes a conformational change that weakens the interaction between N-SH2 and the catalytic domain. This opens the conformation to allow substrate access and is further stabilized by new interactions between the SH2 domain and the catalytic domain. These molecular rearrangements determine a complex regulatory mechanism controlled by substrate recruitment.

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) residues. Tyr536 and Tyr564 become phosphorylated following a variety of 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 an interaction with the N-SH2 domain, thereby releasing the inhibitory effect of this domain 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, the mechanism of which has not been fully characterized.

Protein-tyrosine phosphorylation is a reversible post-translational modification 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 serves as a classical tumor suppressor, primarily involved in the in vivo homeostatic maintenance of potentially all these processes. SHP-1 function is actually altered in solid and hematological human cancers through somatic mutations or epigenetic mechanisms. In addition to its well-documented role in regulating hematopoietic cell biology, SHP-1 has been implicated in multiple signaling pathways involved in cancer pathogenesis and progression.

However, inhibition of SHP-1 causes severe side effects. Studies of SHP-1-deficient mice (me/me or me ^v /me ^v ) have revealed key immunological abnormalities and overactivation of immune cells associated with global loss of SHP-1 (6, 7). Meat-depleted mice typically die at an early age from life-threatening autoimmune inflammatory conditions. Even partial depletion of SHP-1 in wild-type mice causes features of inflammatory disease as they grow into 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, interleukin storms, and enhanced autoimmunity.

Inhibitors targeting SHP-1 phosphatase activity have been in development for some time, and several are now in preclinical studies, including NSC-87877, sodium antimony gluconate (SSG), tyrosine phosphatase inhibitor 1 (TPI-1 or its analogs or derivatives), and suramine; 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 malignant tumors (NCT00629200); the drug is administered in combination with interferons, 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 inhibitors are in Phase II trials. Agents that reduce systemic inflammation

In some cases, individuals develop systemic inflammation, i.e., cytokine release syndrome (CRS), after receiving, for example, immunotherapeutic treatment, although the inflammatory disorder is not fully understood. CRS may be induced by direct targeted cell lysis and continuous release of cytokines (such as TNFα or IFNγ) or by T cell activation due to therapeutic stimulation followed by subsequent cytokine release. These cytokines trigger a chain reaction due to activation of innate immune cells (such as macrophages and endothelial cells), which then induces further cytokine release. Specifically, IL6, IL10, and IFNγ are most commonly found to be elevated in patients with CRS.

The methods described herein may further comprise administering an agent that reduces systemic inflammation (including, for example, an agent that reduces the inflammatory interleukin cascade or interleukin storm) so as to inhibit 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 (e.g., before or after) with the SHP-1 inhibitor. In some embodiments, the agent that reduces systemic inflammation is administered on the same dosing schedule as the SHP-1 inhibitor. In some embodiments, the agent that reduces systemic inflammation is administered in a subtherapeutic dose, i.e., in an amount 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.).

The agent may include any anti-inflammatory agent known in the art, including inhibitors of proinflammatory agents or antagonists of proinflammatory agents. For example, the agent 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 agents that target one or more molecules (e.g., by inhibiting TNFα alone, IL6 alone, or a combination of TNFα and IL6). Anti- TNFα Antagonists

The major pro-inflammatory interleukin TNFα is secreted by activated macrophages, monocytes and lymphocytes. The inventors unexpectedly discovered that administration of an anti-TNFα antibody to an individual who has been administered a SHP-1 inhibitor and a pro-inflammatory agent alleviates toxicity caused by systemic inflammation without compromising the efficacy of the therapeutic agent.

Thus, in some embodiments, the methods of the present application comprise 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.

Antibodies suitable for use in the methods provided herein include, but are not limited to: Remicade® (Infliximab (Centocor)); and those antibodies described, for example, in U.S. Patent Nos. 6,835,823, 6,790,444, 6,284,471, 6,277,969, 5,919,452, 5,698,195, 5,656,272, and 5,223,395, and European Patent No. 0610201, the contents of each of which are incorporated herein by reference in their entirety; or antibodies that bind to the same antigenic determinant as Remicade®. 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 incorporated herein by reference in their entireties; Simponi™ (Golimimab, CNTO 148 (Centocor)), as described in PCT Publication No. WO 02/12502 and related patents and applications, the contents of which are incorporated herein by reference in their entireties; ART621 (Arana Therapeutics), SSS 07 (Epitopmics and 3SBio) or antibodies that bind to the same epitope as Humira, Simponi, ART621 or SSS 07.

In some embodiments, the TNFα inhibitor (e.g., 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 the following: U.S. Patent No. 5,712,155, PCT Publication No. WO 91/03553 and related patents and applications, the contents of which are incorporated herein by reference in their entirety.

In some embodiments, the TNFα inhibitor (eg, anti-TNFα antagonist) is a modified antibody antagonist or a non-antibody based antagonist. Such antagonists include advanced antibody therapies, such as antibody fragments, including but not limited to Cimzia™ (Certolizumab pegol, CDP870 (Enzon)), bispecific antibodies, Nanobodies® (such as ABX 0402 (Ablynx)), immunotoxins and radiolabeled therapeutics; peptide therapeutics; gene therapies, 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 incorporated herein by reference in their entirety.

In some embodiments, a 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 a 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, a TNFα inhibitor (e.g., an anti-TNFα antibody) is administered concurrently or simultaneously with a SHP-1 inhibitor and/or a proinflammatory agent. In some embodiments, a TNFα inhibitor (e.g., an anti-TNFα antibody) is administered immediately (e.g., within 1 hour or 30 minutes) after administration of a SHP-1 inhibitor and/or a proinflammatory agent.

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 interleukin release syndrome or proinflammatory organ damage. In some embodiments, administration of the TNFα inhibitor does not impair or minimally impairs tumor clearance. Anti- IL6 Antagonists

"Anti-IL6 antagonists" or "IL6 inhibitors" refer to agents that inhibit or block the biological activity of IL6 by binding to IL6 or an IL6 receptor. In some embodiments, the anti-IL6 antagonist is an antibody. In one embodiment, the anti-IL6 antagonist is an antibody that binds to an IL6 receptor. Antibodies that bind to an IL-6 receptor include tocilizumab (including its intravenous i.v. and subcutaneous s.c. formulations) (Chugai, Roche, Genentech), satralizumab (Chugai, Roche, Genentech), sarilumab (Sanofi, Regeneron), NI-1201 (Novimmune and Tiziana), and vobarilizumab (Ablynx). In one embodiment, the anti-IL6 antagonist is a monoclonal antibody that binds 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.

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 every day. In some embodiments, the IL6 inhibitor is administered intermittently. In some embodiments, the IL6 inhibitor is administered to the individual for at least two cycles, wherein each cycle has about three to about seven days. SHP-1 inhibitors

SHP-1 inhibitors referred to herein are any type or class of agents that inhibit 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.

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

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), protein agents containing SH2 domains (which inhibit SHP-1 activation by competing with ITIM motifs), and tyrosine kinase inhibitors that inhibit ITIM phosphorylation.

In some embodiments, a SHP-1 inhibitor does not significantly inhibit SHP-2 (e.g., does not inhibit SHP-2 activity by more than 50%, 40%, 30%, or 20%).

In some embodiments, a SHP-1 inhibitor also inhibits SHP-2.

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

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

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

In some embodiments, the SHP-1 inhibitor is a competitive inhibitor. In some embodiments, the SHP-1 inhibitor is pseudostem oxanthrone dimer A (PXA) or pseudostem oxanthrone dimer B (PXB). See, e.g., Yang et al., ACS Omega. 2020 Sep 29;5(40):25927-25935.

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 binds to the catalytic site). 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 assessed using a variety of methods, such as the rapid SHP-1 PTP assay. See "Materials and Methods" in Kundu et al.

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

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, FIG. 6 of Kundu et al.

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

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

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).

In some embodiments, the SHP-1 inhibitor is pseudo-piperidinoxantrone dimer A (PXA).

In some embodiments, the SHP-1 inhibitor is a PKCθ activator (such as PMA).

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

In some embodiments, the SHP-1 inhibitor is a dominant negative SHP-1 or a constitutively active SHP-1 mutant. See, e.g., WO2009/023333.

In some embodiments, the SHP-1 inhibitor is a nucleic acid editing system (such as 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 genome of the cell. In this approach, the CRISPR system only cleaves the genome DNA of the cell for a limited period of time. See, e.g., Fister et al., Front Plant Sci. 2018 Mar 2;9:268.

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.

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

In some embodiments, the method comprises administering a 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.

In some embodiments, the method comprises administering a 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.

In some embodiments, the method comprises administering a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) for at least two cycles. In some embodiments, a 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).

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 systemically and locally (e.g., intratumorally).

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

In some embodiments, a SHP-1 inhibitor modulates monocytes or macrophages in vitro (e.g., monocytes or macrophages derived from an individual to be treated).

In some embodiments, the SHP-1 inhibitor and the proinflammatory agent described below 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 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. Proinflammatory Agents

Infection and tissue damage are two typical instigators of inflammation. See, for example, Medzhitov, Nature. 2008 Jul 24;454(7203):428-35. Proinflammatory agents as described herein include at least two overlapping categories: 1) any class or type of agent or therapy that can promote inflammation (e.g., by promoting one or more proinflammatory cytokines or interleukins, inhibiting one or more anti-inflammatory cytokines or interleukins, 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 (such as regulatory T cells or MDSCs)), and 2) agents or therapies that can cause cancer cell damage (e.g., cancer cell necrosis).

In some embodiments, the proinflammatory agent triggers proinflammatory signals on macrophages. See, e.g., FIG. 5A . In some embodiments, the proinflammatory agent activates TLRs, TNFRs, or ITAM-Rs. 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, TLR agonists that directly activate TLRs on macrophages, or radiation therapy that indirectly activates proinflammatory signals on macrophages, both exhibit significant anti-tumor effects when used with a SHP-1 inhibitor. See Examples.

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

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 or chemokine, chemotherapy, a bacterial component, a cancer vaccine, an oncolytic virus, sound wave therapy (e.g., high-intensity focused ultrasound), magnetic therapy, electrical therapy, and electrostatic therapy.

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, sound wave therapy (e.g., high-intensity focused ultrasound), magnetic therapy, electrical therapy, and electrostatic therapy.

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

In some embodiments, the pro-inflammatory agent is magnetic therapy (e.g., a pulsed magnetic field, e.g., 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.

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

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

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

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.

In some embodiments, the pro-inflammatory agent comprises a TLR agonist (eg, R848) and an interleukin (eg, IFN -γ).

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

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 HSP, HMGB1, uric acid, fibronectin, and other extracellular matrix proteins. It has also been shown that TLR1 and TLR6 can heterodimerize with TLR10; however, the TLR agonists recognized by this dimer remain to be identified. TLR3 recognizes viral dsRNA, as well as synthetic analogs of dsRNA, such as the ligand poly I:C. TLR4 binds LPS in a complex with lipid A-bound protein CD14 and myeloid differentiation protein 2 (MD2) and recognizes a variety of DAMPs. Endogenous TLR4 ligands that have been described include β-defense peptide 2, fibronectin extradomain 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 of the highly conserved TIR domain. TLR10 is a surface receptor whose natural ligand is still unknown. TLR11, -12, and -13 are present in mice but not in humans. TLR11 was shown to bind to Toxoplasma gondii profilin and Escherichia coli. The ligand for TLR12 has not yet been identified, and TLR13 is an endosomal receptor that recognizes VSV. See, e.g., Kaczanowska et al., J Leukoc Biol. 2013 Jun;93(6):847-63.

TLR signaling can be used as a double-edged sword in cancer. TLR stimulation of cancer cells has been found to lead to tumor progression or inhibition. For example, stimulation of TLR 2, 4, and 7/8 has been found to cause tumor progression via the production of immunosuppressive cytokines, increased cell proliferation, and resistance to apoptosis. R848-stimulation of pancreatic cancer cell lines overexpressing TLR7/8 caused increased cell proliferation and decreased chemosensitivity. On the other hand, stimulation of TLR 2, 3, 4, 5, 7/8, and 9, often combined with chemotherapy or immunotherapy, can lead to tumor inhibition via different pathways. See, e.g., Grimmig et al., Int J Oncol. (2015) 47:857-66; Urban-Wojciuk et al., Front Immunol. 2019; 10: 2388.

In some embodiments, a TLR agonist activates any one of the TLRs.

