Compositions and methods for inhibiting expression of TMIGD2

Abstract

This disclosure provides methods of treating cancer by inhibiting expression and / or activity of TMIGD2. In some embodiments, the methods include administering (a) a TMIGD2 mRNA-targeting agent, (b) a gene-based therapeutic agent, (c) a TMIGD2 inhibitory small molecule, or (d) one or more TMIGD2 antibodies or antigen-binding fragments thereof to inhibit expression and / or activity of TMIGD2 in a subject in need thereof.

Description

[Technical field]

[0001] Priority This application claims priority to U.S. Provisional Patent Application No. 63 / 217,630 (filed July 1, 2021), the contents of which are incorporated by reference in their entirety.

[0002] Federal Funding Statement This invention was made with Government support under R01CA175495 awarded by the National Institutes of Health. The Government has certain rights in this invention.

[0003] Sequence Listing This application contains a Sequence Listing in accordance with ST.26 that has been submitted in xml format via EFS-Web and is incorporated herein by reference in its entirety. A copy of the xml format created on July 1, 2022 is named SequenceListing.xml and is 66.1KB in size. [Background technology]

[0004] Cancer is a serious public health problem in the United States and other countries. More than 90% of cancer deaths occur due to cancer metastasis and not the primary cancer. In the United States alone, there will be approximately 924,310 new cases of cancer and 339,150 deaths from cancer. According to Cancer Statistics 2010, in the United States alone, there will be an estimated 222,520 new cases of lung cancer and 157,300 deaths, 217,730 new cases of prostate cancer and 32,050 deaths, 207,090 new cases of breast cancer and 39,840 deaths, 145,500 new cases of intestinal cancer and 51,370 deaths, 58,240 new cases of kidney cancer and 8,210 deaths, and 51,350 new cases of pancreatic cancer and 36,800 deaths. Although conventional treatments such as surgery, chemotherapy and radiation therapy can often control the growth of the primary cancer, successful cancer control remains rare.

[0005] Thus, there is a serious and long-standing need for the development of more effective cancer treatments. Summary of the Invention

[0006] The invention provides a method of preventing or treating cancer in a subject in need thereof, the method comprising administering to the subject an effective amount of an agent that inhibits TMIGD2 expression, activity, or both.

[0007] In some aspects, the agent is selected from the group consisting of an antibody agent, an mRNA targeting agent, a small molecule agent, and a gene editing agent.

[0008] In some embodiments, the mRNA targeting agent is an antisense agent or an RNAi agent. In some embodiments, the antisense agent comprises or consists of a nucleic acid sequence complementary to the mRNA encoded by the nucleic acid sequence shown in SEQ ID NO:4, SEQ ID NO:5, or SEQ ID NO:6, or the antisense agent comprises or consists of a nucleic acid sequence having about 80%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5% or more sequence identity to the mRNA encoded by the nucleic acid sequence shown in SEQ ID NO:4, SEQ ID NO:5, or SEQ ID NO:6.

[0009] In some embodiments, the RNAi agent is selected from the group consisting of small interfering RNA (siRNA), double stranded RNA (dsRNA), microRNA (miRNA), piwiRNA (piRNA), small nucleolar RNA (snoRNA), small tRNA derived RNA (tsRNA), small regulatory RNA (srRNA) and small hairpin RNA (shRNA) molecules. In some embodiments, the RNAi agent comprises or consists of a nucleic acid sequence complementary to an mRNA encoded by a nucleic acid sequence set forth in SEQ ID NO:4, SEQ ID NO:5 or SEQ ID NO:6, or the RNAi agent comprises or consists of a nucleic acid sequence having about 80%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5% or more sequence identity to an mRNA encoded by a nucleic acid sequence set forth in SEQ ID NO:4, SEQ ID NO:5 or SEQ ID NO:6.

[0010] In some embodiments, the gene editing agent is selected from the group consisting of a TALEN-based agent, a ZFN-based agent, and a CRISPR-based agent. In some embodiments, the gene editing agent knocks out or knocks down the expression of TMIGD2.

[0011] In some embodiments, the antibody agent is an antibody or antigen-binding fragment thereof that specifically binds to an epitope within the extracellular domain of TMIGD2. In some embodiments, the extracellular domain of TMIGD2 comprises residues 1-150 of the amino acid sequence set forth in SEQ ID NO:1 or SEQ ID NO:2, or residues 1-30 of the amino acid sequence set forth in SEQ ID NO:3.

[0012] In some embodiments, the antibody or antigen-binding fragment thereof comprises: (a) a heavy chain variable region comprising GYTFTSYDIN (SEQ ID NO:24), WIYPGDGSTNYNEKFKG (SEQ ID NO:25), and / or ARRGLRYYFDY (SEQ ID NO:26); and (b) a light chain variable region comprising RASQDIRNYLN (SEQ ID NO:32), YTSRLHS (SEQ ID NO:33), and QQVNTLPWT (SEQ ID NO:34). In some embodiments, the antibody or antigen-binding fragment thereof comprises: (a) a heavy chain variable region comprising GYSITSDYAWN (SEQ ID NO:56), YITYSGSTSYNPSLKS (SEQ ID NO:57), and / or ARSGYRYDDAMDY (SEQ ID NO:58); and (b) a light chain variable region comprising KSSQSLLSSNNQKNYLA (SEQ ID NO:64), FASTRES (SEQ ID NO:65), and QQHYRTPLT (SEQ ID NO:66).

[0013] In some embodiments, the antibody or antigen-binding fragment thereof comprises: (a) a heavy chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 23 or an amino acid sequence having at least 85%, 90%, 95%, 99% or more sequence identity to the amino acid sequence set forth in SEQ ID NO: 23; and / or (b) a light chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 31 or an amino acid sequence having at least 85%, 90%, 95%, 99% or more sequence identity to the amino acid sequence set forth in SEQ ID NO: 31. In some embodiments, the antibody or antigen-binding fragment thereof comprises: (a) a heavy chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 55 or an amino acid sequence having at least 85%, 90%, 95%, 99% or more sequence identity to the amino acid sequence set forth in SEQ ID NO: 25; and / or (b) a light chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 63 or an amino acid sequence having at least 85%, 90%, 95%, 99% or more sequence identity to the amino acid sequence set forth in SEQ ID NO: 63.

[0014] In some embodiments, the cancer is a human hematological malignancy. In some embodiments, the human hematological malignancy is selected from the group consisting of myeloid neoplasms, acute myeloid leukemia (AML), AML with recurrent genetic abnormalities, AML with myelodysplasia-related changes, therapy-related AML, acute leukemia of unclear lineage, myeloproliferative neoplasms, essential thrombocythemia, polycythemia vera, myelofibrosis (MF), primary myelofibrosis, systemic mastocytosis, myelodysplastic syndromes (MDS), myeloproliferative / myelodysplastic syndromes, chronic my ... Myelogenous leukemia, chronic neutrophilic leukemia, chronic eosinophilic leukemia, myelodysplastic syndromes (MDS), refractory anemia with sideroblasts, refractory cytopenia with polycythemia dysplasia, refractory anemia with excess blasts (type 1), refractory anemia with excess blasts (type 2), MDS with isolated 5q deletion, unclassifiable MDS, myeloproliferative / myelodysplastic syndrome, chronic myelomonocytic leukemia, atypical chronic myelogenous leukemia, juvenile myelomonocytic leukemia, unclassifiable myeloproliferative Myelodysplastic / Myelodysplastic syndromes, Lymphoid neoplasms, Precursor lymphoid neoplasms, B-lymphoblastic leukemia, B-lymphoblastic lymphoma, T-lymphoblastic leukemia, T-lymphoblastic lymphoma, Mature B-cell neoplasms, Diffuse large B-cell lymphoma, Primary central nervous system lymphoma, Primary mediastinal B-cell lymphoma, Burkitt lymphoma / leukemia, Follicular lymphoma, Chronic lymphocytic leukemia, Small lymphocytic lymphoma, B-cell prolymphocytic leukemia, Lymphoplasmocytic leukemia and / or pulmonary leukemia, pulmonary cytoma, pulmonary cytoma (PCL), myeloma, cystic leukemia, myeloma, cystic leukemia, pulmonary fibrosis ...

[0015] In another aspect, the invention provides an anti-TMIGD2 antibody or antigen-binding fragment thereof. In some embodiments, the antibody or antigen-binding fragment thereof described herein comprises: (a) a heavy chain variable region comprising GYTFTSYDIN (SEQ ID NO: 24), WIYPGDGSTNYNEKFKG (SEQ ID NO: 25), and ARRGLRYYFDY (SEQ ID NO: 26); and a light chain variable region comprising RASQDIRNYLN (SEQ ID NO: 32), YTSRLHS (SEQ ID NO: 33), and QQVNTLPWT (SEQ ID NO: 34); or (b) a heavy chain variable region comprising GYSITSDYAWN (SEQ ID NO: 56), YITYSGSTSYNPSLKS (SEQ ID NO: 57), and ARSGYRYDDAMDY (SEQ ID NO: 58); and a light chain variable region comprising KSSQSLLSSNNQKNYLA (SEQ ID NO: 64), FASTRES (SEQ ID NO: 65), and QQHYRTPLT (SEQ ID NO: 66). In some embodiments, the antibodies or antigen-binding fragments thereof described herein comprise: (a) a heavy chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 23 or an amino acid sequence having at least 85%, 90%, 95%, 99% or more sequence identity to the amino acid sequence set forth in SEQ ID NO: 23; and (b) a light chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 31 or an amino acid sequence having at least 85%, 90%, 95%, 99% or more sequence identity to the amino acid sequence set forth in SEQ ID NO: 31. In some embodiments, the antibodies or antigen-binding fragments thereof described herein comprise: (a) a heavy chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 55 or an amino acid sequence having at least 85%, 90%, 95%, 99% or more sequence identity to the amino acid sequence set forth in SEQ ID NO: 25; and (b) a light chain variable region comprising an amino acid sequence set forth in SEQ ID NO: 63 or an amino acid sequence having at least 85%, 90%, 95%, 99% or more sequence identity to the amino acid sequence set forth in SEQ ID NO: 63. [Brief description of the drawings]

[0016] [Figure 1]Figures 1A-1B are representative images of flow cytometry showing that TMIGD2 is highly expressed in various human hematological malignancies including human erythroleukemia (HEL), chronic myeloid leukemia (CML) and acute myeloid leukemia (Kg1a) (Figure 1A). Histograms of anti-TMIGD2 mAb (open) and isotype control (shaded) are shown. From Cancer Cell Line Encyclopedia (CCLE) and Genevestigator, TMIGD2 mRNA was highly expressed in cell lines such as human leukemia, lymphoma, and multiple myeloma (Figure 1B). [Diagram 2] Figures 2A-2C are representative diagrams showing that TMIGD2, but not PD-L1 / PD-1, mRNA is highly expressed in human AML and associated with worse overall survival in patients (Figures 2A-2B). Interactive analysis of gene expression profiling was performed on the mRNA levels of the newly identified HHLA2 / TMIGD2 / KIR3DL3 pathway and the traditionally known PD-L1 / PD-1 pathway in the TCGA and GTEx datasets of human AML. 173 tumor data, 70 normal data; *P<0.05. (Figure 2C). In AML patients, the overall survival of the TMIGD2-high group (top 25%) was significantly (p=0.011) worse than that of the TMIGD2-low group (remaining 75%). (Figure 2C). [Diagram 3] Figures 3A-B are representative diagrams comparing the expression of TMIGD2 on stem / progenitor cells of AML patients with differentiating blast cells of AML patients (Figure 3A) or CD34+ normal stem / progenitor cells in cord blood / adult bone marrow mononuclear cells of healthy donors (Figure 3B). 40 AML patients, 10 healthy donors. **P<0.01, ****P<0.0001, determined by two-tailed paired (Figure 3A) or unpaired (Figure 3B) Student's t-test. Mean values ​​are shown unless otherwise stated, and error bars indicate ±SEM. [Figure 4]Figures 4A-4D are representative diagrams showing that TMIGD2 is enriched in functional leukemia-initiating cells. Figure 4A is a schematic diagram of flow cytometric sorting of TMIGD2+ and TMIGD2- AML stem cells followed by colony forming unit (CFU) assays and in vivo limiting dilution xenograft assays. Figure 4B shows the results of the first and second CFU assays with TMIGD2+ or TMIGD2- primary AML stem cells from patients #31 and #27. Figure 4C shows leukemic cell engraftment in irradiated NSG mice transplanted with TMIGD2+ and TMIGD2- AML stem cells from patient #31. Transcriptome-wide RNA sequencing (RNA-seq) was performed on flow-sorted CD34+TMIGD2+ and CD34+TMIGD2- fractions of six primary AML specimens. Figure 4D shows that gene set enrichment analysis (GSEA) showed that the top seven pathways enriched in the CD34+TMIGD2+ fraction consisted of E2F targets, MYC targets, and G2M checkpoints, consistent with the finding that CD34+TMIGD2+ cells generate more colonies and induce leukemia more efficiently than CD34+TMIGD2- cells. Furthermore, Figure 4D shows that the CD34+TMIGD2+ population is associated with established leukemia stem cells (LSCs) and a 17-gene stemness signature, and the CD34+TMIGD2- fraction correlates with myeloid cell development, hematopoietic maturation, and downregulation of HOXA9 and MEIS1 targets. [Diagram 5]Figures 5A-5D are representative figures showing that loss or blockage of TMIGD2 reduces the maintenance of AML stem cells. These figures include, among others, a schematic of the TMIGD2+ AML stem cell flow cytometry sorting strategy and lentiviral transduction (Figure 5A), FACS experiments of TMIGD2 expression in AML stem cells transduced with lentivirus expressing scrambled control shRNA (shCtrl) or TMIGD2-specific shRNA (shTMIGD2) (Figure 5B), and quantification of colony forming unit results in three AML patients (three independent experiments with shCtrl, shTMIGD2#2, and shTMIGD2#3) (Figure 5C). The therapeutic efficacy of anti-TMIGD2 mAbs 17C7 and 20F2 was evaluated in vivo using AML patient-derived xenografts (PDX) of various clinically relevant AML subtypes. The anti-leukemic effect of anti-TMIGD2 mAbs 17C7 and 20F2 was confirmed by the reduction of human CD45+ cells (AML cells) in peripheral blood and bone marrow after treatment (Figure 5D). *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, determined by two-tailed unpaired Student's t-test. Means are shown unless otherwise stated, and error bars indicate ±SEM. [Figure 6] Figures 6A and 6B are representative figures showing that knockdown of TMIGD2 increases cell death in human hematological malignancies. This figure specifically includes apoptosis analysis of shCtrl and shTMIGD2#3 HEL cells. Figure 6B shows early apoptosis, Annexin V+DAPI-, late apoptosis / necrosis, Annexin V+DAPI+, and Figure 6C shows representative heatmaps showing genes enriched in apoptosis and cell cycle arrest. *P<0.05, **P<0.01, ***P<0.001, as determined by two-tailed unpaired Student's t-test. Mean values ​​are shown unless otherwise stated, and error bars indicate ±SEM. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0017] Detailed Description The B7 ligand family binds to the CD28 receptor family on T cells and other immune cells and critically regulates immune cell function. 1,2 The B7 / CD28 pathway is an attractive therapeutic target, and the FDA has approved several drugs developed from the B7 / CD28 family. 3-6 HERV-H LTR-associated protein 2 (HHLA2) was discovered in 2013 as a new functional member of the B7 family. 7 This subsequently led to the discovery of a new member of the CD28 family, immunoglobulin domain-containing protein 2 (TMIGD2), as a receptor for HHLA2. 8,9 TMIGD2 is expressed on T cells and NK cells and exerts costimulatory functions on T cells and NK cells. 1,10,11 TMIGD2 has at least three isoforms: isoform 1 (SEQ ID NO: 1, NCBI NP_653216.2), isoform 2 (SEQ ID NO: 2, NCBI NP_001162597.1), and isoform 3 (SEQ ID NO: 3, NCBI NP_001295161.1). Representative DNA sequences encoding isoforms 1 to 3 are shown in SEQ ID NO: 4 (NCBI NM_144615), SEQ ID NO: 5 (NCBI NM_001169126.1), and SEQ ID NO: 6 (NCBI NM_001308232), respectively.

