TREATING CANCER BY BLOCKING THE INTERACTION OF VISTA AND ITS BINDING PARTNER

1 . A method of disrupting an interaction between VISTA and LRIG1, comprising:

contacting a plurality of cells comprising a LRIG1-expressing cell, a VISTA-expressing cell, or a combination thereof with an antibody that specifically binds to LRIG1.

2 . The method of claim 1 , wherein the LRIG1-VISTA interaction is reduced to less than 80%, less than 78%, less than 70%, less than 72%, less than 66%, less than 60%, less than 56%, less than 54%, less than 52%, less than 50%, less than 44%, less than 43%, less than 40%, less than 30%, less than 29%, less than 27%, less than 21%, less than 20%, less than 19%, less than 17%, less than 10%, less than 5%, or less than 1%.

3 . The method of claim 1 , wherein the interaction occurs at one or more residues of LRIG1 selected from region 245-260, wherein the residue positions correspond to positions 245-260 of SEQ ID NO: 2.

4 . The method of claim 1 , wherein the interaction occurs at one or more residues of VISTA selected from region 78-90 or 68-92, wherein the residue positions correspond to positions 78-90 or 68-92 of SEQ ID NO: 4.

5 . The method of claim 1 , wherein the antibody binds to at least one amino acid residue within Peptide 54 or Peptide 61.

6 . The method of claim 1 , wherein the antibody comprises a kD of less than 1 nM, 1.2 nM, 2 nM, 5 nM, 10 nM, 13.5 nM, 15 nM, 20 nM, 25 nM, or 30 nM.

7 . The method of claim 1 , wherein the antibody comprises a humanized antibody.

8 . The method of any one of the claims 1 - 7 , wherein the antibody comprises a full-length antibody or a binding fragment thereof.

9 . The method of any one of the claims 1 - 8 , wherein the antibody comprises a bispecific antibody or a binding fragment thereof.

10 . The method of any one of the claims 1 - 9 , wherein the antibody comprises a monovalent Fab′, a divalent Fab2, a single-chain variable fragment (scFv), a diabody, a minibody, a nanobody, a single-domain antibody (sdAb), or a camelid antibody or binding fragment thereof.

11 . The method of claim 1 , wherein the antibody is a humanized antibody comprising six complementarity-determining regions (CDRs) SEQ ID NOs: 81-86.

12 . The method of claim 11 , wherein the humanized antibody comprises a heavy chain variable region (VH) selected from SEQ ID NOs: 87 and 88.

13 . The method of claim 11 , wherein the humanized antibody comprises a light chain variable region (VL) selected from SEQ ID NOs: 89 and 90.

14 . The method of claim 1 , wherein the antibody is mab2, mab4, mab5, or mab6.

15 . The method of any one of the claims 1 - 10 , wherein the antibody comprises an IgG framework.

16 . The method of any one of the claims 1 - 15 , wherein the antibody comprises an IgG1, IgG2, or IgG4 framework.

17 . A method of inducing immune activation, comprising:

contacting a plurality of cells comprising a LRIG1-expressing cell with an antibody under conditions to effect production of a cytokine, thereby inducing immune activation, wherein the antibody specifically binds to LRIG1.

18 . The method of claim 17 , wherein the plurality of cells further comprises a VISTA expressing cell.

19 . The method of claim 18 , wherein the anti-LRIG1 antibody further inhibits or disrupts an interaction of LRIG1 and VISTA.

20 . The method of claim 19 , wherein the LRIG1-VISTA interaction is reduced to less than 80%, less than 78%, less than 70%, less than 72%, less than 66%, less than 60%, less than 56%, less than 54%, less than 52%, less than 50%, less than 44%, less than 43%, less than 40%, less than 30%, less than 29%, less than 27%, less than 21%, less than 20%, less than 19%, less than 17%, less than 10%, less than 5%, or less than 1%.

21 . The method of claim 19 , wherein the interaction occurs at one or more residues of LRIG1 selected from region 245-260, wherein the residue positions correspond to positions 245-260 of SEQ ID NO: 2.

22 . The method of claim 19 , wherein the interaction occurs at one or more residues of VISTA selected from region 78-90 or 68-92, wherein the residue positions correspond to positions 78-90 or 68-92 of SEQ ID NO: 4.