In some embodiments, the TLR agonist activates TLR1 or TLR2, where appropriate, wherein the TLR agonist comprises triacylated lipoprotein, peptidoglycan, zymosan and/or Pam ₃ CSK ₄ .

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, a fiber binding protein, and/or an ECM protein.

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

In some embodiments, the TLR agonist activates TLR3, optionally wherein the TLR agonist comprises dsRNA, poly I:C, poly ICIC, poly IC12U, IPH302, ARNAX and/or MPLA.

In some embodiments, the TLR agonist activates TLR4, optionally wherein the TLR agonist comprises LPS, lipoteichoic acid β-defense peptide 2, fiber binding protein 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 a bistriazole group 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 Pam ₃ CSK ₄ .

In some embodiments, the TLR agonist activates TLR11, optionally wherein the TLR agonist comprises Toxoplasma gondii profibromin.

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 a TLR on a macrophage.

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

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

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

In some embodiments, the TLR agonist comprises CpG. In some embodiments, the TLR agonist comprises poly I:C. In some embodiments, the TLR agonist comprises CpG and/or poly I:C. In some embodiments, the TLR agonist comprises CpG, poly I:C and/or R848.

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

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 is avoided. Radiation therapy

In some embodiments, the proinflammatory agent comprises or is radiation therapy. Radiation activates an interconnected network of interleukins, 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, e.g., McKelvey et al., Mamm Genome. 2018; 29(11): 843-865.

In some embodiments, radiation therapy comprises irradiating the area of the cancer to be treated.

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

In some embodiments, the radiation therapy is intraoperative radiation therapy ("IORT"). In specific embodiments, the radiation is limited to the tumor site. The patient may undergo intraoperative radiation before or after the tumor is removed. 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 radiation therapy ("SRS").

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, radiation therapy can include subjecting the patient to one or more types of radiation therapy.

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

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

In some embodiments, the radiation therapy is external beam radiation therapy, which may include 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, radiation therapy comprises administering a radiopharmaceutical. The radiopharmaceutical can be delivered via any medium, such as a cell, protein, or small molecule complex. In some embodiments, the radiopharmaceutical is administered to tumor tissue. See, e.g., Sgouros et al. Radiopharmaceutical therapy in cancer: clinical advances and challenges. Nat Rev Drug Discov 19, 589-608 (2020).

In some embodiments, the radiation therapy is brachytherapy, which optionally includes interstitial brachytherapy, intracavitary brachytherapy, intracavitary radiation therapy, and intravenously administered radiolabeled molecules. STING activators

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

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). After 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 dramatic conformational change in the C-terminal domain of STING, mediating its transport to the Golgi compartment. At the Golgi, STING recruits TANK-binding kinase 1 (TBK1), which promotes IRF3 phosphorylation, nuclear translocation, and potently induces transcription of type I IFNs (e.g., IFN-β). STING also triggers a robust proinflammatory cytokine response [e.g., tumor necrosis factor (TNF)] by activating nuclear factor-κB (NF-κB), and this part of the pathway can 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.

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

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

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

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).

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

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

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. PAMP/DAMP Activators

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

Organisms sense microbial infection via innate receptors encoded in their genomes, called pattern recognition receptors, including Toll-like receptors (TLRs), nucleotide- and oligomerization domain-binding (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), which are molecules released by sterile injury. Thus, PAMPs and DAMPs that bind to the same type of receptor initiate the same intracellular pathway and terminate in the same effector function. See, e.g., Alisi et al., Hepatology. 2011 Nov;54(5):1500-2.

In some embodiments, the proinflammatory agent is a PAMP activator. Exemplary PAMP activators include triacyl lipopeptides, LPS, lipoproteins, peptidoglycan, zymosan, lipoteichoic acid, lecithin, Pam3Cys porin, lipoarabinomannan, double-stranded RNA, poly (I: C), lecithin 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 LPS.

In some embodiments, the proinflammatory agent is a DAMP activator. Exemplary DAMP activators include defense peptides, HSP60, HSP70, messenger RNA, low molecular weight hyaluronic acid, fibrinogen, fiber binding protein, fx1-defense peptide, heparan sulfate, HSP60, HSP70, HSP90, HMGB1, and unmethylated CpG DNA. Chemotherapeutic agents

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

In some embodiments, the chemotherapeutic agent is an alkylating agent. Exemplary alkylating agents include nitrogen mustards (e.g., endamustine, cyclophosphamide, isocyclophosphamide), 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).

In some embodiments, the chemotherapeutic agent is an anti-metabolite. Exemplary anti-metabolites 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 (prodrug of 5-FU)).

In some embodiments, the chemotherapeutic agent is an anti-microtubule agent. Exemplary anti-microtubule agents include topoisomerase II inhibitors (e.g., anthracycline, doxorubicin, daunomycin, idamycin, 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).

Other exemplary chemotherapeutic agents include hydroxyurea, retinoic acid, arsenic trioxide, and proteasome inhibitors (e.g., bortezomib). Proinflammatory interleukins

In some embodiments, the proinflammatory agent is a proinflammatory cytokine.

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

In some embodiments, the proinflammatory interleukins include IL-6, TNFα, interleukins from the IL-1 family (eg, IL-1α, IL-1β, IL-18, IL-33, and IL-36), and/or IFNγ.

In some embodiments, the proinflammatory cytokines include cytokines from the IL-1 family. In some embodiments, the proinflammatory cytokines include 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). Checkpoint inhibitors

In some embodiments, the proinflammatory agent is a checkpoint inhibitor. 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), planned cell death receptor-1 (PD-1), and planned cell death ligand-1 (PD-L1). See, e.g., Marin-Acevedo et al., J Hematol Oncol 14, 45 (2021).

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

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: ripimumab, cemiprilizumab, 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), ALX148, TTI-662, RRx-001, larotuzumab (MCS110), LY3022855, SNDX-6352, emeticholic acid (RG7155), pirituximab (PLX3397), CAN04, canakinumab (ACZ885), BMS-986253, pebinetuzumab (VX15/2503), teberanib, FP-1305, empatuzumab vedotin (EnaV), or bavituximab. Cancer vaccines

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

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

Cell-based vaccines were originally a form of cancer vaccine. Cell-based cancer vaccines are usually prepared from whole cells or cell fragments, contain almost all tumor antigens, and induce a broad 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 clinical practice. Viruses are naturally immunogenic and their genetic material can be engineered to contain sequences encoding tumor antigens. Some recombinant viruses (such as adenoviruses) can be used as vectors to infect immune cells. Engineered viral vaccines can present large amounts of tumor antigens in the immune system and produce anti-tumor immunity. In addition, oncolytic viruses can also be used as vectors. In addition to providing tumor antigens, the virus itself can also dissolve tumors and release tumor antigens, further increasing the effectiveness of the vaccine and producing long-term immune memory.

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

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

Nucleic acid vaccines induce potent MHC I-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 antigenic determinants or present several antigens simultaneously. Finally, nucleic acid vaccine preparations are simple and rapid, which makes them suitable for the development of personalized neoantigen cancer vaccines. Oncolytic viruses

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

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, e.g., Cancer Cell. 2022 Aug 15;S1535-6108(22)00357-9.

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

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

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 a 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. Immune cell, monocyte or macrophage

The immune cells described in this article cover a wide variety of immune cells.

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, 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.

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 does not exceed about 10, 9, 8, 7, 6, 5, 4, or 3 days.

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

Methods for engineering monocytes or macrophages to temporarily express reduced levels of SHP-1 are well known in the art. Exemplary methods include contacting monocytes or macrophages with SHP-1 inhibitors described herein (such as 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 vivo or in vitro.

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 cell comprises a neutrophil (e.g., a neutrophil expressing a CAR).

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

In some embodiments, the immune cells are derived from the same individual (i.e., autologous). In some embodiments, the immune cells are allogeneic.

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 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).

In some embodiments, the immune cells express one or more pro-inflammatory cytokines, optionally wherein the one or more pro-inflammatory 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 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.

In some embodiments, the SHP-1 inhibitor, immune cells, and proinflammatory agents 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, immune cells are administered simultaneously or concurrently with the SHP-1 inhibitor and/or proinflammatory agent. Inflammatory response or persistent infection

There is a large body of evidence that both acute and chronic inflammation are involved in the development and progression of cancer. Advances in the study of inflammation have shown a relationship between the inflammatory process and the neoplastic transformation, the progression of tumors, and the development of metastasis and recurrence. In addition, tumor invasive procedures (both surgery and biopsy) affect the remaining tumor cells by increasing their survival, proliferation and migration. One of the concepts that explains this phenomenon is the induction of wound healing responses. While in normal tissues it is essential for tissue repair, in tumor tissues, the induction of acquired and innate immune responses associated with wound healing stimulates tumor cell survival, angiogenesis and extravasation of circulating tumor cells. See, e.g., Singh et al. Ann Afr Med. 2019 Jul-Sep; 18(3): 121-126; Piotrowski et al. Rep Pract Oncol Radiother. 2020 May-Jun; 25(3): 422-427.

However, as demonstrated in this application, the combination of a SHP-1 inhibitor and a proinflammatory agent unleashes a proinflammatory response and transforms the immunosuppressive tumor environment into a place with inflammatory hallmarks. See, e.g., FIG. 7F . A significant anti-tumor effect was achieved. These results provide support for the use of the methods described herein to treat individuals in an inflammatory response.

In some embodiments, when treated with the methods described herein, the subject is in an inflammatory response or has an ongoing infection. 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 interleukins (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 interleukins (e.g., TGFb1, TGFb2, TGFb3), c) d) increase in infiltrating immune cells (such as T cells, NK cells, macrophages, neutrophils), d) decrease in suppressive immune cells (such as MDSCs), and/or e) increase in one or more (such as at least one, two, three, four or five) immunogenic co-stimulatory molecules (such as CD80, CD86, OX40L, CD40, ICOS-L, PD-L1, GITRL) in tissues (such as tumor tissues) or immune cells (such as macrophages).

In some embodiments, the inflammatory response is an acute inflammatory response.

In some embodiments, the inflammatory response is located in a tumor. In some embodiments, the inflammatory response is located in a different location than the tumor.

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 interleukins (e.g., IFNγ, IL-12b, TNFα, IL-6, IL-1b, IFN-a1, IFN-a2, IFN-b1), b) an increase in one or more (e.g., at least one, two, or three) anti-inflammatory interleukins (e.g., TGFb1, Tgfb2, Tgfb3, Tgfb4, Tgfb5, Tgfb6, Tgfb7, Tgfb8, Tgfb9, Tgfb10, Tgfb111, Tgfb12, Tgfb13, Tgfb14, Tgfb15, Tgfb16, Tgfb17, Tgfb18, Tgfb19, Tgfb20, Tgfb19, Tgfb21, Tgfb19, Tgfb22, Tgfb19 ... c) decreased TGFb2, TGFb3), c) increased infiltrating immune cells (such as T cells, NK cells, macrophages, neutrophils), d) decreased suppressor immune cells (such as MDSCs), and/or e) increased one or more (such as at least one, two, three, four or five) immunogenic co-stimulatory molecules (such as CD80, CD86, OX40L, CD40, ICOS-L, PD-L1, GITRL) in tissues (such as tumor tissues) or immune cells (such as macrophages).

In some embodiments, the increase described herein refers to a level that is at least about 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, or 200% higher than a reference state, where the reference state is when the subject is neither treated with the methods described herein nor infected with a pathogen. In some embodiments, the increase described herein refers to a level 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, where the reference state is when the subject 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.

In some embodiments, the reduction described herein refers to a level 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, where 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.

In some embodiments, the subject has an inflammatory response (e.g., in a tumor, e.g., in a site different from the tumor) within about one week, 6 days, 5 days, 4 days, 3 days, 2 days, or one day before and/or after administration of the SHP-1 inhibitor.

In some embodiments, the subject has a persistent inflammatory response (e.g., in a tumor, e.g., in a site different from the tumor) when a SHP-1 inhibitor is administered.

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

In some embodiments, the individual has an ongoing infection (e.g., bacterial infection, viral infection, fungal infection), and the method further comprises administering an antibacterial therapy (e.g., antibiotics), an antiviral therapy, an antimicrobial therapy, or an antiprotozoal therapy. Immunogenic cell death

In some embodiments, a subject has immunogenic cell death when treated with the methods described herein.