[0018] [Table 1]

[0019] [Table 2]

[0020] As disclosed herein, it has been found that TMIGD2 is expressed in various human hematological malignancies, is functionally important for leukemia-initiating cells, and is associated with poor overall survival in AML patients. Knocking down TMIGD2 expression has been found to reduce AML stem cell maintenance and increase cell death in human hematological malignancies. In some embodiments, treatment with anti-TMIGD2 monoclonal antibodies inhibits AML progression in vivo. Based on these findings, the present disclosure provides methods of treating hematological malignancies using one or more agents that inhibit TMIGD2 expression and / or activity, agents and kits for use in the methods, and the use of the agents and kits to inhibit TMIGD2 expression and / or activity. Exemplary agents that inhibit TMIGD2 expression and / or activity include, but are not limited to, mRNA targeting agents such as antisense agents or RNAi agents, gene editing agents such as TALEN, ZFN or CRISPR-based gene editing agents, small molecules, antagonist antibodies and fusion proteins thereof, and TMIGD2-binding peptides.

[0021] While this disclosure can be embodied in various forms, the following description of several embodiments is provided with the understanding that this disclosure should be considered as an example of the invention and is not intended to limit the invention to the specific embodiments shown. Headings are provided for convenience only and should not be construed as limiting the invention in any manner. Embodiments shown under any heading can be combined with embodiments shown under other headings.

[0022] definition Numerical values ​​for various quantitative values ​​specified in this application are described as approximations, unless otherwise indicated, as if the word "about" preceded both the minimum and maximum values ​​in the stated range. It is to be understood, although not always explicitly stated, that all numerical values ​​are preceded by the term "about." It is to be understood that such range formats are used for convenience and brevity, and should be understood to be flexible, including not only the numerical values ​​explicitly specified as the limits of the range, but also to include all individual numerical values ​​or subranges subsumed within the range, as if each numerical value and subrange were explicitly specified. For example, a range of about 1 to about 200 should be understood to include the explicitly recited limits of about 1 and about 200, but also individual ones such as about 2, about 3, and about 4, as well as subranges such as about 10 to about 50, about 20 to about 100, etc. It is also to be understood, although not always explicitly stated, that the reagents described in the specification are merely exemplary, and equivalents thereof are known in the art.

[0023] The term "about" as used herein when referring to a measurable value, such as an amount or concentration, is meant to include variations of 20%, 10%, 5%, 1%, 0.5%, and even 0.1% of the stated amount.

[0024] As used herein, an agent that "inhibits" TMIGD2 expression and / or activity reduces TMIGD2 expression and / or activity by at least 5% relative to TMIGD2 expression and / or activity in the absence of the agent. In certain embodiments, an agent may reduce TMIGD2 expression and / or activity by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 99% or 100% (i.e., complete inhibition) relative to TMIGD2 expression and / or activity in the absence of the agent.

[0025] The term "antibody" as used herein refers to an immunoglobulin molecule or immunologically active portion thereof that binds to a specific antigen (e.g., TMIGD2). In embodiments where the antibody used in the methods, compositions, and kits is a full-length immunoglobulin molecule, the antibody comprises two heavy chains and two light chains, each of which has three complementarity determining regions (CDRs). In embodiments where the antibody is an immunologically active portion of an immunoglobulin molecule, the antibody may be, for example, a Fab, Fab', Fv, Fab', F(ab')2, a disulfide-linked Fv, scFv, a single domain antibody (dAb), or a diabody. The antibodies used in the methods, compositions, and kits may include natural antibodies, synthetic antibodies, monoclonal antibodies, polyclonal antibodies, chimeric antibodies, humanized antibodies, multispecific antibodies, bispecific antibodies, dual-specific antibodies, anti-idiotypic antibodies, or fragments thereof that retain the ability to bind to a specific antigen (e.g., TMIGD2).

[0026] The term "RNAi" in this specification refers to interfering RNA or RNA interference. RNAi refers to a means of selective post-transcriptional gene silencing by the destruction of specific mRNAs by molecules that bind to and inhibit their processing, for example, by inhibiting mRNA translation or by degrading the mRNA molecule. The term "RNAi" in this specification refers to interfering RNAs, including but not limited to siRNAi, shRNAi, endogenous microRNAs, and artificial microRNAs. For example, it includes sequences previously identified as siRNAs, regardless of the mechanism of downstream processing of the RNA. (i.e., siRNAs are believed to have a specific in vivo processing method that causes cleavage of mRNAs, but such sequences can be incorporated into vectors in conjunction with flanking sequences described in the specification.)

[0027] The ranges described herein are intended as continuous ranges, including all values ​​between the minimum and maximum values ​​recited, and any ranges formed by such values. All ratios (and ranges of such ratios) formed by dividing a disclosed numerical value by another disclosed numerical value are also disclosed herein. Thus, one of ordinary skill in the art will appreciate that many such ratios, ranges, and ratio ranges can be clearly derived from the numerical values ​​set forth herein, and in all cases, such ratios, ranges, and ratio ranges represent various aspects of this disclosure.

[0028] method The present disclosure provides a method of treating a condition responsive to TMIGD2 inhibition in a subject in need thereof, comprising administering to the subject an agent that inhibits expression and / or activity of TMIGD2. In some embodiments, the agent can be an antibody agent, an mRNA targeting agent (e.g., an antisense agent or an RNAi agent), a small molecule agent, a gene editing agent (e.g., a TALEN-based agent, a ZFN-based agent, a CRISPR-based agent), or a polypeptide agent. In some embodiments, administration of the agent enhances an immune response.

[0029] In some embodiments, the condition responsive to TMIGD2 inhibition is cancer. In some of these embodiments, the cancer is selected from the group consisting of chronic lymphocytic leukemia (CLL), acute leukemia, acute lymphocytic leukemia (ALL), B-cell acute lymphocytic leukemia (B-ALL), T-cell acute lymphocytic leukemia (T-ALL), T-cell lymphoma, B-cell lymphoma, chronic myelogenous leukemia (CML), acute myelogenous leukemia, B-cell prolymphocytic leukemia, blastic plasmacytoid dendritic cell neoplasm, Burkitt's lymphoma, diffuse large B-cell lymphoma, and leukemia. cell lymphoma, follicular lymphoma, hairy cell leukemia, small cell follicular lymphoma, large cell follicular lymphoma, malignant lymphoproliferative disorders, MALT lymphoma, mantle cell lymphoma, marginal zone lymphoma, multiple myeloma, myelodysplasia and myelodysplastic syndromes, non-Hodgkin's lymphoma, Hodgkin's lymphoma, plasmablastic lymphoma, plasmacytoid dendritic cell neoplasm, Waldenstrom's macroglobulinemia, or preleukemia.In other aspects, the cancer is a myeloid neoplasm, acute myeloid leukemia (AML), AML with recurrent genetic abnormalities, AML with myelodysplasia-related changes, therapy-related AML, acute leukemia of unclear lineage, myeloproliferative neoplasms, essential thrombocythemia, polycythemia vera, myelofibrosis (MF), primary myelofibrosis, systemic mastocytosis, myelodysplastic syndrome (MDS), myeloproliferative / myelodysplastic syndrome, chronic myeloid leukemia, chronic Neutrophilic leukemia, chronic eosinophilic leukemia, myelodysplastic syndromes (MDS), refractory anemia with sideroblasts, refractory cytopenia with polycythemia dysplasia, refractory anemia with excess blasts (type 1), refractory anemia with excess blasts (type 2), MDS with isolated 5q deletion, unclassifiable MDS, myeloproliferative / myelodysplastic syndrome, chronic myelomonocytic leukemia, atypical chronic myelogenous leukemia, juvenile myelomonocytic leukemia, unclassifiable myeloproliferative / myelodysplastic syndrome , lymphoid neoplasms, precursor lymphoid neoplasms, B-lymphoblastic leukemia, B-lymphoblastic lymphoma, T-lymphoblastic leukemia, T-lymphoblastic lymphoma, mature B-cell neoplasms, diffuse large B-cell lymphoma, primary central nervous system lymphoma, primary mediastinal B-cell lymphoma, Burkitt's lymphoma / leukemia, follicular lymphoma, chronic lymphocytic leukemia, small lymphocytic lymphoma, B-cell prolymphocytic leukemia, lymphoplasmacytic lymphoma, Waldemarcated lymphoma, Human hematological malignancies such as Ström's macroglobulinemia, mantle cell lymphoma, marginal zone lymphoma, post-transplant lymphoproliferative disease, HIV-associated lymphoma, primary effusion lymphoma, intravascular large B-cell lymphoma, primary cutaneous B-cell lymphoma, hairy cell leukemia, multiple myeloma, monoclonal gammopathy of undetermined significance (MGUS), smoldering multiple myeloma or solitary plasmacytoma (bone and extramedullary).

[0030] In some embodiments, the cancer is selected from the group consisting of adrenal gland cancer, anal cancer, basal and squamous cell skin cancer, bile duct cancer, bladder cancer, bone cancer, brain and spinal cord tumors, breast cancer, cervical cancer, colorectal cancer, endometrial cancer, esophageal cancer, Ewing's family of tumors, eye cancer (ocular melanoma), gallbladder cancer, gastrointestinal neuroendocrine (carcinoid) tumors, gastrointestinal stromal tumors (GIST), gestational trophoblastic disease, Kaposi's sarcoma, kidney cancer, laryngeal and hypopharyngeal cancer, liver cancer, lung cancer, pulmonary carcinoid tumors, malignant mesothelioma, melanoma skin cancer, Merkel cell skin cancer, cancer of the nasal cavity and paranasal sinuses, Nasopharyngeal carcinoma, neuroblastoma, non-small cell lung cancer, tumors of the central nervous system (CNS), cancer of the oral cavity and oropharynx, osteosarcoma, ovarian cancer, pancreatic cancer, pancreatic neuroendocrine tumors (NETs), penile cancer, pituitary tumors, prostate cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, skin cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, gastric cancer, testicular cancer, thymic cancer, thyroid cancer, uterine sarcoma, vaginal cancer, vulvar cancer, Waldenstrom's macroglobulinemia, Wilms' tumor, squamous cell carcinoma, environmentally induced cancer, combinations of cancers, and metastatic lesions of cancer. In some embodiments, the cancer is a leukemia or lymphoma, such as lymphoblastic lymphoma or B-cell non-Hodgkin's lymphoma.