23 . The method of any one of the claims 17 - 22 , wherein the antibody binds to at least one amino acid residue within Peptide 54 or Peptide 61.

24 . The method of any one of the claims 17 - 23 , wherein the antibody comprises a kD of less than 1 nM, 1.2 nM, 2 nM, 5 nM, 10 nM, 13.5 nM, 15 nM, 20 nM, 25 nM, or 30 nM.

25 . The method of any one of the claims 17 - 24 , wherein the antibody comprises a humanized antibody.

26 . The method of any one of the claims 17 - 25 , wherein the antibody comprises a full-length antibody or a binding fragment thereof.

27 . The method of any one of the claims 17 - 26 , wherein the antibody comprises a bispecific antibody or a binding fragment thereof.

28 . The method of any one of the claims 17 - 27 , wherein the antibody comprises a monovalent Fab′, a divalent Fab2, a single-chain variable fragment (scFv), a diabody, a minibody, a nanobody, a single-domain antibody (sdAb), or a camelid antibody or binding fragment thereof.

29 . The method of any one of the claims 17 - 28 , wherein the antibody is a humanized antibody comprising six complementarity-determining regions (CDRs) SEQ ID NOs: 81-86.

30 . The method of any one of the claims 17 - 29 , wherein the humanized antibody comprises a heavy chain variable region (VH) selected from SEQ ID NOs: 87 and 88.

31 . The method of any one of the claims 17 - 30 , wherein the humanized antibody comprises a light chain variable region (VL) selected from SEQ ID NOs: 89 and 90.

32 . The method of any one of the claims 17 - 31 , wherein the antibody is mab2, mab4, mab5, or mab6.

33 . The method of any one of the claims 17 - 32 , wherein the antibody comprises an IgG framework.

34 . The method of any one of the claims 17 - 33 , wherein the antibody comprises an IgG1, IgG2, or IgG4 framework.

35 . The method of any one of claims 17 - 34 , wherein the cytokine is an interferon.

36 . The method of claim 35 , wherein the interferon is IFNγ.

37 . The method of claim 36 , wherein the antibody results in IFNγ production higher than an isotype antibody.

38 . The method of any one of the claims 17 - 37 , wherein the immune activation comprises a proliferation of CD3+T lymphocytes, CD4+T helper cells, CD8+ cytotoxic T cells, B cells, Natural Killer cells, or a combination thereof.

39 . The method of any one of the claims 17 - 38 , wherein the immune activation comprises an increase in M1 macrophage population within the plurality of cells.

40 . The method of any one of the claims 17 - 39 , wherein the immune activation comprises a decrease in M2 macrophage population within the plurality of cells.

41 . A method of reducing tumor cells within a tumor microenvironment (TME) in a subject, comprising

contacting a plurality of cells located within the TME with an antibody that specifically binds to LRIG1.

42 . The method of claim 41 , wherein the tumor cells are reduced by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, or 90%.

43 . The method of claim 41 , wherein the subject is diagnosed with a cancer.

44 . The method of claim 43 , wherein the cancer is a solid tumor.

45 . The method of claim 44 , wherein the cancer is breast cancer, colorectal cancer, kidney cancer, liver cancer, or lung cancer.

46 . The method of claim 43 , wherein the cancer is a hematologic malignancy.

47 . The method of any one of the claims 43 - 46 , wherein the cancer is a metastatic cancer.

48 . The method of any one of the claims 43 - 46 , wherein the cancer is a relapsed or refractory cancer.

49 . The method of any one of the claims 41 - 48 , wherein the antibody is formulated for systemic administration.

50 . The method of any one of the claims 41 - 48 , wherein the antibody is formulated for parenteral administration.

51 . The method of any one of the claims 41 - 50 , wherein the antibody is administered in combination with an additional therapeutic agent.

52 . The method of claim 51 , wherein the antibody and the additional therapeutic agent are administered simultaneously.

53 . The method of claim 51 , wherein the antibody and the additional therapeutic agent are administered sequentially.

54 . The method of claim 53 , wherein the antibody is administered prior to administering the additional therapeutic agent.

55 . The method of claim 53 , wherein the antibody is administered after administering the additional therapeutic agent.