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 chemotherapeutics, such as anthracyclines, DNA-damaging agents, and proteasome inhibitors; (3) targeted anticancer agents, such as the tyrosine kinase inhibitor crizotinib, the epidermal growth factor receptor-specific monoclonal antibody cetuximab, and poly (ADP-ribose) polymerase (PARP) inhibitors; and (4) various physical modalities, including hypericin-based and remdaporfin-based photodynamic therapy, in vitro photochemotherapy, various forms of ionizing radiation, high hydrostatic pressure, and severe heat shock. It involves the activation of the immune system against cancer in immunocompetent hosts. ICD involves the release of damage-associated molecular patterns (DAMPs) from dying tumor cells, which lead to the activation of tumor-specific immune responses, thus resulting in long-term efficacy of anticancer drugs by combining direct cancer cell killing and anti-tumor immunity. DAMPs include cell surface exposure of calcium chaperones (CRTs) 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, e.g., Ahmed et al., Mol Oncol. 2020 Dec;14(12):2994-3006 and Fucikova et al., Cell Death Dis. 2020 Nov 26;11(11):1013.

Key DAMPs of cell death perceived as immunogenic include calcitonin, high mobility group box 1 (HMGB1), ATP, phospholipid binding protein 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 based on a variety of different pathways. See, e.g., Cell Death Dis. 2020 Nov 26;11(11):1013.

In some embodiments, the subject has an ICD (e.g., in a tumor, e.g., in a different site from the tumor) within about one week, 6 days, 5 days, 4 days, 3 days, 2 days, or one day before and/or after administration of the SHP-1 inhibitor.

In some embodiments, the subject has persistent ICD (e.g., in a tumor, e.g., in a site different from the tumor) when a SHP-1 inhibitor is administered.

In some embodiments, an individual has ICD when a sample from a cancer has a higher level of one or more (e.g., at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% more) DAMPs than a reference sample (e.g., a corresponding sample in a healthy control, e.g., a sample from a cancer before administration of a therapy that induces ICD). In some embodiments, the DAMP is selected from the group consisting of: endoplasmic reticulum (ER) chaperones (e.g., calcitonin chaperones (CALRs), e.g., heat shock proteins (HSPs)), non-histone chromatin-bound protein high mobility group box 1 (HMGB1), cytoplasmic protein phospholipid-bound protein A1 (ANXA1), and small metabolites ATP, and type I interferons (IFNs). Individual

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

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

In some embodiments, the subject has received a therapy that induces an inflammatory response or immunogenic cell death (e.g., radiation therapy) (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 the SHP-1 inhibitor). In some embodiments, the subject is to receive a therapy that induces an inflammatory response or immunogenic cell death (e.g., radiation therapy) (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).

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

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

In some embodiments, the individual is female. In some embodiments, the individual is male.

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

In some embodiments, a subject is selected for treatment based on high expression and/or high activation of SHP-1 in tumor tissue. In some embodiments, a subject has high expression and/or high activation of SHP-1 when the expression and/or activation is at least about 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, or 200% greater than a reference expression and/or activation 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).

In some embodiments, the subject is at risk for developing systemic inflammation and/or CRS. In some embodiments, the subject develops systemic inflammation and/or CRS prior to administration of an agent that reduces systemic inflammation. Interleukin 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, the lungs, kidneys, liver, brain, heart, spleen, or any combination thereof, such as multiple organ failure.

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 interleukin 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 such as corticosteroids. Mild CRS typically resolves within one to two weeks and does not require or necessitate hospitalization.

In some embodiments, the individual does not develop severe interleukin release syndrome. In some embodiments, the individual does not develop grade 2 CRS. In some embodiments, the individual does not develop grade 3 CRS. In some embodiments, the individual does not develop grade 4 CRS. More severe cases are characterized by hypotension and hyperthermia, and severe CRS can progress to an uncontrolled systemic inflammatory response with circulatory shock requiring vasopressors, vascular leak, disseminated intravascular coagulation, and multi-organ system failure. More severe cases of CRS typically require hospitalization. Common laboratory abnormalities in patients with CRS include cytopenias, elevated creatinine and liver enzymes, distorted coagulation parameters, and high CRP. There are four grading systems currently used for interleukin 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 entirety.

In some embodiments, prior to administration of 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, an agent that reduces systemic inflammation improves, eliminates, or reverses CRS, including organ damage, such as proinflammatory organ damage (e.g., nephritis, hepatitis, pneumonia, myocarditis, appendicitis). Table 1. Medical grading system for interleukin release syndrome. Penn grading scale CTCAE v4.0 2014 Lee et al. MDACC Level 1 Mild reactions: Treat with supportive care (such as antipyretics and antiemetics) Mild reaction; infusion interruption not indicated; intervention not indicated Symptoms that are not life-threatening and require only symptomatic treatment, such as fever, nausea, fatigue, headache, myalgia, and malaise Temperature ≥38°C (fever) or Grade 1 organ toxicity Level 2 Moderate reaction: Some signs of organ dysfunction (e.g., grade 2 creatinine or grade 3 LFTs) related to CRS and not attributable to any other condition. Hospitalization for management of CRS-related symptoms, including fever with associated neutropenia and requiring IV therapy (excluding fluid resuscitation for hypotension) Indicated therapy or infusion interrupted but responds promptly to symptomatic treatment (e.g., antihistamines, NSAIDs, anesthetics, IV fluids); indicated prophylactic medication ≤ 24 h Symptoms requiring and responsive to moderate intervention. Oxygen requirement <40%, or hypotension responsive to fluids or low-dose vasopressors, or grade 2 organ toxicity Systolic blood pressure <90 mmHg (hypotension) but responsive to IV fluids or low-dose vasopressors or requiring oxygen (FiO2 <40%), SaO2 >90% (hypoxia); or grade 2 organ toxicity Level 3 More severe reactions: requiring hospitalization for control of symptoms associated with organ dysfunction (including grade 4 LFTs or grade 3 creatinine associated with CRS) that are not attributable to any other condition; this excludes control of fever or myalgias; includes hypotension treated with IV fluids (defined as multiple fluid boluses for blood pressure support) or low-dose vasopressors, coagulopathy requiring fresh frozen plasma or cryoprecipitate or fibrinogen concentrate, and hypoxia requiring supplemental oxygen (nasal cannula oxygen, high-flow oxygen, CPAP, or BiPAP). Patients hospitalized for control of suspected infection due to fever and/or neutropenia may have grade 2 CRS Prolonged response (e.g., failure to respond promptly to symptomatic medications and/or brief interruption of infusion); recurrence of symptoms after initial improvement; indication for other clinical sequelae (e.g., renal injury, pulmonary infiltrates) Symptoms requiring and responding to active intervention. Oxygen requirement ≥40%, or hypotension requiring high-dose or multiple vasopressors, or Grade 3 organ toxicity, or Grade 4 transaminase elevations Systolic blood pressure <90 mmHg (hypotension) requiring high-dose or multiple vasopressors or requiring oxygen (FiO2 ≥40%), SaO2 >90% (hypoxia); or Grade 3 organ toxicity; or Grade 4 transaminase elevation Level 4 Life-threatening complications, such as hypotension requiring high-dose vasopressors and hypoxia requiring mechanical ventilation Life-threatening outcome; vasopressors or ventilator support indicated Life-threatening symptoms. Requirement of ventilator support or grade 4 oxygen toxicity (excluding elevated transaminases) Life-threatening hypotension or requiring mechanical ventilation or grade 4 oxygen toxicity (excluding elevated transaminases)

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

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

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.

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.

Examples of cancers described herein include, but are not limited to, adrenocortical carcinoma, unexplained myeloid metaplasia, AIDS-related cancers (e.g., AIDS-related lymphoma), anal cancer, coccygeal cancer, astrocytoma (e.g., cerebellar and brain), basal cell carcinoma, bile duct carcinoma (e.g., extrahepatic), bladder cancer, bone cancer (osteosarcoma and malignant fibromyoma), brain tumors (e.g., neuroglioma, brain stem neuroglioma, cerebellar or brain astrocytoma (e.g., pilocytic astrocytoma, diffuse astrocytoma, degenerative (malignant) astrocytoma), malignant neuroglioma, ventricular Tubuloma, oligodenglioma, meningioma, craniophysoma, hemangioblastoma, medulloblastoma, supratentorial primitive neuroectodermal tumor, optic pathway and hypothalamic neuroglioma and neuroglioma), 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 tumor (Ewing's family of tumor), eye cancer (such as intraocular melanoma and retinoblastoma), gallbladder cancer, gastric (gastric/stomach) cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor tumor; GIST), germ cell tumors (e.g., extracranial, extragonadal, ovarian), gestational trophoblastic tumor, head and neck cancer, hepatocellular (liver) cancer (e.g., hepatoma and heptoma), hypopharyngeal cancer, islet cell cancer (endocrine pancreas), laryngeal cancer, leukemia, lip and oral cancer, oral cancer, liver cancer, lung cancer (e.g., small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma, and lung squamous 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 and sinus cancer , nasopharyngeal carcinoma, neuroblastoma, neuroendocrine cancer, oropharyngeal cancer, ovarian cancer (e.g., ovarian epithelial carcinoma, ovarian germ cell tumor, ovarian low-grade malignant tumor), pancreatic cancer, parathyroid cancer, penile cancer, peritoneal cancer, pharyngeal cancer, pheochromocytoma, pinealoblastoma and supratentorial primitive neuroectodermal tumor, pituitary tumor, pleural Pulmonary blastoma, lymphoma, primary central nervous system lymphoma (small neuroblastoma), pulmonary lymphangioleiomyomatosis, rectal cancer, kidney cancer, renal pelvis and ureter cancer (transitional cell carcinoma), rhabdomyosarcoma, salivary gland cancer, skin cancer (e.g. non-melanoma (e.g. squamous cell carcinoma), melanoma and Merkel cell carcinoma) cell carcinoma), small intestinal 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-transplant lymphoproliferative disorder (PTLD), abnormal blood vessel proliferation associated with macular degeneration, edema (such as that associated with brain tumors), and Meigs' syndrome.

In some embodiments, cancer is a cancer associated with viral infection. In some embodiments, cancer is a 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 a human herpesvirus 8 (HHV8)-related cancer (e.g., Kaposi sarcoma). In some embodiments, cancer is a human T-lymphotropic virus (HTLV-1)-related cancer (e.g., adult T-cell leukemia or lymphoma). In some embodiments, cancer is an Epstein-Barr virus (EBV)-related cancer (e.g., Burkitt lymphoma, Hodgkin's and non-Hodgkin's lymphoma, gastric cancer). In some embodiments, the cancer is a hepatitis B virus (HBV)-related cancer (e.g., liver cancer). In some embodiments, the cancer is a hepatitis C virus-related cancer (e.g., liver cancer, non-Hodgkin's lymphoma).

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.

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

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

In some embodiments, the cancer is refractory to one or more of radiation therapy, chemotherapy, or immunotherapy (e.g., checkpoint blockade). Administration, Methods of Administration, and Delivery Vehicles

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

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

The proinflammatory agent, SHP-1 inhibitor, and/or immune cells (e.g., monocytes/macrophages) can be administered by any suitable means, such as by bolus infusion, by injection, such as 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).

In some embodiments, proinflammatory agents, SHP-1 inhibitors, and/or immune cells (e.g., monocytes/macrophages) are administered parenterally, intrapulmonary, and intranasally, and (where necessary for local treatment) intralesional or intratumorally. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. In some embodiments, proinflammatory agents and/or SHP-1 inhibitors are administered orally.

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

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 days. In some embodiments, the SHP-1 inhibitor and the proinflammatory agent are administered within about 24, 16, 12, 8, 4, 2, or 1 hours. In some embodiments, the SHP-1 inhibitor and the proinflammatory agent are administered within 30 minutes.

It is also contemplated that the SHP-1 inhibitors and/or proinflammatory agents described herein may be delivered via any appropriate medium or method. In some embodiments, the SHP-1 inhibitors and/or proinflammatory agents are delivered directly to tumor tissue. For this purpose, different 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, SHP-1 inhibitors and/or proinflammatory agents are delivered via nanoparticles. In some embodiments, SHP-1 inhibitors and/or proinflammatory agents are delivered via a controlled release system. In some embodiments, the SHP-1 inhibitor and/or pro-inflammatory agent is delivered via a biomaterial implantable stent. In some embodiments, the SHP-1 inhibitor and/or pro-inflammatory agent is delivered via an injectable biomaterial stent. In some embodiments, the SHP-1 inhibitor and/or pro-inflammatory agent is delivered via a transdermal delivery system. See, e.g., Riley et al., Nat Rev Drug Discov. 2019 Mar; 18(3): 175-196.