[0031] In certain embodiments, the methods provided herein further include administration of a second agent. In certain of these embodiments, the second agent also inhibits expression and / or activity of TMIGD2. In other embodiments, the second agent is an agent other than a TMIGD2 inhibitor used in the treatment of a condition, including, for example, radiation therapy or chemotherapy. In these combination embodiments, the first and second agents may be administered simultaneously or sequentially, by the same or different routes. When the two agents are administered simultaneously, they may be administered in a single formulation or in separate formulations. When the two agents are administered sequentially, the interval between administration of the two agents may be the same or different. For example, one agent may be administered more frequently than the other agent, and one agent may be administered for a longer period of time. In certain embodiments, the second agent may be administered one or more hours, one or more days, or one or more weeks after administration of the first agent, or vice versa. In certain embodiments, the first agent may be administered one or more times prior to the first administration of the second agent. When a second agent is administered, administration of the first agent may be discontinued or continued for all or part of the period during which the second agent is administered.

[0032] A. Antibody drugs In certain aspects of the methods provided herein, the agent that inhibits the expression and / or activity of TMIGD2 is an antibody or an antigen-binding fragment thereof, or a fusion protein thereof.

[0033] In these embodiments, the antibody or antigen-binding fragment thereof specifically binds to an epitope of TMIGD2 (e.g., TMIGD2 isoform 1, 2, or 3 of the sequences set forth in SEQ ID NOs: 1-3, respectively). In some embodiments, the antibody or antigen-binding fragment thereof cross-reacts with more than one TMIGD2 isoform, while in other embodiments, the antibody is specific for one isoform. For example, in some embodiments, the antibody or antigen-binding fragment thereof may bind to both isoforms 1 and 2 and not isoform 3, or vice versa. In some embodiments, the antibody or antigen-binding fragment thereof binds only to human TMIGD2. In other embodiments, the antibody or antigen-binding fragment thereof binds to non-human TMIGD2 (e.g., murine TMIGD2) in addition to or instead of human TMIGD2. In some embodiments, the antibody or antigen-binding fragment thereof partially or completely blocks binding of TMIGD2 to HHLA. In some embodiments, the antibody or antigen-binding fragment thereof modulates (e.g., inhibits) one or more aspects of TMIGD2 signaling (e.g., TMIGD2 phosphorylation).

[0034] In certain embodiments, the antibody or antigen-binding fragment thereof binds to an epitope that is present, in whole or in part, in the extracellular domain of TMIGD2 (e.g., residues 1-150 of the amino acid sequence set forth in SEQ ID NO: 1 or 2, or residues 1-30 of the amino acid sequence set forth in SEQ ID NO: 3). In some of these embodiments, the antibody or antigen-binding fragment thereof binds to the extracellular domain of all TMIGD2 isoforms. In other embodiments, the antibody or antigen-binding fragment thereof is specific for one or more isoforms. For example, the antibody or antigen-binding fragment thereof may bind to the extracellular domain of isoforms 1 and 2 but not the extracellular domain of isoform 3, or vice versa.

[0035] In some embodiments, the antibody or antigen-binding fragment thereof is a one-arm antibody (an antibody in which the heavy chain variable domain and the light chain variable domain form a single antigen-binding arm) having an Fc region, wherein the Fc region comprises a first and a second Fc polypeptide, wherein the first and second Fc polypeptides are in a complex to form an Fc region that increases the stability of the antibody fragment compared to a Fab molecule having the antigen-binding arm.

[0036] In one embodiment, the antibody or antigen-binding fragment thereof is a chimeric antibody, e.g., an antibody having antigen-binding sequences (e.g., framework sequences and / or constant domain sequences) from a non-human donor grafted with xenogeneic non-human, human or humanized sequences. In one embodiment, the non-human donor is a mouse. In a further embodiment, the antigen-binding sequences are synthetic, e.g., obtained by mutagenesis (e.g., phage display screening, etc.). In a particular embodiment, the chimeric antibody of the invention has a mouse V region and a human C region. In one embodiment, the mouse light chain V region is fused to a human kappa light chain. In another embodiment, the mouse heavy chain V region is fused to a human IgG1 C region.

[0037] In some embodiments, the antibodies or antigen-binding fragments thereof described herein are or include (i) chimeric, human or humanized antibodies, or antigen-binding fragments thereof; (ii) monospecific or bispecific antibodies, or antigen-binding fragments thereof; and / or (iii) monoclonal antibodies or antigen-binding fragments thereof. In some embodiments, the antibodies or antigen-binding fragments thereof described herein may be or include other immunologically binding moieties known in the art, including, but not limited to, immunoglobulins, heavy chain antibodies, light chain antibodies or other protein scaffolds with antibody-like properties, and Fab fragments, Fab' fragments, F(ab')2 fragments, Fv fragments, disulfide-linked Fv fragments, scFv fragments, diabodies, triabodies, tetrabodies, minibodies, maxibodies, tandabs, BiTe, nanobodies, camelid antibodies, or combinations thereof. In some aspects, the antibody or antigen-binding fragment thereof is, or comprises, (i) a heavy chain constant region chosen from IgG1, IgG2, IgG3, or IgG4, and / or (ii) a light chain constant region chosen from a κ light chain or a λ light chain constant region.

[0038] In some embodiments, the antibody or antigen-binding fragment thereof is, or comprises, (a) a heavy chain variable region (VH) having one, two or three VH CDR sequences, each of which has at least about 90% sequence identity to a VH CDR listed in Table 3 or Table 4; and / or (b) a light chain variable region (VL) having one, two or three VL CDR sequences, each of which has at least about 90% sequence identity to a VL CDR listed in Table 3 or Table 4. In some embodiments, the antibody or antigen-binding fragment thereof is, or comprises, (a) a VH having one, two or three VH CDR sequences, each having at least about 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more sequence identity to the VH CDRs of Table 3 or Table 4; and / or (b) a VL having one, two or three VL CDR sequences, each having at least about 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more sequence identity to the VL CDRs of Table 3 or Table 4. In some embodiments, the antibody or antigen-binding fragment thereof is, or comprises: (a) a VH comprising one, two or three VH CDR sequences comprising or consisting of the VH CDRs of Table 3 or Table 4, respectively; and / or (b) a VL comprising one, two or three VL CDR sequences comprising or consisting of the VL CDRs of Table 3 or Table 4, respectively.

[0039] In some embodiments, the antibody or antigen-binding fragment thereof is, or comprises: (a) a VH having at least about 90% or more sequence identity to a VH of Table 3 or Table 4; and / or (b) a VL having at least about 90% or more sequence identity to a VL of Table 3 or Table 4. In some embodiments, the antibody or antigen-binding fragment thereof is, or comprises: (a) a VH having at least about 95%, 96%, 97%, 98%, 99%, 99.5% or more sequence identity to a VH of Table 3 or Table 4; and / or (b) a VL having at least about 95%, 96%, 97%, 98%, 99%, 99.5% or more sequence identity to a VL of Table 3 or Table 4. In some embodiments, the antibody or antigen-binding fragment thereof is, or comprises, (a) a VH that comprises, or consists of, a VH of Table 3 or Table 4; and / or (b) a VL that comprises, or consists of, a VL of Table 3 or Table 4.

[0040] In some embodiments, the antibody or antigen-binding fragment thereof is, or comprises: (a) a heavy chain having at least about 90% or more sequence identity to a heavy chain of Table 3 or Table 4; and / or (b) a light chain having at least about 90% or more sequence identity to a light chain of Table 3 or Table 4. In some embodiments, the antibody or antigen-binding fragment thereof is, or comprises: (a) a heavy chain having at least about 95%, 96%, 97%, 98%, 99%, 99.5% or more sequence identity to a heavy chain of Table 3 or Table 4; and / or (b) a light chain having at least about 95%, 96%, 97%, 98%, 99%, 99.5% or more sequence identity to a light chain of Table 3 or Table 4. In some embodiments, the antibody or antigen-binding fragment thereof is, or comprises, (a) a heavy chain comprising, or consisting of, a heavy chain of Table 3 or Table 4; and / or (b) a light chain comprising, or consisting of, a light chain of Table 3 or Table 4.

[0041] In some embodiments, the antibodies or antigen-binding fragments thereof described herein are conjugated to a cytotoxic agent. In such embodiments, the cytotoxic agent is selected from the group consisting of therapeutic agents (e.g., chemotherapeutic agents), biological agents, toxins, and radioisotopes. Exemplary cytotoxic agents include taxol, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, doxorubicin, daunorubicin, dihydroxyanthracenedione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, and puromycin, as well as analogs or homologs thereof. Therapeutic agents include, but are not limited to, antimetabolites (e.g., methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine or 5-fluorouracil, decarbazine), alkylating agents (e.g., mechlorethamine, thiotepa, chlorambucil, melphalan, carmustine (BSNU) and lomustine (CCNU), cyclophosphamide, busulfan, dibromomannitol, streptozotocin, mitomycin C, and cis-dichlorodiamineplatinum(II) (DDP), cisplatin), anthracyclines (e.g., daunorubicin (formerly daunomycin) and doxorubicin), antibiotics (e.g., dactinomycin (formerly actinomycin), bleomycin, mithramycin, and anthramycin (AMC)), and antimitotic agents (e.g., vincristine and vinblastine). The antibodies or antigen-binding fragments thereof described herein can be conjugated with radioisotopes (e.g., radioactive iodine) to produce cytotoxic radiopharmaceuticals for treating related diseases such as cancer as described herein.

[0042] Antibody conjugates can be used to modify biological responses. The therapeutic moiety should not be construed as limited to classical chemical therapeutic agents. For example, the drug moiety may be a protein or polypeptide having a desired biological activity. Such proteins may include, for example, enzymatically active toxins such as abrin, ricin A, pseudomonas exotoxin, or diphtheria toxin, or active fragments thereof, proteins such as tumor necrosis factor or interferon gamma, or biological response modifiers such as lymphokines, interleukin-1 (IL-1), interleukin-2 (IL-2), interleukin-6 (IL-6), granulocyte macrophage colony stimulating factor (GM-CSF), granulocyte colony stimulating factor (G-SCF), or other cytokines or growth factors.Techniques for conjugating such therapeutic moieties to antibodies are well known, see, e.g., Arnon et al, “Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy”, in Monoclonal Antibodies And Cancer Therapy, Reisfeld et al. (eds.), pp. 243 56 (Alan R. Liss, Inc. 1985);Hellstrom et al, “Antibodies For Drug Delivery”, in Controlled Drug Delivery (2nd Ed.), Robinson et al. (eds.), pp. 623 53 (Marcel Dekker, Inc. 1987);Thorpe, “Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review”, in Monoclonal Antibodies '84: Biological And Clinical Applications, Pinchera et al. (eds.), pp. 475 506 (1985);“Analysis, Results, And Future Prospective Of The Therapeutic Else Of Radiolabeled Antibody In Cancer Therapy”, in Monoclonal Antibodies For Cancer Detection And Therapy, Baldwin et al. (eds.), pp. 303 16 (Academic Press 1985) and Thorpe et al., “The Preparation And Cytotoxic Properties Of Antibody-Toxin Conjugates”, Immunol. Rev., 62: 119 58 (1982).

[0043] In some embodiments, conjugation can be performed using a "cleavable linker" that facilitates the release of the cytotoxic agent or growth inhibitory agent inside the cell. For example, an acid-labile linker, a peptidase-sensitive linker, a photolabile linker, a dimethyl linker, or a disulfide-containing linker (see, e.g., U.S. Pat. No. 5,208,020) can be used. Alternatively, a fusion protein having an antibody or antigen-binding fragment thereof and a cytotoxic agent or a growth inhibitory agent can be produced by recombinant techniques or peptide synthesis. The DNA can contain respective regions encoding the two parts of the conjugate, adjacent to each other or separated by a region encoding a linker peptide that does not destroy the desired properties of the conjugate.

[0044] In some embodiments, the antibody or antigen-binding fragment thereof is the 17C7 monoclonal antibody or comprises the heavy and / or light chain sequences, the heavy and / or light chain variable sequences, or one or more CDR sequences of the 17C7 monoclonal antibody. The sequence of the 17C7 antibody is shown in Table 3.

[0045] [Table 3-1]

[0046] [Table 3-2]

[0047] In some embodiments, the antibody or antigen-binding fragment thereof is the 20F2 monoclonal antibody or comprises the heavy and / or light chain sequences, the heavy and / or light chain variable sequences, or one or more CDR sequences of the 20F2 monoclonal antibody. The sequence of the 20F2 antibody is shown in Table 4.

[0048] [Table 4-1]

[0049] [Table 4-2]

[0050] B. mRNA targeting drugs In certain aspects of the methods provided herein, the agent that inhibits expression and / or activity of TMIGD2 is an mRNA-targeting agent, such as an antisense agent or an RNAi agent.

[0051] In some aspects of the methods provided herein, the mRNA targeting agent is an antisense agent. Antisense agents (usually small fragments of DNA or RNA) bind to target mRNAs that code for proteins and form hybrid duplexes, thereby regulating protein expression. In cells, the antisense agent / mRNA hybrid is cleaved by ribonuclease H (RNAse H). Cleavage of the RNA strand from the duplex by RNAse H prevents the mRNA from being translated into protein.