56 . The method of any one of the claims 51 - 55 , wherein the additional therapeutic agent comprises an immune checkpoint modulator.

57 . The method of any one of the claims 51 - 55 , wherein the additional therapeutic agent comprises a chemotherapeutic agent, targeted therapeutic agent, hormonal therapeutic agent, or a stem cell-based therapeutic agent.

58 . The method of claim 57 , wherein the antibody is administered either prior to or after surgery.

59 . The method of claim 57 , wherein the antibody is administered in conjunction with, before, or after radiation therapy.

60 . The method of any one of the claims 43 - 59 , wherein the anti-LRIG1 antibody further inhibits or disrupts an interaction of LRIG1 and VISTA.

61 . The method of claim 60 , wherein the LRIG1-VISTA interaction is reduced to less than 80%, less than 78%, less than 70%, less than 72%, less than 66%, less than 60%, less than 56%, less than 54%, less than 52%, less than 50%, less than 44%, less than 43%, less than 40%, less than 30%, less than 29%, less than 27%, less than 21%, less than 20%, less than 19%, less than 17%, less than 10%, less than 5%, or less than 1%.

62 . The method of claim 60 , wherein the interaction occurs at one or more residues of LRIG1 selected from region 245-260, wherein the residue positions correspond to positions 245-260 of SEQ ID NO: 2.

63 . The method of claim 60 , wherein the interaction occurs at one or more residues of VISTA selected from region 78-90 or 68-92, wherein the residue positions correspond to positions 78-90 or 68-92 of SEQ ID NO: 4.

64 . The method of any one of the claims 41 - 63 , wherein the antibody binds to at least one amino acid residue within Peptide 54 or Peptide 61.

65 . The method of any one of the claims 41 - 64 , wherein the antibody comprises a kD of less than 1 nM, 1.2 nM, 2 nM, 5 nM, 10 nM, 13.5 nM, 15 nM, 20 nM, 25 nM, or 30 nM.

66 . The method of any one of the claims 41 - 65 , wherein the antibody comprises a humanized antibody.

67 . The method of any one of the claims 41 - 66 , wherein the antibody comprises a full-length antibody or a binding fragment thereof.

68 . The method of any one of the claims 41 - 67 , wherein the antibody comprises a bispecific antibody or a binding fragment thereof.

69 . The method of any one of the claims 41 - 68 , wherein the antibody comprises a monovalent Fab′, a divalent Fab2, a single-chain variable fragment (scFv), a diabody, a minibody, a nanobody, a single-domain antibody (sdAb), or a camelid antibody or binding fragment thereof.

70 . The method of any one of the claims 41 - 69 , wherein the antibody is a humanized antibody comprising six complementarity-determining regions (CDRs) SEQ ID NOs: 81-86.

71 . The method of any one of the claims 41 - 70 , wherein the humanized antibody comprises a heavy chain variable region (VH) selected from SEQ ID NOs: 87 and 88.

72 . The method of any one of the claims 41 - 71 , wherein the humanized antibody comprises a light chain variable region (VL) selected from SEQ ID NOs: 89 and 90.

73 . The method of any one of the claims 41 - 72 , wherein the antibody is mab2, mab4, mab5, or mab6.

74 . The method of any one of the claims 41 - 73 , wherein the antibody comprises an IgG framework.

75 . The method of any one of the claims 41 - 74 , wherein the antibody comprises an IgG1, IgG2, or IgG4 framework.

76 . The method of any one of claims 41 - 75 , further comprising inducing immune activation.

77 . The method of claim 76 , wherein the immune activation comprises production of a cytokine.

78 . The method of claim 77 , wherein the cytokine is an interferon, optionally IFNγ.

79 . The method of any one of the claims 76 - 78 , wherein the immune activation comprises a proliferation of CD3+T lymphocytes, CD4+T helper cells, CD8+ cytotoxic T cells, B cells, Natural Killer cells, or a combination thereof.

80 . The method of any one of the claims 76 - 79 , wherein the immune activation comprises an increase in M1 macrophage population within the plurality of cells.

81 . The method of any one of the claims 76 - 80 , wherein the immune activation comprises a decrease in M2 macrophage population within the plurality of cells.

82 . The method of any one of the claims 41 - 81 , wherein the subject is a human.