In some embodiments, the SHP-1 inhibitor and/or proinflammatory agent is delivered by 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 epithelium (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 described in Nash et al.).

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

In some embodiments, a SHP-1 inhibitor and/or a proinflammatory agent is delivered via a method or device that promotes delivery to a specific organ (e.g., an organ with a tumor). See, e.g., 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 entirety) for examples of such methods or devices.

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. VI. Compositions comprising SHP-1 inhibitors

The present application also provides a composition (eg, a pharmaceutical composition) comprising a SHP-1 inhibitor, a pro-inflammatory agent and/or an immune cell for use in treatment as described above.

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

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

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.

In some embodiments, a composition (e.g., a pharmaceutical composition) is provided, which 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 immune cells (such as monocytes or macrophages described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier.

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

In some embodiments, a composition (e.g., a pharmaceutical composition) is provided, which comprises 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 (such as a monocyte or macrophage as described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier.

In some embodiments, a composition (e.g., a pharmaceutical composition) is provided, which comprises 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 (such as a monocyte or macrophage as described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier.

In some embodiments, a composition (e.g., a pharmaceutical composition) is provided, which comprises 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 (such as monocytes or macrophages described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier.

In some embodiments, a composition (e.g., a pharmaceutical composition) is provided, which comprises a SHP-1 inhibitor (e.g., TPI-1 or an analog or derivative thereof) and an agent for use in radiation therapy (such as any radiation therapy described herein). In some embodiments, the composition further comprises an immune cell (such as a monocyte or macrophage described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier.

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

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

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 (such as any oncolytic virus described herein). In some embodiments, the composition further comprises an immune cell (such as a monocyte or macrophage described herein). In some embodiments, the composition further comprises a pharmaceutically acceptable carrier.

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

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

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

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

The following examples are intended to be merely illustrative of the present invention and therefore should not be considered to limit the present invention in any way. The following examples and detailed descriptions are provided in an illustrative and non-limiting manner. Example 1.

SHP-1 is abundantly expressed in macrophages. Proteomic studies examining the expression of non-receptor protein tyrosine phosphatases (PTPs) in macrophages showed that SHP-1 had the highest expression level. See 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 inhibitory receptor (iR) for macrophages involved in the activation of 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 phenotypic activation. (The housekeeping gene GAPDH was also detected.)

Despite its high expression, we found in our analysis of tumor-associated macrophages that SHP-1 activity in macrophages was low unless the macrophages were “surrounded/contacted” by tumor cells and simultaneously stimulated with TLR agonists or other proinflammatory activators. See Figure 2A. Under these conditions, inhibition of SHP-1 activity by the covalent inhibitor TPI-1 (8) reduced SHP-1-mediated protein dephosphorylation, resulting in a significant enhancement of signal transduction mediated by TLRs and other factors.

As shown (Fig. 2C), macrophages stimulated with IFNγ/LPS completely inhibited phosphorylation and thus activated STAT-1 (pSTAT-1) and Erk1/2 (pErk1/2) in the presence of cancer cell junctions, while inhibition of SHP-1 exerted by cancer cells released dose-dependently by TPI-1 allowed signal transduction and activation of STAT-1 (pSTAT-1) and Erk1/2 (pErk1/2). Consistently, this effect of TPI-1 inhibition of SHP-1 led to elevated proinflammatory cytokine production by macrophages and the manifestation of immunogenic antigen presentation mechanisms (Fig. 2D and Fig. 2E). Macrophages with high SHP-1 activity in the tumor environment have reduced pro-inflammatory interleukin production but high IL-10 production 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, independent of cancer cells expressing CD47 (Figures 2F and 2G). Example 2.

In addition to TPI-1, several other SHP-1 inhibitors were tested, such as PTP inhibitor-I, PTP inhibitor-III, and vitamin E (9, 10) and pseudopteroyloxanthrone dimers A and B (PXA and PXB) (11) reported in recent years. Given that PKCθ regulates SHP-1 activity via phosphorylation at Ser591 (12, 13), PKCθ inhibitors and activators were also tested in various assays.

As shown in Figures 3A to 3E, among these compounds, TPI-1 showed the strongest effect and strongly inhibited SHP-1 activity at lower concentrations. Vitamin E derivatives and PKCθ activator PMA moderately inhibited SHP-1. We also studied the effect of inhibition of SHP-2, a close family member of SHP-1. Compared with the inhibition of SHP-1, the inhibition of SHP-2 did not greatly reduce the PTP activity induced by proinflammatory activation and cancer cell binding, nor did it enable activated macrophages to phagocytose cancer cells. These studies infer that SHP-2 is regulated differently from SHP-1, and SHP-1, but not SHP-2, controls macrophage proinflammatory responses, immunogenic antigen presentation, and phagocytosis toward cancer cells. Example 3.

SHP-1 was found to have a rapid turnover rate. Treatment of macrophages with the covalent inhibitor TPI-1 for 15 min followed by washing and removal of inhibitor availability revealed robust SHP-1 inhibition immediately following TPI-1 treatment, an effect associated with increases in pSTAT-1 and pErk1/2 by IFNγ/LPS. However, this pattern of pulsatile SHP-1 inhibition did not persist beyond a few hours (5-8 h) when SHP-1 activity began to recover and reached more than 50% in 12-16 h, acquiring the ability to reduce IFNγ/LPS-induced signaling (Fig. 4A). Despite changes in SHP-1 activity, total SHP-1 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). Example 4.

Further molecular and cellular signaling studies demonstrated that in a tumor environment, tumor cells ligate macrophage iRs (e.g., SIRPα, LIIRB, and Siglec) via their cell surface counterreceptors (e.g., CD47, MHC, and carbohydrates), which in turn drive high SHP-1 activation via their cytoplasmic ITIMs that undergo tyrosine phosphorylation in the presence of proinflammatory stimuli, and bind to SHP-1, thereby disengaging the SH2 domain from SHP-1 autoinhibition (depicted in FIG5A ). Neither iR extracellular ligation nor proinflammatory stimuli alone drive robust ITIM phosphorylation and, therefore, SHP-1 activation, but their simultaneous presence achieves this drive. The underlying mechanism suggests that extracellular iR induces its cytoplasmic structural change to an "open" form, exposing ITIM to phosphorylation, and proinflammatory signals simultaneously induce activation of Src family tyrosine kinases (TKs) to mediate ITIM phosphorylation. Figure 5B shows the study of SIRPα, an essential macrophage iR, whose ITIM phosphorylation requires both extracellular CD47 and macrophage stimulation with cytokines or TLR agonists.

These studies also found that macrophage iRs (such as SIRPα) can recruit both SHP-1 and SHP-2 via cytoplasmic ITIM phosphorylation. However, when macrophages were stimulated with activating interleukins (TNFα, IL-17A, IL-6, or IFNγ) or TLR agonists (LPS, CpG, or poly-IC), SHP-1 binding occurred only under proinflammatory conditions; while SHP-2 binding was dominated by immunosuppressive IL-4, IL-10, or TGFβ (Figure 5C). Example 5

Similar findings were obtained in real tumors in vivo. As tumors grew to larger sizes, intratumoral macrophages were found to increase the expression of iRs, while tumor cells in the same TME also elevated their counterreceptors (such as CD47) and the T cell inhibitory molecule PD-L1 (Figure 6A). These changes in advanced tumors indicate that stronger immunosuppression has been established compared to their early stages. In the absence of therapy, intratumoral macrophages that are constantly connected by surrounding tumor cells are guided by TME immunosuppressive signals, resulting in phosphorylation of their iRs in cytoplasmic ITIMs and binding to SHP-2 rather than SHP-1 (data not shown). Our accumulating data indicate that iR-SHP-2 binding in this manner sequesters SHP-2 from access to immunosuppressive interleukin receptors (e.g., IL-4R and IL-10R) and thus prevents SHP-2 from inhibiting anti-inflammatory signaling (14). Thus, this iR-SHP-2 binding in established solid tumors in vivo serves as a feed-back regulator that enhances the anti-inflammatory phenotype of macrophages, thereby promoting TME immunosuppression and tumor progression.

SHP-1 activity was low in non-treated solid tumors (see Figures 7A to 7D), and this low activity inhibited 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 those of others (14-17)). Consistently, inhibition of SHP-1 in MC38 solid tumors led to increased IL-10 production in the TME (Figure 6B). Interestingly, inhibition of SHP-1 also increased tumor production of IL-6, an interleukin that has been reported to play an immunosuppressive role in the TME and support tumor progression (18, 19). In vitro analysis of macrophage activation in a tumor-related setting with 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 interleukin production.

Supporting this notion, we found that once tumors were treated therapeutically, such as with TLR ligands (αTLR), inflammatory interleukins (IL-1/6/TNFα/IFNγ), STING activators (2'3'-cGAMP), RT, anti-PD-L1 immune checkpoint blockers (αPD-L1), or the chemotherapeutic drug azacitidine (AZA), they induced a spike in SHP-1 activity (Figure 7A and 7B). In fact, macrophages with high iR expression in advanced and larger tumors provided the ability to robustly activate SHP-1 under therapeutic treatment. We found that intratumoral macrophages are the major source of SHP-1 activity, and that depletion of macrophages in tumors after therapy largely reduced SHP-1 activity spikes (Figure 7C). In addition to inactivating proinflammatory signaling pathways, this high SHP-1 activity also enhances proinflammatory stimulus-induced IL-10 and TGFβ (Figure 7D, see also Figure 2D) through currently unidentified mechanisms, thereby accelerating the recovery of TME from immunosuppression. 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 toward anti-tumor immunogenicity, such as the reprogramming we observed in pancreatic ductal adenocarcinoma (KPC) and colorectal cancer (MC38) under treatment (Figure 7D to 7F). Instead, these tumors arbitrated strong resistance to therapy through increased TGFβ and TGFβ receptors (TGFBR1 and TGFBR2) and CCL2 chemokines that attract MDSCs (Figure 7F), resulting in wound healing and enhanced immunosuppression. In cases of robust treatment resistance, there is no or only marginal antigen presentation in the TME, and anti-tumor T cell immunity is suppressed.

In stark 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 (6-18 h) after treatment (TPI-1 + αTLR or RT), intratumoral macrophages changed their phenotype from immunosuppressive to characteristically proinflammatory, characterized by high expression of proinflammatory interleukins TNFα, IFNα/β, IFNγ, IL-1β, IL-6, IL-12, IL-17, IL-18, etc., while also reducing IL-10 and TGFβ. A significant immunogenic antigen presentation mechanism was induced, with an increase in cell surface markers MHC-I, MHC-II, and co-stimulatory molecules CD80, CD86, CD40, OX40L, etc. A group of interleukins that attract neutrophils, NK cells, and T cells in the TME also increased, while TGFR and CCL2 decreased (Figure 7D to Figure 7F). Example 6

Consistent with these changes, TME analysis of MC38 colorectal tumors and KPC pancreatic tumors treated with TLR agonists in combination with TPI-1 or RT (Figures 8A to 8E and 9) revealed rapid infiltration of inflammatory neutrophils (ROShigh) and tumor-killing NK cells (Granzhigh), and strong antigen presentation also caused an expansion of tumor-specific (p15E-reactive) cytotoxic T cells (Granzhigh) with high tumor-destructive capacity. The immunosuppressive compartments (including MDSCs and Tregs) in the same TME were reduced. Interestingly, the intratumoral macrophage (F4/80+) population was also reduced to a micro-persistent after T cell activation. We further found that this T cell activation and expansion event was driven primarily by in situ intratumoral APC-mediated antigen presentation of tumor-specific memory T cells (TEM/CM) in activated TILs. Infusion of TLR agonists and TPI-1 together ex vivo, rather than separately, into resected tumors induced a similar T cell expansion response. However, treatment of resected tumors did not result in an increase in neutrophils and NK cells, indicating that the increased infiltration of these killer cells in tumors in vivo is via tropistic recruitment outside the tumor. Together, these results suggest that inhibiting SHP-1 to remove a central mechanism across pro-inflammatory signaling therapeutically unleashes anti-tumor potential within tumors, thereby empowering both innate and adaptive immune cells against cancer.