[0052] In some aspects of the methods provided herein, the mRNA targeting agent is an RNAi agent. RNA interference (RNAi) is an evolutionarily conserved process in which expression or introduction of an RNA with a sequence identical or closely similar to that of a target gene results in sequence-specific degradation or specific post-transcriptional gene silencing (PTGS) of the messenger RNA (mRNA) transcribed from the target gene, thereby inhibiting expression of the target gene. RNAi agents typically consist of a nucleic acid or nucleic acid analog sequence specific to a target gene (e.g., TMIGD2). In some aspects, the RNAi agent is or includes a small nucleic acid, such as a small interfering RNA (siRNA), double-stranded RNA (dsRNA), microRNA (miRNA), piwiRNA (piRNA), small nucleolar RNA (snoRNA), small tRNA-derived RNA (tsRNA), small regulatory RNA (srRNA), or small hairpin RNA (shRNA) molecule. An siRNA agent refers to a nucleic acid that forms a double-stranded RNA that has the ability to reduce or inhibit the expression of the TMIGD2 gene when the siRNA is present or expressed in a cell in which the TMIGD2 gene is present. shRNA is a type of siRNA that functions similarly to RNAi and / or siRNA, but differs in that the shRNA has a double-stranded hairpin structure for increased stability. In some embodiments, the shRNA agent reduces or inhibits the expression of the TMIGD2 gene when the shRNA is present or expressed in a cell in which the TMIGD2 gene is present. In yet another such embodiment, the shRNA increases the expression of genes involved in apoptosis and cell cycle arrest. miRNAs are endogenous RNAs, some of which are known to regulate the expression of protein-coding genes at the post-transcriptional level. Endogenous microRNAs are small RNAs that naturally occur in the genome that can regulate the productive use of mRNA. In some embodiments, the miRNA agent is an artificial miRNA agent that contains an RNA sequence other than an endogenous microRNA that can regulate the productive use of mRNA. A dsRNA agent is an RNA molecule that is composed of two strands.dsRNA agents include RNA molecules that consist of a single RNA molecule that folds back on itself to form a double-stranded structure. For example, the stem-loop structure of the precursor molecule from which a single-stranded miRNA is derived is called a pre-miRNA and constitutes a dsRNA molecule. A piRNA agent refers to a nucleic acid molecule that has the ability to form an RNA-protein complex by interaction with an Argonaute protein of the piwi subfamily and reduce or inhibit the expression of the TMIGD2 gene when the piRNA is present or expressed in a cell in which the TMIGD2 gene is present. A snoRNA agent refers to a nucleic acid molecule that has the ability to guide the chemical modification of other RNAs and reduce or inhibit the expression of the TMIGD2 gene when the snoRNA is present or expressed in a cell in which the TMIGD2 gene is present.

[0053] In some embodiments, the mRNA targeting agent used in the methods provided herein comprises or consists of a nucleic acid sequence complementary to all or a portion of the TMIGD2 mRNA sequence. For example, in some embodiments, the mRNA targeting agent comprises or consists of a nucleic acid sequence complementary to all or a portion of the TMIGD2 mRNA encoded by the sequence shown in SEQ ID NO: 4 (a representative DNA sequence of TMIGD2 isoform 1), the sequence shown in SEQ ID NO: 5 (a representative DNA sequence of TMIGD2 isoform 2), or the sequence shown in SEQ ID NO: 6 (a representative DNA sequence of TMIGD2 isoform 3). In certain of these embodiments, the mRNA targeting agent may be complementary to a specific region of the TMIGD2 mRNA. For example, in one embodiment, the mRNA targeting agent is a TMIGD2 mRNA targeting agent that is a portion of the TMIGD2 mRNA corresponding to the extracellular domain of TMIGD2 (e.g., an mRNA corresponding to residues 1 to 150 of the amino acid sequence shown in SEQ ID NO: 1 or 2, or residues 1 to 30 of the amino acid sequence shown in SEQ ID NO: 3, including an mRNA encoded by nucleotides 1 to 450 of the nucleotide sequence shown in SEQ ID NO: 4 or 5, or nucleotides 1 to 90 of the nucleotide sequence shown in SEQ ID NO: 6), a portion of the TMIGD2 mRNA that is a transmembrane domain of TMIGD2 (e.g., an mRNA corresponding to residues 151 to 171 of the amino acid sequence shown in SEQ ID NO: 1 or 2, or residues 31 to 51 of the amino acid sequence shown in SEQ ID NO: 3, including an mRNA encoded by nucleotides 451 to 513 of the nucleotide sequence shown in SEQ ID NO: 4 or 5, or nucleotides 91 to 153 of the nucleotide sequence shown in SEQ ID NO: 6), or a TMIGD2 mRNA targeting agent that is a portion of the TMIGD2 mRNA that is a transmembrane domain of TMIGD2 (e.g., an mRNA corresponding to residues 151 to 171 of the amino acid sequence shown in SEQ ID NO: 1 or 2, or residues 31 to 51 of the amino acid sequence shown in SEQ ID NO: 3, including an mRNA encoded by nucleotides 451 to 513 of the nucleotide sequence shown in SEQ ID NO: 4 or 5, or nucleotides 91 to 153 of the nucleotide sequence shown in SEQ ID NO: 6). It is complementary to a portion of an mRNA (e.g., an mRNA corresponding to residues 172 to 282 of the amino acid sequence shown in SEQ ID NO: 1, residues 172 to 278 of the amino acid sequence shown in SEQ ID NO: 2, or residues 52 to 162 of the amino acid sequence shown in SEQ ID NO: 3, including an mRNA encoded by nucleotides 514 to 849 of the base sequence shown in SEQ ID NO: 4, nucleotides 514 to 837 of the base sequence shown in SEQ ID NO: 5, or nucleotides 514 to 489 of the base sequence shown in SEQ ID NO: 6).

[0054] It is understood in the art that a nucleic acid molecule does not need to be 100% complementary to a target nucleic acid sequence in order to specifically hybridize to the target sequence.Thus, in some embodiments, the mRNA targeting agent used in the methods provided herein may be at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 99% or 100% complementary to all or a portion of the TMIGD2 mRNA target sequence.

[0055] In some embodiments, the mRNA targeting agent used in the methods provided herein may be essentially completely complementary to all or part of the TMIGD2 mRNA sequence.The mRNA targeting agent of the present invention and the TMIGD2 mRNA are essentially completely complementary to each other when the degree of complementarity allows stable and specific binding between the mRNA targeting agent and the TMIGD2 mRNA.In some embodiments, an mRNA targeting agent that has one or two non-complementary nucleobases to the TMIGD2 mRNA is considered to be essentially completely complementary.

[0056] In one embodiment, an mRNA targeting agent of the invention and a target nucleic acid of TMIGD2 are fully complementary to each other. An mRNA targeting agent and a target nucleic acid of TMIGD2 are fully complementary to each other if each nucleobase of the mRNA targeting agent is complementary to the same number of nucleobases at corresponding positions of the target nucleic acid.

[0057] In some embodiments, the mRNA targeting agents used in this method have a backbone of linked monomeric subunits, each linked monomeric subunit being directly or indirectly bound to a heterocyclic base. The linkages linking the monomeric subunits, the sugar or sugar surrogate, and the heterocyclic base can be independently modified to give rise to multiple motifs of antisense agents, including hemimers, gapmers, alternations, uniform modifications, and positional modifications.

[0058] In one aspect, an mRNA targeting agent of the invention is 10-30 nucleosides in length, e.g., 15-30 linked or consecutive nucleosides, 10-25 linked or consecutive nucleosides, 20-30 linked or consecutive nucleosides, or 15-20 linked or consecutive nucleosides.

[0059] Delivery of the mRNA targeting agent can be achieved by any suitable method known in the art, including, but not limited to, viral vector delivery (e.g., lentiviral vector delivery, AAV viral vector delivery, adenoviral vector delivery), dendrimer-mediated delivery (e.g., dendrimer-based nanoparticle delivery), nanoparticle-mediated delivery, or combinations thereof.

[0060] C. Small molecule drugs In certain aspects of the methods provided herein, the agent that inhibits the expression and / or activity of TMIGD2 is a small molecule agent.

[0061] Small molecule anticancer drugs have been successfully used to target extracellular cell surface ligand-binding receptors and intracellular proteins, including anti-apoptotic proteins that play important roles in downstream signaling promoting cell proliferation and metastasis.

[0062] In some embodiments, the small molecule agent interferes with the activity of TMIGD2 by binding to and inhibiting activity of the TMIGD2 protein (e.g., TMIGD2 isoforms 1, 2, and / or 3). In certain of these embodiments, the small molecule binds to the extracellular domain of TMIGD2 (e.g., residues 1-150 of the amino acid sequence set forth in SEQ ID NO:1 or 2, or residues 1-30 of the amino acid sequence set forth in SEQ ID NO:3). In some embodiments, the small molecule agent reduces dimerization and / or aggregation of TMIGD2. In some embodiments, the small molecule agent partially or completely blocks binding of TMIGD2 to HHLA.

[0063] In some embodiments, the small molecule agent interferes with TMIGD2 expression and / or activity by binding to and inhibiting a protein involved in TMIGD2 expression (e.g., an upstream effector of TMIGD2, or a protein or transcription factor involved in TMIGD2 expression). In other embodiments, the small molecule agent interferes with TMIGD2 expression and / or activity by binding to a TMIGD2 nucleic acid (e.g., a TMIGD2 DNA or mRNA sequence).

[0064] D. Gene Editing Drugs In certain aspects of the methods provided herein, the agent that inhibits expression and / or activity of TMIGD2 is a gene editing agent.

[0065] Gene editing agents include agents that include one or more DNA or RNA sequences. In some embodiments, a gene editing agent includes multiple components. For example, a gene editing agent may include multiple vectors encoding various components (e.g., one or more gRNA sequences and one or more nucleases or nucleic acid sequences encoding nucleases).

[0066] In some embodiments, the gene editing agent modifies the sequence of the TMIGD2 gene or the sequence of a regulatory element (e.g., promoter or enhancer element) associated with the TMIGD2 gene to inhibit expression and / or activity of TMIGD2. For example, the gene editing agent can introduce a deletion, insertion, or mutation into the TMIGD2 gene or its regulatory element to knock out or knock down expression of TMIGD2. A gene is considered to be knocked out if the gene is completely removed or completely inactivated or suppressed by genetic manipulation. A gene is considered to be knocked down if the gene is partially inactivated or suppressed.

[0067] In some embodiments, the gene editing agent can introduce a genetic abnormality that completely blocks the expression of TMIGD2, for example, by disrupting the start codon of the TMIGD2 gene or a key regulatory element thereof. In other embodiments, the genetic abnormality can reduce the expression of TMIGD2 or express a truncated, inactive, or partially inactive form of TMIGD2, for example, by introducing a nonsense mutation into the gene, disrupting one or more exon sequences of the gene, and / or modifying one or more nucleotides that code for a functional domain or element of TMIGD2. In some embodiments, the agent introduces an inactivating mutation into the TMIGD2 gene. In some embodiments, the agent suppresses the transcription of the TMIGD2 gene. In some embodiments, the genetic abnormality reduces the expression or activity of TMIGD2 by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 99%, or 100% compared to the expression or activity in the absence of the abnormality. In some embodiments, the gene editing agent deletes or modifies one or more nucleotides located in a region encoding the extracellular domain, transmembrane domain, or intracellular domain of TMIGD2.

[0068] In some embodiments, the agent comprises a programmable nuclease. In some embodiments, the agent comprises a natural homing meganuclease. In some embodiments, the agent is a TALEN-based agent, a ZFN-based agent, or a CRISPR-based agent, or a biologically active fragment, fusion, derivative, or combination thereof. In some embodiments, the agent is a deaminase or a nucleic acid encoding a deaminase. In some embodiments, the cell is engineered to stably and / or transiently express the TALEN-based agent, the ZFN-based agent, and / or the CRISPR-based agent.

[0069] I. TALEN-based drugs In some embodiments, the gene editing agent is a TALEN-based agent. In some embodiments, the TALEN-based agent is one or more TALEN polypeptides / proteins or biologically active fragments or derivatives thereof, or one or more nucleic acids encoding one or more TALEN polypeptides or fragments or derivatives thereof. Transcription activator-like (TAL) effector sequences can be assembled to specifically bind to target DNA by combining sequences of repeat variable-directions (RVDs). TAL effector and nuclease (TALEN) fusion proteins can make targeted double-strand breaks in cellular DNA, thereby allowing specific genetic modification of cells. In some embodiments, the agent is a TALEN polypeptide / protein or fragment or derivative thereof that targets one or more TMIGD2 DNA sequences. In some embodiments, the RVD portion of the TALEN is engineered to target one or more TMIGD2 DNA sequences. In some embodiments, the TALEN-based agent is a nucleic acid encoding one or more TALEN proteins. In some embodiments, the nucleic acid is present in a plasmid. In some embodiments, the nucleic acid is an mRNA.

[0070] In some embodiments, the TALEN protein is expressed in a cell and induces site-specific double-stranded DNA breaks in one or more TMIGD2 genes. In some embodiments, the TALEN protein introduces a donor sequence that partially or completely replaces the TMIGD2 gene, thereby silencing or inactivating the TMIGD2 gene. In some embodiments, the TALEN is a left TALEN and further comprises a right TALEN that cooperates with it to perform double-stranded breaks in the TMIGD2 gene. In other embodiments, the nucleic acid encoding the TALEN and / or the nucleic acid donor sequence is part of a vector or plasmid. In some embodiments, the TALEN comprises a spacer (e.g., the spacer sequence is 12-30 nucleotides in length).