Furthermore, these analyses of changes in the immune landscape of the TME following TPI-1 combination therapy informed the design of a pulsed - intermittent SHP-1 inhibition strategy for metastatic solid tumors. We found that in tumors treated with TPI-1 in combination with a TLR agonist or RT, the intratumoral macrophage population in the TME was greatly reduced following antigen presentation to activate T cells. In addition to the macrophage reduction, tumor-associated SHP-1 activity was also reduced, a change that eliminated the necessity for continuous TPI-1 application. This "interval" period lasted 3-6 days, with the length depending on the TPI-1 and TLR agonist/RT doses and the tumor type and stage prior to treatment. During this time period, the TME is dominated by tumor-killing CD8 T cells, neutrophils, and NK cells, and treated tumors have limited growth but present a state of stable disease (SD) or tumor regression. However, assuming that the tumor is not completely eliminated, the TME is later repopulated with "new" macrophages and shows an increase in MDSCs and Tregs, indicating reestablishment of immune suppression. Consistent with these changes, tumors resume growth after the interval unless another cycle of TPI-1 combination therapy is applied, which again effectively suppresses the tumor. Example 7

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 provides a theoretical basis, but iSHP-1 in combination with pro-inflammatory regimens predicts potent tumor inhibition. Given that intratumoral macrophages indicate TME responses 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 intratumoral macrophage population dynamics, iSHP-1 treatment was given in a " pulse " when macrophages were abundant in the TME, but stopped when they were decreasing following antigen presentation to activate T cells (" rest " period). In addition, given our finding that macrophages repopulate the TME after the rest period, multiple treatment cycles were designed with each cycle repeating the pulse of iSHP-1 immediately after the rest period, assuming that the tumor was not eliminated.

Figures 10A-10B depict 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 (1× per day) at the beginning of each cycle, and iSHP-1 treatment was stopped during the 2-9 day interval between cycles. 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 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 iSHP- ¹ treatment was initiated when the size of a single tumor reached ≥ 200 ^mm3 or the total tumor burden was ≥ 300 mm3. These models included pancreatic cancer (KPC and Pan02), colorectal cancer (MC38), metastatic breast cancer 4T1, lung cancer (LLC), and T-cell lymphoma (EL4). All treatments to mice were given systemically, either by intraperitoneal (ip) or subcutaneous (sc) administration, in order to achieve a systemic effect on "metastatic" lesions. In a subset of experiments, treatments were also given by intratumoral injection (it).

Antitumor efficacy (tumor volume changes and survival rate), side effects (weight loss, proteinuria, anemia, and clinical discomfort), macro-organ toxicity and tissue inflammation due to toxicity, and changes in TME immune status were analyzed at different time points and at the end point throughout the treatment.

As will be seen in the next few examples, we have intensively reviewed the pulse - intermittent iSHP-1 strategy in preclinical tumor models, and in all cases, pulse 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. Application of this strategy by a systemic approach resulted in control and regression of cancer lesions throughout the body, resulting in higher survival rates and durable anti-tumor immunity.

More importantly, the pulsed - intermittent iSHP-1 strategy provides for the first time a practical approach that allows SHP-1 inhibition to potentially be applied as an in vivo treatment regimen, as this strategy minimizes the toxicity caused by SHP-1 deficiency. In contrast to animals deficient in SHP-1 or treated continuously with a SHP-1 inhibitor, which developed marked lung inflammation, kidney damage, colitis, anemia, and splenomegaly, mice treated with three cycles of pulsatile - intermittent iSHP-1, as well as TPI-1 in combination with TLR ligands, STING activators, RT, αPD-1/L1, and proinflammatory cytokines at doses sufficient to induce systemic tumor elimination, did not display severe lung or kidney damage, anemia, or splenomegaly (data are shown in the next 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 which eliminates prolonged depletion of SHP-1 activity in essential organs (e.g., lung, intestine, kidney, and spleen), where SHP-1 activity is critical for preventing unwanted autoimmune inflammatory responses to commensal microorganisms or detrital particles. Our further studies support this view, demonstrating that after pulsed inhibition of SHP-1 in macrophages with the covalent inhibitor TPI-1, macrophages are able to restore SHP-1 activity within 24 h. Example 8

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 cancer MC38, lung cancer LLC, and metastatic breast cancer 4T1 in fully immunocompetent murine syngeneic models. Both treatment efficacy and treatment safety were evaluated according to Figure 10.

Murine models (single or multifocal solid tumors): 1) pancreatic ductal adenocarcinoma (KPC or KPC-luc)-C57BL6 syngeneic implants, 2) colorectal cancer (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.

Tumor Model Establishment: Healthy cultured cancer cells (1-5 × 10 ⁵ ) suspended in 50 μL PBS were injected subcutaneously (sc), intraperitoneally (ip), intravenously (iv), or orthotopically into WT C57BL6 or BalbC mice (6-8wk, male and female) to establish a syngeneic model. For multiple tumor lesions (metastatic model), tumor cells were implanted in multiple locations via multiple sc injections combined with ip or iv. Palpable subcutaneous or orthotopic (e.g., 4T1) tumors were formed after 10-14 days. Tumor length and width were measured using a caliper, and tumor volume (V) was then calculated using the formula: Volume = (length × ^width2 )/2. For tumor cells expressing luciferase (KPC-luc, LLC-luc, 4T1-luc), whole-body images of luminescence intensity were obtained to visualize the tumor.

Therapeutic treatment: Treatment was initiated when single tumors grew to ≥ 200 ^mm3 or total tumor burden was ≥ 300 ^mm3 . The covalent SHP-1 inhibitor TPI-1 was selected for its potent inhibitory capacity and relative specificity. Targeting systemic effects at locations distant from 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, it). TPI-1 was administered prior to or with the combination modality. A. Study-1 (Figures 11A to 11E): Continuous and intermittent iSHP-1 in combination with TLR agonists for treatment efficacy and adverse toxicities

Tumor Model: Single Implanted KPC Pancreatic Cancer

Treatment and dosing strategy: i) iSHP-1 - TPI-1, 1, 3 and 10 mg/kg; i.p., 1× daily (continuous), or intermittently as described in Figure 11. ii) TLR agonists (αTLR)-CpG, poly I:C, 10 μg each; i.p. every 3 days

Results: TPI-1 exhibited dose-dependent effects, 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 (1× daily) caused acute anemia, proteinuria, splenomegaly, and lung inflammation. In mice treated intermittently with TPI-1, these side effects were absent or mild. In conclusion, the intermittent iSHP-1 strategy greatly reduced the risk of adverse toxicity while achieving tumor inhibitory efficacy. B. Study-2 (Figure 12A to Figure 12D): Pulse-intermittent iSHP-1 combined with TLR agonists and/or ICB (αPD-L1) in the treatment of multifocal colorectal cancer, a study targeting efficacy and adverse toxicity

Conclusions: i) Intermittent iSHP-1 in combination with TLR agonists (αTLR), or αTLR and αPD-L1 effectively regressed multiple lesions in MC38 colorectal cancer, which were otherwise resistant to TLR or αPD-L1 alone. ii) TPI-1 administered via i.p. and s.c. 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 toxicity. C. Study-3 (Figure 13A to Figure 13E): Pulse-intermittent iSHP-1 in combination with RT and αPD-L1 in multifocal pancreatic ductal adenocarcinoma and lung cancer

Modality and dosing strategy: i) iSHP-1- TPI-1, 3 or 5 mg/kg, i.p., 1× every 3 days. ii) Tumor lesion RT: 8Gy-4Gy-2Gy, given simultaneously with iSHP-1 into the right flank. iii) αPD-L1: 100 μg, i.p., given one day after iSHP-1 plus RT.

Results: Systemic intermittent iSHP-1 combined with right flank tumor-lesion RT, followed by αPD-L1 for enhanced T cell immunity, induced potent anti-tumor immunity and distal effects, thereby effectively eliminating or inhibiting KPC pancreatic cancer and LLC lung cancer as well as distal lesions. D. Study-4 (Figures 14A to 14E): Pulse-intermittent iSHP-1 combined with TLR agonists for advanced, larger KPC pancreatic cancer.

Mouse model: Single large pancreatic duct adenocarcinoma (KPC-luc) C57BL6 syngeneic type

Treatment: Combination of SHP-1 inhibitor (iSHP-1) and TLR agonist (αTLR)

Results: As shown in Figures 14A to 14E, TPI-1 combined with αTLR enhanced anti-tumor immune cells (such as CD8+ T cells, NK cells, and neutrophils) and effectively eliminated advanced, larger KPC pancreatic cancers. Example 9

In vitro macrophages were analyzed in the presence of macrophage iR-linked tumor cells with or without TPI-1 inhibition of SHP-1 (iShp1). See the test system in Figure 16A. As shown in Figures 16B and 16C, IFNα treatment alone did not induce antigen presentation or proinflammatory responses by macrophages without and with iShp1. IFNγ plus iShp1 significantly increased antigen presentation, even though the combination did not induce proinflammatory interleukin production. In contrast, IL-1 family interleukins (IL-1β, IL-18), TNFα, and TLR ligands in combination with iShp1 demonstrated the ability to proinflammatoryally activate macrophages in a tumor environment. Example 10

MC38 colorectal cancer was established (by subcutaneous administration) in syngeneic C57BL6 mice. After tumor size reached over 200 mm ³ , systemic inflammatory symptoms were induced in tumor-bearing mice by TLR agonists (αTLR, CpG/poly IC/R848, 25 μg each) administered by subcutaneous administration at a site distal to the MC38 tumor. One group of mice was concurrently administered TPI-1 (1 mg/kg) by subcutaneous administration to achieve systemic SHP-1 inhibition.

As shown in Figure 17A, MC38 tumors were protected from αTLR-induced acute inflammation by SHP-1. 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 unlocked tumor immunosuppression and enabled neutrophil infiltration after αTLR (αTLR + TPI-1).

As shown in FIG. 17B , TEM analysis confirmed increased neutrophil infiltration in tumor tissues in mice treated with αTLR plus TPI-1.

As shown in Figure 17C, inhibition of SHP-1 by TPI-1 enables intratumoral macrophages to skew toward pro-inflammatory activation via αTLR, thereby displaying increased TNFα and IL12 expression. In contrast, the absence of TPI-1 causes macrophages to resist pro-inflammatory activation via αTLR, while enhancing immunosuppression and increasing IL-10 and TGFβ expression. Example 11

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

Mice with established MC38 colorectal cancer (200-400 mm ³ ) 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 tumor TME was analyzed on day 6 after treatment. See Figure 11A.

As shown in Figure 11B, tumor volume was reduced after αTLR + TPI-1 + dasatinib treatment, and the administration of anti-TNFα or anti-IL-6 did not interfere with its anti-tumor 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 induced by αTLR/TPI-1/dasatinib therapy and the decrease in macrophages and MDSC in the TME. See Figure 11C and Figure 11D. Treatment of mice with anti-TNFα mAb but not anti-IL-6 mAb greatly reduced the induction of inflammatory cytokines (TNFα, IL-6, IL-1β, IL-10, IFNα and IFNγ) associated with αTLR/TPI-1/dasatinib combination therapy. Anti-TNFα treatment also significantly reduced circulating monocyte and PMN interleukins CCL2, CCL5, and CXCL1, while not reducing CXCL10, which is essential for T cell trafficking. FIG. 11E . Furthermore, as shown in FIG. 11F , anti-TNFα treatment protected mice from developing splenomegaly and intestinal inflammation typically associated with αTLR/TPI-1/dasatinib therapy.

In summary, the results showed that neither anti-TNFα nor anti-IL-6 interfered with TKi/iShp1/αTLR-driven anti-tumor immunity or achieved therapeutic efficacy. In addition, anti-TNFα exhibited beneficial effects by largely eliminating TKi/iShp1/αTLR-induced interleukin storm and systemic inflammation, thereby inhibiting therapy-related adverse toxicities.

In addition, although the above specific experiments involved the use of both TPI-1 and dasatinib and both poly I:C and R848, our results from experiments using either TPI-1 and dasatinib and either poly I:C and R848 achieved similar effects (data not shown). We also found that the appropriate time window for anti-TNFα antibody treatment can be at least a week before (as long as the antibody is stable within the time window) to immediately after (e.g., within 0.5-1 hour) SHP-1 inhibitor/αTLR treatment. Preferably, the anti-TNFα antibody is provided before or simultaneously with the SHP-1 inhibitor and/or αTLR so that it maximally blocks TNFα induced after SHP-1 inhibitor and proinflammatory agent treatment. Example 12

To investigate the mechanisms by which tumor cells inhibit pro-inflammatory responses of macrophages in the TME. As shown in Figure 19, Panel A, macrophage responses in the tumor environment were analyzed: human monocyte-derived macrophages (MØ) were cultured alone (configuration 1), in the presence of cancer cell-conditioned medium (50%) containing cancer cell-secreted factors (secretomes; configuration 2), or co-cultured with cancer cells (cell "contact" model; configuration 3), or co-cultured with cancer cells placed in transwells (0.4 µm) without "contact" of MØ (configuration 4). 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).