[0071] Methods for engineering TALENs to bind to specific nucleic acids are described in Cermak, et al, Nucl. Acids Res. 1-1 1 (2011). US Patent Application Publication No. 2011 / 0145940 discloses TAL effectors and methods for using them to modify DNA. Miller et al. Nature Biotechnol 29: 143 (2011) describes the generation of TALENs for site-specific nuclease construction by linking TAL truncation mutants with the catalytic domain of Fok I nuclease. General design principles for TALEN binding domains are described in WO 2011 / 072246. These documents are incorporated herein in their entirety.

[0072] In some embodiments, the TALEN-based agent targets a nucleotide sequence of TMIGD2. In some embodiments, the TALEN-based agent targets a nucleotide sequence that is conserved in multiple strains of TMIGD2. In some embodiments, the TALEN-based agent targets the TMIGD2 pol, env and / or gag genes. In some embodiments, the TALEN-based agent targets the TMIGD2 pol gene. In some embodiments, the TALEN-based agent targets the sequence that encodes the catalytic core of the TMIGD2 pol gene.

[0073] II. ZFN-based drugs In some embodiments, the gene editing agent is a zinc finger nuclease (ZFN)-based agent. In some embodiments, the ZFN-based agent is one or more ZFN polypeptides or biologically active fragments or derivatives thereof, or one or more nucleic acids encoding one or more ZFN polypeptides or fragments or derivatives thereof. ZFNs are artificial restriction enzymes obtained by fusing a zinc finger DNA binding domain with a nuclease. The zinc finger domain can be engineered to target specific desired DNA sequences, allowing zinc finger nucleases to target unique sequences within complex genomes. The DNA binding domain of an individual ZFN typically contains 3-6 zinc finger repeats, each capable of recognizing 9-18 base pairs (bp). If the zinc finger domain perfectly recognizes a 3-base pair DNA sequence to generate a 3-finger array, the array can recognize a 9-base pair target site. In some embodiments, one finger or two finger modules are utilized to generate zinc finger arrays with 6 or more individual zinc fingers. Because the specificity of individual zinc fingers may overlap and depend on the surrounding zinc fingers and DNA context, ZFNs may not be useful for targeting specific TMIGD2.

[0074] A number of selection methods have been developed to generate zinc finger arrays capable of targeting desired sequences. In some embodiments, an initial selection uses phage display to select proteins that bind to specific DNA targets from a large pool of partially randomized zinc finger arrays. In some embodiments, yeast one-hybrid systems, bacterial one-hybrid and two-hybrid systems (e.g., the "OPEN" system), and mammalian cells may be used to select proteins that bind to specific DNA. In particular, the OPEN system combines a preselected pool of individual zinc fingers, each selected to bind to a given triplet, and then uses a second round of selection to obtain a 3-finger array capable of binding to the desired 9 bp sequence.

[0075] In some embodiments, the method of editing the gene sequence of TMIGD2 comprises introducing at least one nucleic acid encoding a zinc finger nuclease capable of recognizing the sequence of TMIGD2 in the genome and cleaving the gene sequence of TMIGD2. In some embodiments, the method further comprises introducing at least one donor polynucleotide comprising a sequence for integration flanked by upstream and downstream sequences that share substantial sequence identity with either side of the cleavage site. In some embodiments, the method further comprises introducing at least one replacement polynucleotide comprising a sequence substantially identical to a portion of the cleavage site of the genomic TMIGD2 sequence and further comprising at least one nucleotide change. In some embodiments, the cultured cells are cultured to express the zinc finger nuclease and introduce a double-stranded break in the genomic TMIGD2 sequence. In some embodiments, the double-stranded break is repaired by a non-homologous end joining repair process, and a silencing or inactivating mutation is introduced into the chromosomal sequence. In some embodiments, the double-stranded break is repaired by a homology-specific repair process, and the sequence in the donor polynucleotide is integrated into the genomic TMIGD2 sequence or the sequence in the replacement polynucleotide is replaced with a portion of the chromosomal sequence.

[0076] In some embodiments, the zinc finger nuclease targets a nucleotide sequence of TMIGD2. In some embodiments, the zinc finger nuclease targets a nucleotide sequence that is conserved across multiple strains of TMIGD2. In some embodiments, the zinc finger nuclease targets the TMIGD2 pol, env and / or gag genes. In some embodiments, the zinc finger nuclease targets the TMIGD2 pol gene. In some embodiments, the zinc finger nuclease targets a sequence encoding the catalytic core of the TMIGD2 pol gene.

[0077] III. CRISPR-based drugs In some embodiments, the gene editing agent is a CRISPR-based agent. In some embodiments, the CRISPR-based agent comprises one or more polynucleotides involved in the expression of CRISPR-associated genes or regulating the activity of CRISPR-associated genes, including but not limited to sequences encoding nuclease genes (e.g., genes encoding Cas9, Cas12a, or Cas13a), tracr (transactivating CRISPR) sequences (e.g., tracrRNA or active portion tracrRNA), tracr-mate sequences (including "direct repeats" and tracrRNA processing portion direct repeats in endogenous CRISPR systems), guide sequences (also called "spacers" in endogenous CRISPR systems), and / or other sequences and transcripts from the CRISPR locus. In some embodiments, the CRISPR-based agent comprises a polynucleotide encoding at least one CRISPR protein and one or more guide RNAs (gRNAs). In some embodiments, the one or more gRNAs comprise sequences that are cognate to PERV polynucleotide sequences and can bind to protospacer adjacent motifs (PAMs). In some embodiments, the PAM comprises the sequence NGG or NNGRRT.

[0078] In some embodiments, the agent is a CRISPR-based polypeptide or a fragment or derivative thereof that targets one or more TMIGD2 DNA sequences. In some embodiments, the CRISPR-based agent is characterized by an element that promotes the formation of a CRISPR complex at the site of the TMIGD2 DNA or RNA sequence. In some embodiments, the CRISPR-based agent is one or more CRISPR / Cas endonucleases or biologically active fragments or derivatives thereof, or one or more nucleic acids encoding one or more CRISPR / Cas polypeptides or fragments or derivatives thereof. In some embodiments, the CRISPR / Cas endonuclease or derivatives thereof is derived from CRISPR type I. In some embodiments, the CRISPR / Cas endonuclease or derivatives thereof is derived from CRISPR type II. In some embodiments, the CRISPR / Cas endonuclease or derivatives thereof is derived from CRISPR type III. In some embodiments, the CRISPR / Cas endonuclease or derivatives thereof is derived from CRISPR type IV. In some embodiments, the CRISPR / Cas endonuclease or derivative thereof is from CRISPR type V. In some embodiments, the CRISPR / Cas endonuclease or derivative thereof is from CRISPR type VI. In some embodiments, the CRISPR / Cas endonuclease or derivative thereof is from CRISPR type IIA, IIB or IIC. In some embodiments, the CRISPR / Cas endonuclease or derivative thereof is from CRISPR type IIC. In some embodiments, the type II CRISPR / Cas endonuclease is Cas9 or a derivative thereof. In some embodiments, the CRISPR / Cas endonuclease or derivative thereof is a type V CRISPR / Cas endonuclease such as Cpf1 (Cas12a) or a derivative thereof. In some embodiments, the CRISPR / Cas endonuclease or derivative thereof is a type VI CRISPR / Cas endonuclease such as Cas13 or a derivative thereof.In some embodiments, the site-directed modifying polypeptide is a type III-B Cmr complex, e.g., a type III-B Cmr complex from Pyrococcus furiosus, Sulfolobus solfataricus, or Thermus thermophilus. See, e.g., Hale, CR et al. Genes & Development, 2014, 28:2432-2443 and Makarova KS et al. Nature Reviews Microbiology, 2015, 13, 1-15.

[0079] In certain embodiments, the CRISPR-based agent utilizes a type II Cas9 endonuclease. In some embodiments, the CRISPR-based agent comprises a type II Cas9 endonuclease and another polynucleotide. In some embodiments, the other polynucleotide is a tracrRNA, a crRNA (also called a "tracr mate RNA") and / or a synthetic single guide RNA (sgRNA). See, e.g., Jinek, M., et al. (2012) Science, 337, 816-821.

[0080] In some embodiments, the CRISPR-based agent is a Cas protein that lacks the ability to cleave double-stranded DNA. In some embodiments, the Cas protein can only cleave a single strand of DNA, i.e., the Cas protein is a "nickase." In some embodiments, the Cas protein cannot cleave either strand of DNA. In some embodiments, the Cas protein is a Cas9 protein that has been mutated to be a nickase or to lack the ability to cleave either strand of DNA. In some embodiments, the Cas9 protein has a D10A and / or H840A mutation. In some embodiments, the agent is a polynucleotide that encodes a Cas9 protein that has a D10A and / or H840A mutation. See, e.g., Cong L., et al. (2013) Science, 339, 819-823; Jinek, M., et al. (2012) Science, 337, 816-821; Gasiunas, G., et al. (2012) Proc. Natl. Acad. Sci. USA, 109, E2579-2586, and Mali, P., et al. (2013) Science, 339, 823-826, which are incorporated by reference in their entireties.

[0081] In some embodiments, the CRISPR-based agent comprises a gRNA. In some embodiments, the gRNA targets a nucleotide sequence of TMIGD2. In some embodiments, the gRNA targets TMIGD2pol, env and / or gag genes. In some embodiments, the gRNA targets TMIGD2pol gene. In some embodiments, the gRNA targets a sequence encoding the catalytic core of TMIGD2pol gene. In some embodiments, the gRNA targets a region that is not the catalytic core of TMIGD2pol gene. In some embodiments, the region that is not the catalytic core of TMIGD2pol gene is upstream of the catalytic core region of TMIGD2pol gene.

[0082] In some embodiments, the agent comprises at least 2 guide RNAs, at least 3 guide RNAs, at least 4 guide RNAs, at least 5 guide RNAs, at least 6 guide RNAs, at least 7 guide RNAs, at least 8 guide RNAs, at least 9 guide RNAs, at least 10 guide RNAs, at least 11 guide RNAs, at least 12 guide RNAs, at least 13 guide RNAs, at least 14 guide RNAs, at least 15 guide RNAs, at least 60 guide RNAs, at least 17 guide RNAs, at least 18 guide RNAs, at least 19 guide RNAs, at least about 20 guide RNAs, at least about 30 guide RNAs, at least about 40 guide RNAs, at least about 50 guide RNAs, at least about 60 guide RNAs, at least about 70 guide RNAs, at least about 80 guide RNAs, at least about 90 guide RNAs, at least about 100 guide RNAs, or more.

[0083] In some embodiments, the CRISPR-based agent is a combination of the CRISPR-based polypeptide / protein and the CRISPR-based polynucleotide disclosed herein. For example, in some embodiments, the CRISPR-based agent comprises a Cas endonuclease and a guide RNA. In some embodiments, the CRISPR-based agent comprises a Cas endonuclease, a tracrRNA, and a tracr mate sequence. In some embodiments, the tracrRNA and the tracr mate sequence are engineered such that they are in the same molecule. In some embodiments, the CRISPR-based agent is one or more polynucleotides encoding any of the above described.

[0084] In some embodiments, the CRISPR-based agent is a chimeric RNA, such as a CRISPR-Cas system RNA. In some embodiments, the CRISPR-based agent has at least one second guide sequence that can hybridize to a nucleic acid molecule expressing an RNA sequence of the CRISPR-Cas system or a component of a CRISPR-Cas complex to reduce or eliminate functional expression of the system or complex, which can be self-inactivated, and the second guide sequence can hybridize to a nucleic acid molecule for expression of a CRISPR enzyme.

[0085] In some aspects, the disclosure provides methods of using the CRISPR-based agents disclosed herein. In some aspects, the disclosure provides effective means of modifying TMIGD2 polynucleotide sequences using the CRISPR-based agents disclosed herein. The CRISPR complexes of the invention have a wide variety of utilities, including modifying (e.g., deleting, inactivating) TMIGD2 polynucleotide sequences in a variety of cells from a variety of tissues and organs. Thus, the CRISPR complexes of the invention have a wide range of applications, for example, in gene or genome editing.

[0086] In some aspects, the disclosure provides a method of genome editing in vivo, comprising providing a quantity of one or more vectors, each encoding at least one CRISPR protein and one or more guide RNAs (gRNAs), and administering the one or more vectors to a mammal, wherein in vivo expression of the one or more vectors comprises binding of the CRISPR protein to the TMIGD2 locus cognate to the gRNA and in vivo generation of a double strand break (DSB) in a cell population in the mammal, and in vivo homologous recombination (HR) of the DSB results in genome editing of the cell population in the mammal. In some aspects, the CRISPR protein is Cas9, and the one or more gRNAs comprise a sequence capable of binding to a protospacer adjacent motif ("PAM"). In some aspects, the HR comprises non-homologous end joining (NHEJ) to introduce a missense or nonsense in a protein expressed at the PERV locus.

[0087] In some embodiments, the CRISPR-based agent further comprises a moiety that regulates expression of TMIGD2. In some embodiments, the CRISPR-based agent is a fusion protein that includes a transcriptional repressor domain.