Next, the macrophage response to R848/IFNγ was studied. The presence of cancer secretomes (configurations 2) and 4)) did not alter the pattern of macrophage proinflammatory response, although a partial inhibition of proinflammatory interleukin production (10%-40% reduction) was produced. In contrast, macrophages co-cultured with cancer cells abolished the proinflammatory response and exhibited enhanced immunosuppression and high IL-10 production. See Figure 19, Panel B.

Macrophage responses to R848 or MSA-2 were also studied. Results similar to those in Figure B: The presence of cancer secretomes partially inhibited macrophage proinflammatory responses, but macrophages co-cultured with cancer cells showed abrogated proinflammatory responses and enhanced immunosuppression with high IL-10 and TGFβ production. See Figure 19, Figure C.

Furthermore, as shown in FIG20 , macrophages co-cultured with cancer cells abolished TLR agonist (R848) plus IFNγ-induced macrophage antigen presentation. Example 13

It has been found that when the tumor progresses to an advanced stage (larger size), iR and its ligands are upregulated, as shown in Figure 21. Panel A depicts the immune cell composition and percentages in a typical MC38 colorectal cancer. Panel B depicts the expression of multiple inhibitory receptors (iR) on myeloid immune populations (including TAM (F4/80+), MDSC (Ly6C+), and N2-neutrophils (PMN)) in MC38 tumors of different sizes. Panels C to D depict the expression of CD47 (receptor for SIRPα) and PD-L1 on the same MC38 tumors of different sizes as in Panel B. Panel E depicts representative IHC staining of samples of human cancers showing increased expression of ligands for myeloid iR. Example 14

It was also found that factors (secreted proteins) produced by cancer cells and tumor TME induce increased macrophage expression of iR. See, for example, Figure 22. Panel 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 Panel B of Figure 22.

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, kidney cancer TK10, ovarian cancer OVCAR3 and monocytic leukemia THP1. Examples of iR tested on human macrophages are: SIRPα, LILRB1, LILRB4 and Siglec7. See Figure 22, Panel C. Interleukins produced by mouse and human cancer cells in culture are shown in Figure 22, Panel D. Treatment of macrophages with cancer cell interleukins induces an increase in iR expression (SIRPα as an example) on macrophages (bone marrow-derived macrophages (BMDM) and peritoneal macrophages (PEM)). See Figure 22, Panel E. Example 15

Inhibition of SHP-1 unleashes pro-inflammatory responses in the KPC tumor TME, as shown in Figures 23A to 23B. KPC pancreatic tumors explanted from mice were cut into small pieces and treated with 5 different TLR agonists (R848, 3M-852A, motomod, bropirimide, or visammod; 1 μg/ml each) without or with the SHP-1 inhibitor TPI-1 (0.4 μM). After 18 h, interleukins secreted in the culture medium were analyzed.

As shown in Figure 23A, all five TLR agonists failed to trigger proinflammatory interleukin production but induced high levels of IL-10 and TGFβ, indicating enhanced immunosuppression in the TME.

As shown in Figure 23B, inhibition of SHP-1 (iSHP-1) enables TLR agonists to drive pro-inflammatory responses in tumor tissues. As shown, the addition of the SHP-1 inhibitor TPI-1 to A) induced high pro-inflammatory interleukin production while inhibiting the production of IL-10 and TGFβ. Example 16

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 μg/ml each) ± TPI-1 (0.4 µM). After 18 h, interleukins secreted in the culture medium were analyzed. As shown in Figure 24, αTLR plus TPI-1 induced a high pro-inflammatory response while suppressing IL-10 in the TME. Example 17

The iR → SHP-1 inhibitory axis was investigated. SHP-1 activated by iR dephosphorylates inflammatory stimulus-induced JAK-STAT, NFĸB, MAPK, and PI3K-Akt activation pathways, thereby quenching pro-inflammatory signaling and conferring therapeutic resistance. Inhibition of SHP-1 abolishes multiple axis iR-mediated inhibitory regulation, thereby releasing macrophage pro-inflammatory polarization, as shown in Figures 25A-25B. Example 18

As shown in Figures 26B and 26C, under TLR and IFNγ stimulation, Shp1 ^-/- macrophages resist the inhibition imposed by cancer cells and release proinflammatory responses. In the presence of B16 melanoma cells, murine bone marrow-derived macrophages (BMDM) prepared from WT or isotype-linked Shp1 ^-/- mice were treated with TLR agonists (αTLR; R848, 1 µg/ml) plus IFNγ (40 ng/ml). See Figure 26A. After 16 h, the cell culture medium was collected and analyzed for cytokines. The results are shown in Figure 26B. Macrophages were also collected and analyzed for cell surface expression of their inflammatory phenotype and antigen presentation mechanisms. The results are shown in Figure 26C. Example 19

As shown in Figures 27A-27C, cell surface blockade of iR or ligand is an alternative strategy to deplete the inhibitory iR→SHP-1 axis.

Experimental setup: Human macrophages co-cultured with cancer cells were stimulated with pro-inflammatory 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 mAb for Siglec (αSiglec-7, -8, or -9), mAb that blocks LilRB-MHC interaction (αLILRB1, αLilRB2, αLIlRB3, αLilRB4, or pan-HLA-A/B/C blocking Ab), or mAb that blocks CD47-SIRPα interaction (αCD47 or αSIRPα). To remove sialic acid structures on the cancer cell surface that bind to Siglec on macrophages, cancer cells were also treated with neuraminidase (50 mU/ml, 1 h) before use in the experiment. See Figure 27A.

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

Examples of data (macrophages and SW260 cancer cells). As shown in Figure 27C, blockade of a single iR-ligand axis is not sufficient to remove the immunosuppression imposed by cancer cells on macrophages, while blocking multiple iR-ligand interactions or inhibition of SHP-1 downstream of all iRs in macrophages eliminates tumor cell suppression, thereby releasing macrophages for proinflammatory responses. Similar results were obtained when macrophages were co-cultured with other cancer cells (such as OVCAR3, MDA231, TK10, HT29, etc.). References: 1. Kang, XL, J. Kim, M. Deng, S. John, HY Chen, GJ Wu, H. Phan, and CC Zhang. 2016. Inhibitory leukocyteimmunoglobulin-like receptors: Immune checkpoint proteins and tumor sustaining factors. Cell Cycle 15: 25-40. 2. Zarrin, AA, and RC Monteiro. 2020. Editorial: The Role of Inhibitory Receptors in Inflammation andCancer. Front Immunol 11. 3. Carosella, ED, S. Gregori, and D. Tronik-Le Roux. 2021. HLA-G/LILRBs: A Cancer Immunotherapy Challenge. Trends Cancer 7: 389-392. 4. Yanagita, T., Y. Murata, D. Tanaka, SI Motegi, E. Arai, EW Daniwijaya, D. Hazama, K. Washio, Y. Saito, T.Kotani, H. Ohnishi, PA Oldenborg, NV Garcia, M. Miyasaka, O. Ishikawa, Y. Kanai, T. Komori, and T.Matozaki. 2017. Anti-SIRP alpha antibodies as a potential new tool for cancer immunotherapy. Jci Insight 2. 5. Zhang, W., Q. Huang, W. Xiao, Y. Zhao, J. Pi, H. Xu, H. Zhao, J. Xu, CE Evans, and H. Jin. 2020. Advances inAnti-Tumor Treatments Targeting the CD47/SIRPαlpha Axis. Front Immunol 11: 18. 6. Green, MC, and LD Shultz. 1975. Motheaten, an immunodeficient mutant of the mouse. I. Genetics andpathology. J Hered 66: 250-258. 7. Shultz, LD, and MC Green. 1976. Motheaten, an immunodeficient mutant of the mouse. II. Depressedimmune competence and elevated serum immunoglobulins. J Immunol 116: 936-943. 8. Kundu, S., K. Fan, ML Cao, DJ Lindner, ZZJ Zhao, E. Borden, and TL Yi. 2010. Novel SHP-1 InhibitorsTyrosine Phosphatase Inhibitor-1 and Analogs with Preclinical Anti-Tumor Activities as Tolerated Oral Agents. J Immunol 184: 6529-6536. 9. Yuan, X., Y. Duan, Y. Xiao, K. Sun, Y. Qi, Y. Zhang, Z. Ahmed, D. Moiani, J. Yao, H. Li, L. Zhang, AE Yuzhalin, P.Li, C. Zhang, A. Badu-Nkansah, Y. Saito, X. Liu, WL Kuo, H. Ying, SC Sun, JC Chang, JA Tainer, and D. Yu.2022. Vitamin E Enhances Cancer Immunotherapy by Reinvigorating Dendritic Cells via Targeting CheckpointSHP-1. Cancer Discov . 10. Arabaci, G., XC Guo, KD Beebe, KM Coggeshall, and D. Pei. 1999. alpha-Haloacetophenone derivatives as photoreversible covalent inhibitors of protein tyrosine phosphatases. J Am Chem Soc 121: 5085-5086. 11. Yang, RL, Q. Dong, HB Xu, XH Gao, ZY Zhao, JC Qin, C. Chen, and DQ Luo. 2020. Identification ofPhomoxanthone A and B as Protein Tyrosine Phosphatase Inhibitors. Acs Omega 5: 25927-25935. 12. Ben-Shmuel, A., B. Sabag, A. Puthenveetil, G. Biber, M. Levy, T. Jubany, F. Awwad, RK Roy, N. Joseph, O.Matalon, J. Kivelevitz, and M. Barda-Saad. 2022. Inhibition of SHP-1 activity by PKC-theta regulates NK cellactivation threshold and cytotoxicity. Elife 11. 13. Jones, ML, JD Craik, JM Gibbins, and AW Poole. 2004. Regulation of SHP-1 tyrosine phosphatase inhuman platelets by serine phosphorylation at its C terminus. J Biol Chem 279: 40475-40483. 14. Shi, L., K. Kidder, Z. Bian, SKT Chiang, C. Ouellette, and Y. Liu. 2021. SIRPαlpha sequesters SHP-2 topromote IL-4 and IL-13 signaling and the alternative activation of macrophages. Sci Signal 14: eabb3966. 15. Huang, Z., JM Coleman, Y. Su, M. Mann, J. Ryan, LD Shultz, and H. Huang. 2005. SHP-1 regulates STAT6phosphorylation and IL-4-mediated function in a cell type-specific manner. Cytokine 29: 118-124. 16. Johnson, DJ, LI Pao, S. Dhanji, K. Murakami, PS Ohashi, and BG Neel. 2013. SHP-1 regulates T cellhomeostasis by limiting IL-4 signals. J Exp Med 210: 1419-1431. 17. Tao, B., W. Jin, J. Xu, Z. Liang, J. Yao, Y. Zhang, K. Wang, H. Cheng, X. Zhang, and Y. Ke. 2014. Myeloid-specific disruption of tyrosine phosphatase Shp2 promotes alternative activation of macrophages and predisposesmice to pulmonary fibrosis. J Immunol 193: 2801-2811. 18. Fisher, DT, MM Appenheimer, and SS Evans. 2014. The two faces of IL-6 in the tumormicroenvironment. Semin Immunol 26: 38-47. 19. Chonov, DC, MMK Ignatova, JR Ananiev, and MV Gulubova. 2019. IL-6 Activities in the TumourMicroenvironment. Part 1. Open Access Maced J Med Sci 7: 2391-2398.

Figure 1 depicts that SHP-1 functions as a “master” signaling mediator downstream of multiple inhibitory receptors on bone marrow 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 immunosuppressive phenotype of bone marrow leukocytes. Our approach to SHP-1 inhibition as an anti-cancer strategy (red). Some of the companies and approaches that aim to deplete or block inhibitory receptors on the surface of individual cells include SIRPα (SIRPαnt Immunotherapeutics, anti-CD47 Gilead) and anti-SIRPα (Biosion) approaches), Siglec (NextCure), LilRB (Next-IO), SLAMF (BMS), etc.