[0088] E. Combination Therapy In some embodiments, the agent that inhibits TMIGD2 expression, activity, or both is administered with at least a second or additional agent. In certain of these embodiments, the agent that inhibits TMIGD2 expression, activity, or both is administered in combination with an immune checkpoint inhibitor. Immune checkpoint proteins are well known in the art and include, but are not limited to, CTLA-4, PD-1, VISTA, B7-H2, B7-H3, PD-L1, B7-H4, B7-H6, 2B4, ICOS, HVEM, PD-L2, CD160, gp49B, PIR-B, KIR receptor family, TIM-1, TIM-3, TIM-4, LAG-3, BTLA, SIRPα (CD47), CD48, 2B4 (CD244), B7.1, B7.2, ILT-2, ILT-4, TIGIT, HHLA2, KIR3DL3, and A2aR. (See, e.g., International Publication No. WO 2012 / 177624, incorporated herein by reference in its entirety.) Inhibition of one or more immune checkpoint inhibitors can block or neutralize inhibitory signaling, thereby upregulating the immune response to more effectively treat cancer.

[0089] In certain embodiments, the agent that inhibits TMIGD2 expression, activity, or both is administered in combination with, for example, chemotherapeutic agents, hormones, anti-angiogenic agents, radiolabeled compounds, or surgery, cryotherapy, and / or radiation therapy. The aforementioned therapies can be administered in combination with other forms of existing therapy (e.g., standard of care cancer treatments well known to those of skill in the art), either prior to or subsequent to the existing therapy. For example, the agents described herein can be administered with a therapeutically effective amount of a chemotherapeutic agent. Such chemotherapeutic agents can include, but are not limited to, platinum compounds, cytotoxic antibiotics, antimetabolites, antimitotic agents, alkylating agents, arsenic compounds, DNA topoisomerase inhibitors, taxanes, nucleoside analogs, plant alkaloids, and toxins and their synthetic derivatives. Representative compounds include, but are not limited to, alkylating agents: cisplatin, treosulfan, and trofosfamide, plant alkaloids: vinblastine, paclitaxel, docetaxol, DNA topoisomerase inhibitors: teniposide, crisnatol, and mitomycin, antifolates: methotrexate, mycophenolic acid, and hydroxyurea, pyrimidine analogs: 5-fluorouracil, doxifluridine, and cytosine arabinoside, purine analogs: mercaptopurine and thioguanine, DNA antimetabolites: 2'-deoxy-5-fluorouridine, aphidicolin glycinate, and pyrazoloimidazole, antimitotics: halichondrin, colchicine, and rhizoxin. Compositions containing one or more chemotherapeutic agents (e.g., FLAG or CHOP) may be used. FLAG comprises fludarabine, cytosine arabinoside (Ara-C), and G-CSF. CHOP comprises cyclophosphamide, vincristine, doxorubicin and prednisone. The foregoing examples of chemotherapy drugs are illustrative and not intended to be limiting.

[0090] In another embodiment, the agent that inhibits TMIGD2 expression, activity, or both is administered in combination with radiation therapy. The radiation used in radiation therapy may be ionizing radiation. Radiation therapy may be gamma radiation, X-rays, or protons. Examples of radiation therapy include, but are not limited to, external beam radiation therapy, tissue implantation of radioisotopes, radioisotopes such as strontium-89 (I125, palladium, iridium), chest radiation therapy, intraperitoneal P32 radiation therapy, and / or whole abdominal and pelvic radiation therapy. For a general overview of radiation therapy, see Hellman, Chapter 16: Principles of Cancer Management: Radiation Therapy, 6th edition, 2001, DeVita et al., eds., JB Lippencott Company, Philadelphia. Radiation therapy can be performed as external beam radiation or teletherapy, where radiation is delivered from a remote source. Radiation therapy can also be performed as internal therapy or brachytherapy, where a radiation source is placed inside the body near the cancer cells or tumor mass. Also included are the use of photosensitizers such as hematoporphyrin and its derivatives, vertoporfm (BPD-MA), phthalocyanines, photosensitizer Pc4, demethoxyhypocrelin A, and photodynamic therapy, including administration of 2B A-2-DMHA.

[0091] In some embodiments, the agent that inhibits TMIGD2 expression, activity, or both is administered in combination with hyperthermia, photodynamic therapy, and / or surgery. In such embodiments, the hyperthermia treatment is or includes local hyperthermia (e.g., external, intracavitary, or interstitial hyperthermia), local hyperthermia (e.g., deep tissue hyperthermia), local perfusion (e.g., continuous hyperthermic peritoneal irrigation), or whole-body hyperthermia. In some embodiments, the photodynamic therapy is or includes the administration of a photosensitizer, such as hematoporphyrin and its derivatives, verteporfin (BPD-MA), phthalocyanines, photosensitizer Pc4, demethoxyhypocrelin A, 2BA-2-DMHA, or combinations thereof. In some embodiments, the surgery is or includes surgery to remove cancerous or precancerous tissue. In some embodiments, the transplant is or includes stem cell transplant or organ transplant.

[0092] In another embodiment, an agent that inhibits TMIGD2 expression, activity, or both is administered in combination with hormone therapy, which can include, for example, hormone agonists, hormone antagonists (e.g., flutamide, bicalutamide, tamoxifen, raloxifene, leuprolide acetate (leupron), LH-RH antagonists), inhibitors of hormone biosynthesis and processing, and steroids (e.g., dexamethasone, retinoids, deltoids, betamethasone, cortisol, cortisone, prednisone, dehydrotestosterone, glucocorticoids, mineralocorticoids, estrogens, testosterone, progestins), vitamin A derivatives (e.g., all-trans retinoic acid (ATRA)); vitamin D3 analogs; antigestagens (e.g., mifepristone, onapristone), or antiandrogens (e.g., cyproterone acetate).

[0093] In one embodiment, the agent that inhibits TMIGD2 expression, activity, or both is administered in combination with an immunomodulatory interleukin, such as IL-2, IL-6, IL-7, IL-12, IL-17, IL-23, and a modulator thereof (e.g., a blocking antibody, or a more potent or longer lasting form). In another embodiment, the agent that inhibits TMIGD2 expression, activity, or both is administered in combination with an immunomodulatory cytokine, such as interferon, G-CSF, imiquimod, TNFα, and a modulator thereof (e.g., a blocking antibody, or a more potent or longer lasting form). In another embodiment, the agent that inhibits TMIGD2 expression, activity, or both is administered in combination with an immunomodulatory chemokine, such as CCL3, CCL26, and CXCL7, and a modulator thereof (e.g., a blocking antibody, or a more potent or longer lasting form). In another embodiment, the agent that inhibits TMIGD2 expression, activity, or both is administered in combination with an immunomodulatory molecule that targets immunosuppression, such as a STAT3 signaling modulator, an NFκB signaling modulator.

[0094] In certain aspects, the agent that inhibits TMIGD2 expression, activity or both is an immune cytostatic, a glucocorticoid, a cytostatic, an immunophilin and its modulators (e.g., rapamycin, calcineurin inhibitors, tacrolimus, cyclosporine, pimecrolimus, avetimus, gusperimus, ridaforolimus, everolimus, temsirolimus, zotarolimus, etc.), hydrocortisone (cortisol), cortisone acetate, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, triamcinolone, beclomethasone, fludrocortisone acetate, deoxycorticosterone acetate (doca), aldosterone, a non-glucocorticoid steroid, a pyrimidine synthesis inhibitor, leflunomide, teriflunomide, a folic acid analog, methotrexate, antithymocyte globulin, an anti Lymphocyte globulin, thalidomide, lenalidomide, pentoxifylline, bupropion, curcumin, catechin, opioids, IMPDH inhibitors, mycophenolic acid, myriocin, fingolimod, NF-xB inhibitors, raloxifene, drotrecogin alfa, denosumab, NF-xB signaling cascade inhibitors, disulfiram, olmesartan, dithiocarbamates, proteasome inhibitors, bortezomib, MG132, Pro1, NPI-0052, curcumin, genistein, resveratrol, parthenolide, thalidomide, lenalidomide, flavopiridol, nonsteroidal anti-inflammatory drugs (NSAIDs), arsenic trioxide, dehydroxymethylepoxyquinomycin (DHMEQ), I3C (indole-3-carbinol) / DIM (diindolemethane) (13C / DIM), Bay Administered in combination with immunomodulatory agents such as 11-7082, luteolin, the membrane-permeable peptide SN-50, overexpression of IKBa-super-repressor, NFκB decoy oligodeoxynucleotides (ODN) or derivatives or analogs thereof.

[0095] In one embodiment, the agent that inhibits TMIGD2 expression, activity, or both is an antibody that binds to CD40, Toll-like receptors (TLRs), OX40, GITR, CD27, or 4-1BB, a T cell bispecific antibody, an anti-IL-2 receptor antibody, an anti-CD3 antibody, OKT3 (muromonab), otelixizumab, teplizumab, visilizumab, an anti-CD4 antibody, clenoliximab, keliximab, zanolimumab, efalizumab, an anti-CD18 antibody, erlizumab, rovelizumab, an anti-CD20 antibody, afutuzumab, ocrelizumab, ofatumumab, pascolizumab, rituximab, an anti-CD23 antibody, rumiliximab, an anti-CD40 antibody, teneliximab, toralizumab, an anti-CD40L antibody, rupriximab ... izumab, anti-CD62L antibody, acelizumab, anti-CD80 antibody, galiximab, anti-CD147 antibody, gavilimomab, B-lymphocyte stimulator (BLyS) inhibitor antibody, belimumab, CTLA4-Ig fusion protein, abatacept, belatacept, anti-CTLA4 antibody, ipilimumab, tremelimumab, anti-eotaxin 1 antibody, bertilimumab, anti-α4 integrin antibody, natalizumab, anti-IL-6R antibody, tocilizumab, anti-LFA-1 antibody, odulimomab, anti-CD25 antibody, basiliximab, daclizumab, inolimomab, anti-CD5 antibody, zolimomab, anti-CD2 antibody, siplizumab, nerelimomab, faralimomab, atlizumab, atolimumab, cedelizumab, dorlimomab Administered in combination with immune-modulating antibodies or proteins such as alitox, dorlixizumab, fontolizumab, gantenerumab, gomilikimab, lebrilizumab, maslimomab, morolimumab, pexelizumab, reslizumab, rovelizumab, talizumab, terimomab alitox, bapaliximab, beparimomab, aflibercept, alefacept, rilonacept, IL-1 receptor antagonists, anakinra, anti-IL-5 antibodies, mepolizumab, IgE inhibitors, omalizumab, talizumab, IL12 inhibitors, IL23 inhibitors, and ustekinumab.

[0096] In some embodiments, an agent that inhibits TMIGD2 expression, activity, or both is administered in combination with adoptive cell-based immunotherapy, including, but not limited to, irradiated autologous or allogeneic tumor cells, tumor lysates or apoptotic tumor cells, antigen-presenting cell-based immunotherapy, dendritic cell-based immunotherapy, adoptive T cell transfer, adoptive CAR T cell therapy, natural killer (NK) cells, autoimmune enhancement therapy (AIET), cancer vaccines, antigen-presenting cells, and / or combinations thereof. Such cell-based immunotherapies can be further modified to express one or more gene products to further modulate the immune response, such as to express cytokines such as GM-CSF, and / or to express tumor-associated antigens (TAA), such as Mage-1, gp-100, patient-specific neo-antigen vaccines, etc.

[0097] Manufacturing method This disclosure provides, inter alia, methods for producing the antibodies or antigen-binding fragments thereof described herein. In some embodiments, the antibodies or antigen-binding fragments thereof described herein are identified using display technologies such as yeast display, phage display, or ribosome display. In some embodiments, the antibodies or antigen-binding fragments thereof described herein are identified in a hybridoma library (e.g., a mammalian hybridoma library, such as a murine hybridoma library), followed by screening of the supernatants.

[0098] Combinatorial methods for generating antibodies or antigen-binding fragments thereof have been described in (e.g., U.S. Pat. No. 5,223,409 to Ladner et al.; WO 92 / 18619 to Kang et al.; WO 91 / 17271 to Dower et al.; WO 92 / 20791 to Winter et al.; WO 92 / 15679 to Markland et al.; WO 93 / 01288 to Breitling et al.; WO 92 / 01047 to McCafferty et al.; WO 92 / 09690 to Garrard et al.; WO 90 / 02809 to Ladner et al.; Fuchs et al. (1991) Bio / Technology 9:1370-1372; Hay et al. (1992) Hum Antibody Hybridomas 3:81-85;Huse et al. (1989) Science 246:1275-1281;Griffths et al. (1993) EMBO J 12:725-734;Hawkins et al. (1992) J Mol Biol 226:889-896;Clackson et al. (1991) Nature 352:624-628; Gram et al. (1992) PNAS 89:3576-3580; Garrad et al. (1991) Bio / Technology 9:1373-1377; Hoogenboom et al. (1991) Nuc Acid Res 19:4133-4137 and Barbas et al. (1991) PNAS 88:7978-7982, which are incorporated herein by reference in their entireties.