Figures 2A to 2G depict that inhibition of SHP-1 enhances macrophage proinflammatory responses, antigen presentation, and phagocytosis in the tumor environment. Figures 2A and 2B depict that SHP-1 activity in macrophages is triggered by proinflammatory stimuli and extracellular cancer cells binding ("contacting") macrophages (green bars in Figure 2A). SHP-1 activity was inhibited in a dose-dependent manner by the catalytic domain covalent inhibitor TPI-1 (1 µg/ml, Figure 2A) (Figure 2B). The SHP-2 inhibitor SHP099 (1 µg/ml) was used in the analysis. Figure 2C depicts 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 line). Figures 2D and 2E depict that TPI-1 inhibition of SHP-1 in macrophages enhances the production of pro-inflammatory interleukins (Figure 2D) and the expression of immunogenic antigen presentation mechanisms by IFNγ/LPS in the tumor environment (Figure 2E). Figures 2F and 2G depict that SHP-1 inhibition promotes pro-inflammatory activated macrophages to phagocytose cancer cells. Figure 2F shows human monocyte-derived macrophages engulfing THP-1 leukemia cells and HT29 colorectal cancer cells. Figure 2G shows murine bone marrow-derived macrophages engulfing isogenic cancer cells EL4 (T lymphoma), B16 (melanoma), MC38 (colorectal cancer), Pan01 and KPC (both pancreatic cancers), and LLC (lung cancer). The SHP-1 and SHP-2 inhibitors TPI-1 and SHP099 were used in the analysis at 1 µg/ml, respectively.

Figures 3A to 3E depict the regulation of SHP-1 activity in proinflammatory macrophages in cancer. Macrophages co-cultured with cancer cells were stimulated with IFNγ/LPS or TLR agonists (αTLR) containing CpG, polycytidylic acid (i.e., poly I:C) and R848 (0.4 µg/ml each) for 30 min (37°C) in the presence or absence of different SHP-1 or SHP-2 inhibitors or activators, followed by analysis of cell lysis and PTP activity. Figure 3A depicts the mechanism of regulation of SHP-1 activity. Figures 3B and 3C depict that PTP activity in macrophages induced by IFNγ/LPS (Figure 3B) or TLR agonists (Figure 3C) in the presence of cancer cell junctions was largely attenuated (>70%) by inhibition of SHP-1 (TPI-1 and PTP-I) or pan-PTP inhibitors PTP-III and pervanadate, but only slightly reduced (<10%) by inhibition of SHP-2 (SHP099 and PHPS1). Figure 3D depicts that vitamin E derivatives and pseudo-piperidin dimer A (PXA) dose-dependently inhibited SHP-1 activity in IFNγ/LPS-stimulated macrophages surrounding cancer cells. Figure 3E depicts that PKCθ negatively regulates SHP-1 activity to a moderate extent. Inhibition of PKCθ by PKCθ inhibitor I and VTX27 increased SHP-1 activity via IFNγ/LPS and cancer cell ligation. In contrast, activation of PKCθ by PMA decreased SHP-1 activity.

Figures 4A and 4B depict that pulse 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 min and then washed to completely remove TPI-1. At various time points after TPI-1 treatment, macrophages were stimulated with IFNγ/LPS in the presence of live cancer cells (1:1 ratio with BMDM) for 20 min, after which the cells were lysed and analyzed for SHP-1 activity using pNpp and total and phosphorylated STAT1 and Erk1/2 by WB. FIG4B : Same setup as FIG4A , except that after TPI-1 treatment, TPI-1 was partially removed (50%) or not from the culture medium, followed by macrophage stimulation with IFNγ/LPS at different time points.

Figures 5A-5C depict signaling mechanisms in solid tumors when SHP-1 is targeted. Figure 5A depicts 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 : pro-inflammatory signal → tyrosine kinase (TK) → ITIM phosphorylation → SHP-1 activation. SHP-1 activity then inhibits pro-inflammatory signaling and confers tumor resistance to therapy. TPI-1 inhibits SHP-1 activity and restores pro-inflammatory signaling, resulting in anti-tumor innate and acquired immunity. Figure 5B depicts an example of what can be seen in SIRPα. Proinflammatory signals induce one or more Src family TKs to mediate docking of SHP-1, thereby leading to phosphorylation in the cytoplasmic domain ITIM of SHP-1 activation. Figure 5C depicts that phosphorylation in iR ITIM induced by proinflammatory stimuli leads to exclusive binding of SHP-1 to pITIM, thereby leading to SHP-1 activation; while immunosuppressive signals caused by IL-4, IL-10 and TGFβ induce ITIM phosphorylation, leading to SHP-1 binding. Examples show macrophage iR LilRB and SIRPα.

Figures 6A to 6D depict that, under different conditions, inhibition of SHP-1 promotes anti-tumor or pro-tumor effects. Figure 6A depicts that intratumoral macrophages upregulate the expression of iRs (Pir-B; Siglec E, F, and G; and SIRPα) and that tumors progress to late and larger sizes. Smaller (< 150 mm ³ ) KPC pancreatic tumors and those that grew to larger sizes (> 800 mm ³ ) were dissociated into single cells and then analyzed by flow cytometry for cell surface protein expression on macrophages (gated F4/80+). Figure 6B depicts that inhibition of SHP-1 alone promotes TME immunosuppression. In cell culture, excised MC38 solid tumor cubes were treated with the SHP-1 inhibitor TPI-1 (100 nM) or vehicle (DMSO). After 24 h, tumors secreted interleukins into the culture medium were analyzed by ELISA. As shown, TPI-1 treatment increased tumor production of IL-6 and IL-10. Figure 6C depicts that TPI-1 and PTP-1 enhance macrophage production of IL-10 and TGFβ in a dose-dependent manner in the presence of cancer cell junctions under immunosuppressive stimulation by IL-4/13 or IL-10. Figure 6D with the same experimental settings demonstrated that TPI-1 and PTP-1 dose-dependently enhanced the proinflammatory response of macrophages, with an increase in IL-12 and TNFα induced by IFNγ and LPS.

Figures 7A to 7F depict that inhibition of SHP-1 unleashes proinflammatory responses and antigen presentation in solid tumors after therapy. Figure 7A shows that intratumoral treatment of KPC pancreatic tumors with TLR agonists (αTLR, which contains CpG, poly I:C, and R848, 1 µg each), proinflammatory interleukins (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 in vivo stationary tumors. FIG7B shows that, similarly, PTP/SHP-1 activity in KPC tumors was induced by a dose of 8Gy RT treatment, a dose of chemotherapy with azathioprine (AZA) or an anti-PD-L1 antibody (αPD-L1), and these activities were attenuated by simultaneous treatment with TPI-1. FIG7C depicts that depletion of intratumoral macrophages with clodronate liposomes abolished SHP-1 activity induced by multiple treatments of the tumor. FIG7D and FIG7E show that inhibition of SHP-1 greatly enhanced TLR agonists and RT-induced proinflammatory cytokines ( FIG7D ) and the ability of intratumoral macrophages to present immunogenic antigens ( FIG7E ). Of note, treatment of tumors with TLR agonists or RT in the absence of SHP-1 inhibition induced a small increase in proinflammatory interleukins, but a significant increase in IL-10 and TGFβ. Figure 7F depicts transcriptional profiles showing distinctly different tumor responses to TLR agonists and RT in the absence and with intratumoral SHP-1 inhibition. In the absence of SHP-1 inhibition, KPC tumors were resistant to treatment with increased immunosuppressive TGFβ signaling and MDSC infiltration, whereas treated tumors with SHP-1 inhibition caused TME reprogramming to a potent pro-inflammatory niche with reduced TGFβ but high expression of inflammatory cytokines, antigen-presenting molecules, and chemokines that attracted neutrophils, NK and T cells but not MDSC. Similar data were obtained by studying the colorectal carcinoma MC38.

Figures 8A to 8E depict that SHP-1 inhibition combined with TLR agonists (αTLR) reprograms the TME of MC38 colorectal cancer. Figure 8A depicts the MC38 tumor treatment regimen. Figures 8B to 8C show that TME analysis confirmed that TME was reprogrammed by combined treatment with TPI-1 and αTLR, inducing a decrease in tumor cells and an increase in immune infiltration (especially tumor-destructive CD8 T cells, neutrophils (PMNs) and NK cells, while macrophages, MDSCs and Tregs were reduced. Figure 8D shows a significant decrease in macrophages within the tumor after TPI-1 and αTLR treatment. Figure 8E shows that excised MC38 tumors treated with TPI-1 and αTLR ex vivo induced CD8 T cell expansion, indicating that the treatment induced antigen presentation in situ.

Figures 9A to 9C depict that TPI-1 inhibition of SHP-1 combined with tumor-focal RT reprograms the TME of KPC pancreatic ductal adenocarcinoma toward pro-inflammatory cancer elimination. Figure 9A depicts the treatment regimen and TME analysis at day 5. The table depicts the percentage of various populations within total CD45+ cells. The bar graph depicts the percentage of various populations within total cells. Figure 9A confirms that TPI-1 in combination with RT induces neutrophil (PMN) infiltration, NK cell increase, and CD8 T cell expansion. Figure 9B depicts that TPI-1 combined with RT treatment induces a significant expansion of CD8 T cells with a significant high frequency response to the tumor-specific antigen p15E. FIG9C depicts that intratumoral macrophages exhibited a pro-inflammatory phenotype and increased antigen presentation ability after combined treatment with TPI-1 and RT.

Figures 10A-10B depict the pulsatile intermittent SHP-1 inhibition (iShp-1) strategy for treating metastatic solid tumors. Figure 10A: A preclinical metastatic solid tumor model was established in syngeneic WT mice by multi-site 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 given ip or sc for systemic effect. Treatment could also be given by intratumoral injection (it) . FIG. 10B : Treatment regimens and assessments: Pulse intermittent regimens SHP-1 inhibitors are administered once at the beginning of each cycle, or two or three times in succession (pulse -1, -2, or -3), followed by a rest period (2-9 days variable) before the next treatment cycle. Combination modalities (not limited to the list) are administered simultaneously (e.g., TLR agonists, shown in the figure), or otherwise after a specific dosing schedule. Tumor control efficacy, side effects, and toxicity are assessed throughout the experiment. Changes in TME immunogenicity are also determined for mechanistic insights.

Figures 11A to 11E depict the effects of continuous or intermittent iShp-1 treatment on efficacy and adverse toxicity. Figure 11A depicts the study design. Mice bearing KPC pancreatic cancer were treated with TPI-1 continuously (1× daily) or in an intermittent manner (intermittent) with a gap day between two treatments. Three doses of 1, 3, and 10 mg/kg (i.p.) were tested, and a TLR agonist (CpG poly-I:C, 10 µg each, i.p., 1× every 3 days) was given to initiate an inflammatory response. Figures 11B and 11C show tumor treatment efficacy. Tumor imaging (Figure 11B) and volume change recording (Figure 11C) indicate that iShp-1 has similar efficacy in continuous or intermittent regimens. Figure 11D and Figure 11E show the side effects. Daily records of body weight, blood hemoglobin, proteinuria, and serum alanine aminotransferase (ALT) and analysis of spleen enlargement at the end point (d11) confirmed the high risk of continuous iSHP-1, which caused anemia, kidney damage, spleen enlargement, and lung inflammation (not shown). However, intermittent iShp-1 demonstrated a low risk of side effects.

Figures 12A to 12D depict that pulsatile-intermittent SHP-1 inhibition (iSHP-1) in combination with TLR agonists (αTLR) and/or anti-PD-L1 checkpoint inhibitors effectively treat multifocal MC38 colorectal cancer. Figure 12A depicts the treatment regimen. Mice with bilateral MC38 tumors were treated with TPI-1 (s.c.) and various combinations (via i.p. or s.c.) for two days. A 5-day rest period was given before the second cycle of treatment of mice with residual tumors. Figures 12B and 12C depict tumor control efficacy measured by tumor volume change (Figure 12B) and TME reprogramming, indicating an increase in tumor-killing immune populations while reducing immunosuppression. Figure 12D depicts acute adverse toxicity measured by body weight, proteinuria, serum ALT levels and spleen mass.