[0099] The antibodies or antigen-binding fragments thereof described herein may be derived from other species. Humanized antibodies are antibodies produced by recombinant DNA techniques in which some or all of the amino acids of a human immunoglobulin light or heavy chain that are not necessary for binding to an antigen (e.g., constant and / or framework regions of the variable domain) are used to replace the corresponding amino acids of the light or heavy chain of a cognate non-human antibody. As an example, a humanized version of a mouse antibody against an antigen would have in its heavy and light chains: (1) the constant regions of a human antibody; (2) the FRs of the variable domain of a human antibody; and (3) the CDRs of a mouse antibody. The human FRs may be selected based on the highest sequence homology to the mouse FR sequences. If necessary, one or more residues of the human FRs can be changed to the residues at the corresponding positions in the mouse antibody so as to maintain the binding affinity of the humanized antibody with its target. This change is sometimes called a "back mutation." Similarly, forward mutations may be used to return to the mouse sequence for desired reasons, such as stability or affinity with the target. Because humanized antibodies have significantly fewer non-human components, they are generally less likely to provoke an immune response in humans than chimeric human antibodies.

[0100] Methods for humanizing non-human antibodies are widely known in the art. Suitable methods for producing humanized antibodies according to this disclosure are described, for example, in Winter's European Patent Application Publication No. 0 239 400; Jones et al., Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-327 (1988); Verhoeyen et al., Science 239: 1534-1536 (1988); Queen et al., Proc. Nat. Acad. ScL USA 86:10029 (1989); U.S. Patent No. 6,180,370 and Orlandi et al., Proc. Natl. Acad. Sd. USA 86:3833 (1989) (the disclosures of which are incorporated herein by reference in their entirety). In general, grafting of non-human (e.g., murine) CDRs onto a human antibody is performed as follows: VH- and VL-encoding cDNAs are isolated from hybridomas, and the sequences of the nucleic acids encoding the VH and VL containing the CDRs are determined. The sequences of the nucleic acids encoding the CDRs are inserted into the corresponding regions of a human antibody VH or VL coding sequence and combined with human constant region gene segments of the desired isotype (e.g., γ1 for CH, κ for CL). The humanized heavy and light chain genes are co-expressed in mammalian host cells (e.g., CHO or NSO cells) to produce soluble humanized antibodies. To facilitate large-scale production of antibodies, it is often desirable to select for highly expressing genes, for example, using the DHFR or GS genes of the production strain.

[0101] The antibodies or antigen-binding fragments thereof described herein may be or may include human antibodies or antigen-binding fragments thereof. Fully human antibodies may be particularly desirable for treating human subjects. Human antibodies can be made by a variety of methods known in the art including the above-mentioned phage display methods using antibody libraries derived from human immunoglobulin sequences (see, e.g., U.S. Pat. Nos. 4,444,887 and 4,716,111; and WO 98 / 46645, 98 / 60433, 98 / 24893, 98 / 16664, 96 / 34096, 96 / 33735, and 91 / 10741, which are incorporated by reference in their entireties.) For example, techniques for producing human monoclonal antibodies can be used as described in Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Riss, (1985) and Boerner et al., J. Immunol., 147(1):86-95, (1991), which are incorporated by reference in their entireties.

[0102] The antibodies or antigen-binding fragments thereof described herein may be or include chimeric antibodies or antigen-binding fragments thereof. Methods for describing chimeric antibodies are described, for example, in U.S. Patent No. 4,816,567 and Morrison et al., Proc. Natl. Acad. Sci. USA, 1984, 81:6851-6855, which are incorporated by reference in their entirety. In some embodiments, chimeric antibodies are produced by recombinant techniques that combine non-human variable regions (e.g., variable regions derived from a non-human primate such as a mouse, rat, hamster, rabbit, or monkey) with human constant regions.

[0103] Suitable methods can be used to introduce mutations into one or more polynucleotide sequences encoding the antibodies or antigen-binding fragments thereof described herein, including error-prone PCR, chain shuffling, and oligonucleotide-directed mutagenesis such as trinucleotide-directed mutagenesis (TRIM). In some embodiments, several CDR residues (e.g., 4-6 residues at a time) are randomized. CDR residues involved in antigen binding may be specifically identified, for example, by alanine scanning mutagenesis or modeling. CDR-H3 and CDR-L3 in particular are often targeted for mutations. A secondary library can be produced by introducing mutations into the variable regions and / or CDRs. The secondary library is then screened to identify antibody variants with improved affinity. Construction of secondary libraries and affinity maturation by reselection is described, for example, in Hoogenboom et al., Methods in Molecular Biology, 2001, 178:1-37, which is incorporated by reference in its entirety.

[0104] composition Compositions comprising one or more of the agents used in the methods provided herein, as well as the use of these agents to inhibit the expression and / or activity of TMIGD2, are provided in certain aspects herein.

[0105] In some embodiments, the compositions provided herein are pharmaceutical compositions comprising one or more agents that inhibit the expression and / or activity of TMIGD2 and a pharma- ceutically acceptable excipient. Non-limiting examples of pharma-ceutically acceptable excipients include those described in, for example, "Remington: The Science and Practice of Pharmacy", 19th Ed. (1995) Mack Publishing Co or its latest edition; A. Gennaro (2000) "Remington: The Science and Practice of Pharmacy", 20th edition, Lippincott, Williams, & Wilkins; Pharmaceutical Dosage Forms and Drug Delivery Systems (1999) HC Ansel et al., eds., 7th ed., Lippincott, Williams, & Wilkins, and Handbook of Pharmaceutical Excipients (2000) AH Kibbe et al., eds., 3rd ed. Amer. Pharmaceutical Assoc. In some embodiments, the compositions are suitable for administration to a subject, e.g., are sterile compositions. In some embodiments, the compositions are suitable for administration to a human subject, e.g., the compositions are sterile and free of detectable pyrogens and / or other toxins.

[0106] In some embodiments, the compositions include other ingredients such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, talc, cellulose, glucose, sucrose, magnesium, carbonates, etc. In some embodiments, the compositions include pharma- ceutically acceptable auxiliary substances as needed to approximate physiological conditions, such as pH adjusters and buffers, toxicity adjusters, etc., e.g., sodium acetate, sodium chloride, potassium chloride, calcium chloride, sodium lactate, hydrochloride salts, sulfate salts, solvates (e.g., mixed ionic salts, water, organics), hydrates (e.g., water), etc.

[0107] In some embodiments, the composition is in the form of an aqueous solution, powder, granules, tablets, pills, suppositories, capsules, suspensions, sprays, etc. The composition may contain pharma- ceutically acceptable additives, pharma-ceutically acceptable salts, diluents, carriers, vehicles, and other inert agents well known to those skilled in the art. Vehicles and additives commonly used in pharmaceutical preparations include, for example, talc, gum arabic, lactose, starch, magnesium stearate, cocoa butter, aqueous or non-aqueous solvents, oils, paraffin derivatives, glycols, and the like. Solutions can be prepared using water or physiologically compatible organic solvents such as ethanol, 1,2-propylene glycol, polyglycols, dimethyl sulfoxide, fatty alcohols, triglycerides, partial esters of glycerin, and the like. Parenteral compositions can be prepared using conventional techniques that may include sterile isotonic saline, water, 1,3-butanediol, ethanol, 1,2-propylene glycol, polyglycols mixed with water, Ringer's solution, and the like. In one aspect, a coloring agent is added to facilitate finding the composition and properly placing it at the intended treatment site.

[0108] The composition may contain preservatives and / or stabilizers. Non-limiting examples of preservatives include methylparaben, ethylparaben, propylparaben, sodium benzoate, benzoic acid, sorbic acid, potassium sorbate, propionic acid, benzalkonium chloride, benzyl alcohol, thimerosal, phenylmercuric salts, chlorhexidine, phenol, 3-cresol, quaternary ammonium compounds (QACs), chlorobutanol, 2-ethoxyethanol, and imidurea.

[0109] To control tonicity, the composition may include a physiological salt such as a sodium salt. Sodium chloride (NaCl) is preferred and may be present at 1-20 mg / ml. Other salts that may be present include potassium chloride, potassium dihydrogen phosphate, disodium hydrogen phosphate dihydrate, magnesium chloride and calcium chloride.

[0110] The composition may contain one or more buffers. Representative buffers include phosphate buffer, Tris buffer, borate buffer, succinate buffer, histidine buffer or citrate buffer. Buffers are usually present at a concentration of 5 to 20 mM. The pH of the composition is generally 5 to 8, more typically 6 to 8, for example 6.5 to 7.5 or 7.0 to 7.8.

[0111] The compositions may be administered by any suitable route apparent to one of skill in the art depending on the disease or condition being treated. Exemplary routes of administration include intravenous, intraarterial, intramuscular, subcutaneous, intracranial, intranasal or intraperitoneal.

[0112] In some embodiments, the composition may include a cryoprotectant, non-limiting examples of which include glycols (e.g., ethylene glycol, propylene glycol, and glycerin), dimethyl sulfoxide (DMSO), formamide, sucrose, trehalose, dextrose, and combinations thereof.

[0113] The composition may include pharma- ceutically acceptable additives, pharma- ceutically acceptable salts, diluents, carriers, vehicles, and other inert agents well known to those skilled in the art. Vehicles and additives commonly used in pharmaceutical preparations include, for example, talc, gum arabic, lactose, starch, magnesium stearate, cocoa butter, aqueous or non-aqueous solvents, oils, paraffin derivatives, glycols, and the like. Solutions may be prepared using water or physiologically compatible organic solvents, such as ethanol, 1,2-propylene glycol, polyglycols, dimethyl sulfoxide, fatty alcohols, triglycerides, partial esters of glycerin, and the like. Parenteral compositions may be prepared using conventional techniques, which may include sterile isotonic saline, water, 1,3-butanediol, ethanol, 1,2-propylene glycol, polyglycols mixed with water, Ringer's solution, and the like. In some aspects, coloring agents are added to facilitate locating the composition and properly placing it at the intended treatment site.

[0114] As can be seen from the disclosure above, the present invention has a variety of applications. The present invention is further described in the following examples, which are merely illustrative and are not intended to limit the definition and scope of the invention in any way. EXAMPLES

[0115] Targeting TMIGD2 to treat hematological malignancies The following examples show that TMIGD2 is expressed in various human hematological malignancies, is functionally important for leukemia-initiating cells, and is associated with poor overall survival in AML patients. The examples also show that knockdown of TMIGD2 reduces the maintenance of AML stem cells and increases cell death in human hematological malignancies. Furthermore, the examples show that treatment with anti-TMIGD2 monoclonal antibodies inhibits AML progression in vivo. Taken together, the results of this example suggest that targeting the expression of TMIGD2 can be used as a therapeutic approach for hematological malignancies.

[0116] TMIGD2 is highly expressed in a variety of hematological malignancies Although TMIGD2 has been identified as a member of the CD28 family and a receptor for HHLA2, the expression of TMIGD2 at the protein level in human tumor cells remained unclear. To investigate the protein expression, we used fluorescence-activated cell sorting (FACS) and monoclonal antibody (mAb) against TMIGD2 to test the TMIGD2 protein on various hematological malignancies. The results showed that the expression of TMIGD2 protein on the cell surface of three tumor lines, human erythroleukemia (HEL), chronic myeloid leukemia (K562), and acute myeloid leukemia (Kg1a), was at a high level (Figure 1A). TMIGD2 mRNA was highly expressed in cell lines, including human leukemia, lymphoma, and multiple myeloma (Figure 1B).

[0117] TMIGD2, but not PD-L1 / PD-1, mRNA is highly expressed in human acute myeloid leukemia (AML) and is associated with poorer overall survival in patients The Cancer Genome Atlas (TCGA) and Genotype-Tissue Expression (GTEx) datasets included 173 human acute myeloid leukemia (AML) and 70 normal bone marrow samples, and the following analyses included investigating the mRNA expression of the HHLA2 / TMIGD2 / KIR3DL3 and PDL1 / PD-1 pathways.

[0118] Analysis revealed that TMIGD2 mRNA levels were significantly higher in AML samples than in normal bone marrow (Figure 2A), HHLA2 expression was also higher in AML samples but not statistically significant (Figure 2A), and KIR3DL3 expression was very low. In contrast to TMIGD2, PD-1 mRNA levels were significantly lower in AML samples than in normal bone marrow samples (Figure 2B).

[0119] AML samples were further divided into two groups according to the expression level of TMIGD2: TMIGD2-high group (top 25%) and TMIGD2-low group (remaining 75%). The results showed that the overall survival rate of the TMIGD2-high group was significantly (p=0.011) lower than that of the TMIGD2-low group (Figure 2C). Collectively, these results suggest that TMIGD2, but not the traditionally known pathway of PD-L1 / PD-1, is highly expressed in human AML and is associated with worse overall survival of patients.

[0120] TMIGD2 is highly expressed in AML stem / progenitor cells The expression of TMIGD2 protein in peripheral blood cells from 40 AML patients, cord blood mononuclear cells from five healthy donors, and bone marrow cells from five healthy adults was measured by FACS. The experiment results showed that TMIGD2-positive cells were significantly more abundant in CD34+ stem / progenitor cells than in CD34- differentiated blast cells from AML patients (Fig. 3A, P<0.0001). Furthermore, TMIGD2-positive cells in CD34+ stem / progenitor cells from AML patients were significantly more abundant than normal CD34+ stem / progenitor cells in cord blood / bone marrow mononuclear cells from healthy donors (Fig. 3B, P<0.01).

[0121] TMIGD2 enriches functional leukemia-initiating cells TMIGD2 is overexpressed in AML stem cells (Figures 3A-3B), and thus TMIGD2+ and TMIGD2- AML stem cells (CD45 dim SSC low Two sets of experiments were performed to directly compare the frequency of leukemia-initiating cells between Lin- (CD3-CD14-CD19-) and CD34+CD38-.