Figures 13A to 13E depict pulsatile-intermittent SHP-1 inhibition (iSHP-1) in combination with RT and αPD-L1 for pancreatic and lung cancer. Figure 13A depicts the experimental scheme. On day 1, mice with bilateral KPC pancreatic cancer or LLC lung cancer were treated with a single pulsatile dose of TPI-1 (3 mg/kg) via i.p. to systemically inhibit SHP-1 (iSHP-1). Concurrently, tumors in the right flank were treated with 8Gy X-ray irradiation (RT). After a two-day rest period, on day 4, mice were treated with a second cycle of TPI-1 (i.p.) at the same dose with RT reduced to 4Gy for right flank tumors. Cycle 3 (Day 7) iSHP-1 was combined with RT reduced to 2Gy. Anti-PD-L1 Ab (100 µg, i.p.) was given one day after TPI-1 plus RT. Figure 13B depicts luminescence images tracking the changes in KPC-luc and LLC-luc tumors after treatment. Figure 13C depicts the changes in tumor volume and the record of animal survival until 45 days after treatment. Figures 13D and 13E show that treatment did not cause splenomegaly, weight loss or anemia (Figure 13D), nor did it cause lung inflammation (Figure 13E).

Figures 14A to 14E depict the treatment of advanced KPC pancreatic ductal adenocarcinoma with pulsed-intermittent inhibition of SHP-1 (iSHP-1) in combination with a TLR agonist (αTLR). Figure 14A depicts the treatment regimen. Mice with larger KPC tumors were treated continuously (i.p.) for three days with TPI-I plus TLR agonists (CpG, poly IC, and R848, 50 µg each). After the initial pulse treatment, a 9-day rest period was given before the second two-day treatment with TPI plus TLR agonists. Figure 14B depicts luminescence images of mice with KPC tumors during the treatment process. Figure 14C depicts changes in tumor volume. Figure 14D depicts that TME analysis on day 4 showed a decrease in immunosuppressive populations (including macrophages (MØ) and MDSCs) within the tumor, and an increase in tumor-killing CD8 T cells (Tc), inflammatory neutrophils (PMNs), and NK cells. Figure 14E depicts that the treatment regimen caused mild side effects and temporary weight loss, which then recovered.

FIG. 15 depicts proteosome analysis of protein tyrosine phosphatase expression in macrophages.

Figures 16A-16D depict activation of macrophage proinflammatory responses and antigen presentation in the tumor environment by a combination regimen of SHP-1 inhibition (iShp1). Figure 16A depicts the test system. Figures 16B and 16C depict the effects of interferons with or without TPI-1. Figure 16D depicts the effects of various agents (including IL-1 family interleukins (IL-1β, IL-18), TNFα, and TLR ligands) in combination with TPI-1.

Figures 17A to 17C depict that inhibition of SHP-1 (iShp1) abolishes tumor-imposed immunosuppression under proinflammatory stimulation. Figure 17A depicts 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 depicts the phenotype of macrophages within tumors.

Figures 18A to 18G show that anti-TNFα mAb curbs systemic inflammation and reduces adverse toxicity. Figure 18A shows the experimental design. Mice with established MC38 colorectal cancer (200-400 mm 3 ) 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 ). The treatment was repeated once (d1 and d2). Tumor volume changes were recorded, and immune infiltration of tumor TME was analyzed on day 6 after treatment. Figure 18B shows tumor volume changes after various treatments. Figures 18C and 18D show 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 18E shows that treatment of mice with anti-TNFα mAb but not anti-IL-6 mAb greatly reduced the induction of inflammatory interleukins (TNFα, IL-6, IL-1β, IL-10, IFNα, and IFNγ) associated with αTLR/TPI-1/dasatinib combination therapy. Figure 18F shows that anti-TNFα treatment also significantly reduced circulating monocyte and PMN interleukins CCL2, CCL5, and CXCL1, while not reducing CXCL10, which is essential for T cell trafficking. Figure 18G shows that anti-TNFα treatment protected mice from developing splenomegaly and intestinal inflammation typically associated with αTLR/TPI-1/dasatinib therapy.

Figures 19A to 19C and Figure 20 show the mechanism by which tumor cells inhibit the pro-inflammatory response of macrophages in the TME.

Figures 21A to 21E show that iR and its ligands are upregulated when the tumor progresses to the late stage.

FIG. 22 shows that cancer cell- and tumor TME-produced factors (secretome) induce increased macrophage expression of iR.

Figures 23A-23B show that inhibition of SHP-1 unleashes pro-inflammatory responses in KPC tumor TME.

Figure 24 shows that treatment of MC38 tumors with a TLR agonist (αTLR) plus SHP-1 inhibition induces a pro-inflammatory polarization of the TME.

Figures 25A and 25B show the iR → SHP-1 inhibition axis. Figure 25A shows Western blot analysis of JAK-STAT, NFĸB, MAPK and PI3K-Akt signaling pathway activation and protein phosphorylation triggered by LPS (1 µg/ml) plus IFNγ (40 ng/ml) stimulation. Figure 25B shows densitometry analysis of protein phosphorylation and therefore signal transduction activation.

Figures 26A to 26C show that, under TLR and IFNγ stimulation, Shp1 ^-/- macrophages resist the inhibition imposed by cancer cells and release a pro-inflammatory response.

Figures 27A to 27C show that cell surface blockade of iR or ligand serves as an alternative strategy to deplete the inhibitory iR→SHP-1 axis.

Claims (59)

A method of treating cancer in a subject, comprising administering to the subject a) a SHP-1 inhibitor, and b) a pro-inflammatory agent, wherein the method comprises intermittently administering the SHP-1 inhibitor to the subject. A method of treating cancer in a subject, comprising administering to the subject a) a SHP-1 inhibitor, and b) a pro-inflammatory agent, wherein the method comprises administering the SHP-1 inhibitor systemically. A method for treating cancer in an individual comprises administering to the individual a) a SHP-1 inhibitor, and b) a pro-inflammatory agent, wherein the pro-inflammatory agent comprises an agent selected from the group consisting of a TLR agonist, a STING activator, a PAMP/DAMP activator, chemotherapy, a pro-inflammatory cytokine, a cancer vaccine, a bacterial component, sonic therapy, magnetic therapy, electrical therapy, and electrostatic therapy. A method of treating cancer in a subject comprises administering to the subject a SHP-1 inhibitor, wherein the subject is in an inflammatory response. The method of any one of claims 2 to 4, wherein the method further comprises intermittently administering the SHP-1 inhibitor to the individual. The method of any one of claims 1 and 3 to 5, wherein the method comprises systemically administering the SHP-1 inhibitor. 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. The method of any one of claims 1 and 5 to 7, wherein the method comprises administering the SHP-1 inhibitor to the subject for at least two cycles, wherein each cycle has about three to about twenty days. The method of any one of claims 1 to 8, wherein the SHP-1 inhibitor does not inhibit SHP-2. The method of any one of claims 1 to 9, wherein 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. The method of any one of claims 1 to 10, wherein the SHP-1 inhibitor is effective to inhibit greater than 50% of SHP-1 activity for no longer than about 5 days, optionally wherein the SHP-1 inhibitor is effective to inhibit greater than 50% of SHP-1 activity for no longer than about 3 days. The method of any one of claims 1 to 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). The method of claim 12, wherein the SHP-1 inhibitor is selected from the group consisting of TPI-1 or its analogs or derivatives, vitamin E derivatives, phomoxanthone A (PXA) and PKCθ activators. The method of claim 13, wherein the SHP-1 inhibitor comprises TPI-1. The method of any one of claims 1 to 14, wherein the SHP-1 inhibitor is administered at least three times. The method of any one of claims 1 to 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. The method of any one of claims 2 and 6 to 16, wherein the systemic administration of SHP-1 comprises oral administration, intravenous administration, subcutaneous administration and/or intraperitoneal administration. The method of any one of claims 1 to 3 and 5 to 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. The method of any one of claims 1 to 3 and 5 to 18, wherein the method comprises administering the proinflammatory agent intratumorally. The method of any one of claims 1 to 3 and 5 to 19, wherein the method comprises administering the proinflammatory agent to a site different from the site of the cancer to be treated. The method of any one of claims 1 to 2 and 5 to 18, wherein the proinflammatory agent comprises an agent selected from the group consisting of: TLR agonists, STING activators, radiation therapy, PAMP/DAMP activators, checkpoint inhibitors, proinflammatory interleukins, chemotherapeutic agents, bacterial components, cancer vaccines, oncolytic viruses, sound wave therapy, magnetic therapy, electrical therapy, and electrostatic therapy. The method of any one of claims 1 to 3 and 5 to 21, wherein the proinflammatory agent comprises a TLR agonist. The method of claim 22, wherein the TLR agonist activates TLRs on macrophages, optionally wherein the TLRs comprise TLR2, TLR3, TLR7, TLR8 and/or TLR9. The method of claim 23, wherein the TLR agonist comprises CpG, poly I:C and/or R848. The method of any one of claims 1 to 3 and 5 to 24, wherein the pro-inflammatory agent comprises a bacterial component, optionally comprising lipopolysaccharide (LPS). The method of any one of claims 1 to 3 and 5 to 25, wherein the proinflammatory agent comprises a STING activator. The method of claim 26, wherein the STING activator comprises 2'3'-cGAMP. The method of any one of claims 1 to 3 and 5 to 27, wherein the pro-inflammatory agent comprises a chemotherapeutic agent. The method of claim 28, wherein the chemotherapy comprises azathioprine (AZA). The method of any one of claims 1 to 3 and 5 to 29, wherein the proinflammatory agent comprises a proinflammatory cytokine. The method of claim 30, wherein the pro-inflammatory interleukins comprise IL-1b, IL-18, IL-6 and/or TNFα. The method of any one of claims 1-2 and 5-31, wherein the pro-inflammatory agent comprises radiation therapy. The method of claim 32, wherein the radiation therapy comprises irradiating a site of the cancer to be treated. The method of claim 32 or claim 33, wherein the radiation therapy comprises irradiating a site different from the cancer site to be treated. The method of any one of claims 32 to 34, wherein the radiation therapy dose is insufficient to kill tumor cells. The method of any one of claims 1-2 and 5-35, wherein the proinflammatory agent comprises a checkpoint inhibitor. 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. The method of any one of claims 1 to 3 and 5 to 37, wherein the proinflammatory agent is administered intermittently. The method of any one of claims 1 to 3 and 5 to 38, wherein the proinflammatory agent and the SHP-1 inhibitor are administered simultaneously or concurrently. A method as claimed in any one of claims 1 to 3 and 5 to 39, wherein the pro-inflammatory agent comprises immune cells. The method of any one of claims 4 to 39, wherein the method further comprises an immune cell. The method of claim 41, wherein the immune cells are derived from the same individual. The method of claim 41 or claim 42, wherein the immune cells comprise or are macrophages, and optionally wherein the macrophages have an M1 phenotype. The method of any one of claims 40 to 43, wherein the immune cells are derived from monocytes. The method of any one of claims 40 to 44, wherein the immune cells express high levels of MHC-I, MHC-II, CD80 and/or CD86. The method of any one of claims 40 to 45, wherein the immune cells express one or more pro-inflammatory cytokines, optionally wherein the one or more pro-inflammatory cytokines comprise TNFα and/or IL-12. The method of any one of claims 40 to 46, wherein the immune cells do not express significant levels of TGFβ and/or IL-10. The method of any one of claims 40 to 47, wherein the immune cells comprise T cells. The method of any one of claims 40 to 48, wherein the immune cells are engineered to express a chimeric antigen receptor, optionally wherein the chimeric antigen receptor specifically binds to a tumor antigen. The method of any one of claims 43 to 49, wherein the macrophages are engineered to be defective in SHP-1 expression and/or activation. The method of any one of claims 40 to 50, wherein 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. The method of any one of claims 40 to 51, wherein the immune cells are administered simultaneously or concurrently with the SHP-1 inhibitor. The method of any one of claims 1 to 52, further comprising administering to the individual an effective amount of an anti-TNFα antibody. The method of any one of claims 1 to 53, wherein the cancer is a solid tumor. The method of any one of claims 1 to 53, wherein the cancer is a hematological cancer. The method of any one of claims 1 to 55, wherein the cancer is advanced cancer. The method of any one of claims 1 to 56, wherein the cancer is resistant or refractory to radiation therapy, chemotherapy, and/or checkpoint inhibitors. The method of any one of claims 1 to 57, wherein the individual is a human. A composition comprising a SHP-1 inhibitor and a proinflammatory agent, wherein the proinflammatory agent comprises an agent selected from the group consisting of immune cells, TLR agonists, STING activators, agents for radiation therapy, PAMP/DAMP activators, checkpoint inhibitors, proinflammatory cytokines, chemotherapeutic agents, bacterial components, cancer vaccines, oncolytic viruses, and agents for ultrasound therapy, magnetic therapy, electrical therapy or electrostatic therapy.