[0122] First, TMIGD2+ and TMIGD2- AML stem cells from AML samples were sorted by FACS (Figure 4A), and then the purified cells were seeded in methylcellulose-based medium for in vitro colony-forming unit (CFU) assay. The formed colonies were counted and classified based on their unique morphology. The colony cells were collected and replated to examine their self-renewal ability. Compared with TMIGD2- AML stem cells from the same patient, TMIGD2+ AML stem cells formed colonies with much higher CFU numbers after 14 days of culture in both the first and second cultures (Figure 4B).

[0123] Second, we performed in vivo limiting dilution xenotransplantation experiments by FACS sorting TMIGD2+ and TMIGD2- AML stem cells from the same AML samples and then transplanting the two sorted populations into sublethally irradiated NSG mice. After 12 weeks or more, bone marrow cells from these NSG mice were analyzed by FACS to measure lymphoid and myeloid engraftment.

[0124] The frequencies of leukemia-initiating cells between TMIGD2+ and TMIGD2- AML stem cells from the same patient (patient #31) were found to be 1 / 399 and 1 / 10985, respectively (Figure 4C). These data indicate that TMIGD2 enriches for functional leukemia-initiating cells.

[0125] RNA-seq comparison between CD34+TMIGD2+ and CD34+TMIGD2- populations demonstrated that TMIGD2+ AML stem cells were associated with established leukemia stem cells (LSCs) and a 17-gene stemness signature (Figure 4D).

[0126] Knockdown of TMIGD2 reduces maintenance of AML stem cells TMIGD2 is overexpressed in AML stem cells (Figures 3A-3B) and associated with worse overall patient survival (Figures 2A-2C), suggesting that TMIGD2 plays an important role in AML stem cells. To analyze the function of TMIGD2, TMIGD2+ AML stem cells (CD45dimSSClowLin- (CD3-CD14-CD19-) CD34+ CD38-) were sorted from AML peripheral blood by FACS, transduced with lentivirus expressing scrambled control shRNA (shCtrl) or TMIGD2-specific shRNA (shTMIGD2) (Figure 5A), and sorted by GFP 3 days after transduction. As shown in Figure 5B, compared with shCtrl, shTMIGD2 largely reduced the expression of TMIGD2 in AML stem cells. We then performed a CFU assay and found that knockdown of TMIGD2 in AML stem cells significantly reduced colony formation in all three AML patient samples (Figure 5C). These results indicate that TMIGD2 is functionally important for the maintenance of AML stem cells and that targeting TMIGD2 reduces AML stem cell survival.

[0127] Knockdown of TMIGD2 increases cell death in human hematological malignancies To investigate the role of TMIGD2 in AML, we knocked down TMIGD2 in HEL cells using lentivirus-mediated shRNA. We found that knockdown of TMIGD2 enhanced both early apoptosis (Annexin V+DAPI-) and late apoptosis / necrosis (Annexin V+DAPI+) in HEL cells (Figure 6A). To understand the molecular mechanism by which TMIGD2 regulates AML functions, we analyzed RNA-seq data from HEL-shCtrl and HEL-shTMIGD2 cells. The analysis revealed that shTMIGD2 knockdown HEL cells were significantly enriched for genes involved in apoptosis and cell cycle arrest compared to shCtrl cells (Figure 6B). These findings support that TMIGD2 is required for AML cell survival and proliferation.

[0128] Treatment with anti-TMIGD2 monoclonal antibody suppresses AML progression in vivo To investigate the therapeutic effect of anti-TMIGD2 mAbs in AML in vivo, NSG mice were fed with patient-derived AML cells and then treated with anti-TMIGD2 mAbs 20F2 and 17C7. We found that anti-TMIGD2 mAbs suppressed AML progression in vivo (Figure 5D). These findings support that mAbs against TMIGD2 can be used to treat AML.

[0129] References 1. Janakiram, M., et al. The third group of the B7-CD28 immune checkpoint family: HHLA2, TMIGD2, B7x, and B7-H3. Immunol Rev 276, 26-39 (2017). 2. Zang, X. & Allison, JP The B7 family and cancer therapy: costimulation and coinhibition. Clin Cancer Res 13, 5271-5279 (2007). 3. John, P., et al. The B7x immune checkpoint pathway: From discovery to clinical trial. Trends Pharmacol Sci 40, 883-896 (2019). 4. Ohaegbulam, KC, Assal, A., Lazar-Molnar, E., Yao, Y. & Zang, X. Human cancer immunotherapy with antibodies to the PD-1 and PD-L1 pathway. Trends Mol Med 21, 24-33 (2015). 5. Chinai, J.M., et al. New immunotherapies targeting the PD-1 pathway. Trends Pharmacol Sci 36, 587-595 (2015). 6. Vincenti, F., Dritselis, A. & Kirkpatrick, P. Belatacept. Nat Rev Drug Discov 10, 655-656 (2011). 7. Zhao, R., et al. HHLA2 is a member of the B7 family and inhibits human CD4 and CD8 T-cell function. Proc Natl Acad Sci U S A 110, 9879-9884 (2013). 8. Janakiram, M., Chinai, J.M., Zhao, A., Sparano, J.A. & Zang, X. HHLA2 and TMIGD2: new immunotherapeutic targets of the B7 and CD28 families. Oncoimmunology 4, e1026534 (2015). 9. Janakiram, M., et al. Expression, clinical significance, and receptor identification of the newest B7 family member HHLA2 protein. Clin Cancer Res 21, 2359-2366 (2015). 10. Zhuang, X. & Long, E.O. CD28 Homolog Is a Strong Activator of Natural Killer Cells for Lysis of B7H7(+) Tumor Cells. Cancer Immunol Res 7, 939-951 (2019). 11. Zhu, Y., et al. B7-H5 costimulates human T cells via CD28H. Nat Commun 4, 2043 (2013). 12. Wei, Y., et al. KIR3DL3-HHLA2 is a human immunosuppressive pathway and a cancer therapeutic target. Sci Immunol (2021). in Reversion 13. Zang, X. New immune checkpoint pathways: HHLA2 and its receptors including TMIGD2. Cold Spring Harbor Asia Conference on Precision Cancer Biology: From targeted immune therapies (2017).

Claims

1. Use of an agent that inhibits the expression, activity or both of TMIGD2 in the manufacture of a medicament for preventing or treating cancer in a subject in need thereof, said use comprising administering an effective amount of said agent to said subject.

2. The use according to claim 1, wherein said agent is selected from the group consisting of an antibody agent, an mRNA targeting agent, a small molecule agent and a gene editing agent.

3. The use according to claim 2, wherein said antibody agent is an antibody or an antigen-binding fragment thereof that specifically binds to an epitope within the extracellular domain of TMIGD2.

4. The use according to claim 3, wherein the extracellular domain of said TMIGD2 comprises residues 1 to 150 of the amino acid sequence shown in SEQ ID NO: 1 or SEQ ID NO: 2, or residues 1 to 30 of the amino acid sequence shown in SEQ ID NO:

3.

5. Said antibody or antigen-binding fragment thereof (a) has a heavy chain variable region comprising GYTFTSYDIN (SEQ ID NO: 24), WIYPGDGSTNYNEKFKG (SEQ ID NO: 25), and ARRLRYYFDY (SEQ ID NO: 26) ; and / or a light chain variable region comprising RASQDIRNYLN (SEQ ID NO: 32), YTSRLHS (SEQ ID NO: 33), and QQVNTPWT (SEQ ID NO: 34) ; or (b) has a heavy chain variable region comprising GYSITSDYAWN (SEQ ID NO: 56), YITYSGSSTSYNPSLKS (SEQ ID NO: 57), and ARSGYRYDDAMDY (SEQ ID NO: 58) ; and / or a light chain variable region comprising KSSQSLSSNNQKNYLA (SEQ ID NO: 64), FASTRES (SEQ ID NO: 65), and QQHYRTPLT (SEQ ID NO: 66) ; The use according to claim 3, comprising.

6. Said antibody or antigen-binding fragment thereof (a) has a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 23; and / or a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 31; or (b) has a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 55; and / or a light chain variable region comprising the amino acid sequence shown in SEQ ID NO:

63. The use according to claim 5, comprising.

7. The use according to claim 2, wherein said mRNA targeting agent is an antisense agent or an RNAi agent.

8. The antisense agent comprises, or consists of, a nucleic acid sequence complementary to the mRNA encoded by the nucleic acid sequence shown in SEQ ID NO: 4, SEQ ID NO: 5 or SEQ ID NO: 6, or the antisense agent comprises, or consists of, a nucleic acid sequence having a sequence identity of about 80% or more with respect to the mRNA encoded by the nucleic acid sequence shown in SEQ ID NO: 4, SEQ ID NO: 5 or SEQ ID NO:

6. The use according to claim 7.

9. The RNAi agent is selected from the group consisting of small interfering RNA (siRNA), double-stranded RNA (dsRNA), microRNA (miRNA), piwiRNA (piRNA), small nucleolar RNA (snoRNA), tRNA-derived small RNA (tsRNA), small regulatory RNA (srRNA), and small hairpin RNA (shRNA) molecules. The use according to claim 7.

10. The RNAi agent comprises, or consists of, a nucleic acid sequence complementary to the mRNA encoded by the nucleic acid sequence shown in SEQ ID NO: 4, SEQ ID NO: 5 or SEQ ID NO: 6, or the RNAi agent comprises, or consists of, a nucleic acid sequence having a sequence identity of about 80% or more with respect to the mRNA encoded by the nucleic acid sequence shown in SEQ ID NO: 4, SEQ ID NO: 5 or SEQ ID NO:

6. The use according to claim 9.

11. The gene editing agent is selected from the group consisting of TALEN-based agents, ZFN-based agents and CRISPR-based agents. The use according to claim 2.

12. The gene editing agent knocks out or knockdowns the expression of TMIGD2. The use according to claim 11.

13. The cancer is a human hematological malignancy. The use according to claim 1.

14. The human blood malignancies are selected from myeloid tumors, acute myeloid leukemia (AML), AML with recurrent genetic abnormalities, AML with myelodysplasia-related changes, AML related to treatment, acute leukemia showing an unclear differentiation lineage, myeloproliferative tumors, essential thrombocythemia, polycythemia vera, myelofibrosis (MF), primary myelofibrosis, systemic mastocytosis, myelodysplastic syndrome (MDS), myeloproliferative / myelodysplastic syndrome, chronic myeloid leukemia, chronic neutrophilic leukemia, chronic eosinophilic leukemia, myelodysplastic syndrome (MDS), sideroblastic refractory anemia, refractory cytopenia with multilineage dysplasia, refractory anemia with excess blasts (type 1), refractory anemia with excess blasts (type 2), MDS with isolated del(5q), unclassifiable MDS, myeloproliferative / myelodysplastic syndrome, chronic myelomonocytic leukemia, atypical chronic myeloid leukemia, juvenile myelomonocytic leukemia, unclassifiable myeloproliferative / myelodysplastic syndrome, lymphoid tumors, precursor lymphoid tumors, B lymphoblastic leukemia, B lymphoblastic lymphoma, T lymphoblastic leukemia, T lymphoblastic lymphoma, mature B cell tumors, diffuse large B cell lymphoma, primary central nervous system lymphoma, primary mediastinal B cell lymphoma, Burkitt lymphoma / leukemia, follicular lymphoma, chronic lymphocytic leukemia, small lymphocytic lymphoma, B cell prolymphocytic leukemia, lymphoplasmacytic lymphoma, Waldenström macroglobulinemia, mantle cell lymphoma, marginal zone lymphoma, post-transplant lymphoproliferative disorder, HIV-related lymphoma, primary effusion lymphoma, intravascular large B cell lymphoma, primary cutaneous B cell lymphoma, hairy cell leukemia, multiple myeloma, monoclonal gammopathy of undetermined significance (MGUS), smoldering multiple myeloma or solitary plasmacytoma (bone and extramedullary), the use according to claim 13.

15. An anti-TMIGD2 antibody or an antigen-binding fragment thereof, (a) a heavy chain variable region comprising GYTFTSYDIN (SEQ ID NO: 24), WIYPGDGSTNYNEKFKG (SEQ ID NO: 25), and ARRLRYYFDY (SEQ ID NO: 26) ; and / or a light chain variable region comprising RASQDIRNYLN (SEQ ID NO: 32), YTSRLHS (SEQ ID NO: 33), and QQVNTLPWT (SEQ ID NO: 34); or (b) GYSITSDYAWN (SEQ ID NO: 56), YITYSGSTSYNPSLKS (SEQ ID NO: 57), and ​ The heavy chain variable region containing ARSGYRDDAMDY (SEQ ID NO: 58); and / or The light chain variable region containing KSSQSLSSNNQKNYLA (SEQ ID NO: 64), FASTRES (SEQ ID NO: 65), and QQHYRTPLT (SEQ ID NO: 66) An anti-TMIGD2 antibody or an antigen-binding fragment thereof comprising Claims 16 The antibody according to claim 15, wherein the antibody comprises (a) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 23; and / or a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 31; or (b) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 55; and / or a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 63 An anti-TMIGD2 antibody or an antigen-binding fragment thereof according to claim 15. ​

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