BINDING PROTEINS SPECIFIC FOR RAS NEOANTIGENS AND USES THEREOF
The present disclosure provides compositions and methods for targeting a Ras antigen to, for example, treat or prevent cancer. Disclosed embodiments include binding proteins, such as a T cell receptor or a chimeric antigen receptor, that bind to a Ras antigen:HLA complex. Polynucleotides encoding such binding protein can introduced into a host cell, such as a T cell, and the cell can be used in immunotherapy for treating various cancers. Also provided are immunogenic polypeptides that can be useful to, for example, induce an immune response against a mutated Ras or to identify a binding protein that binds to a Ras antigen.
1 - 30 . (canceled)
31 . A method of treating a disease or disorder or preventing a relapse of a disease or disorder associated with a G12-mutant KRAS, NRAS, or HRAS mutation in a subject in need thereof, comprising
administering to said subject an effective amount of a cell population comprising a T cell comprising a membrane protein with a human or humanized extracellular binding domain comprising a T-cell receptor Vα and Vβ region configured to bind to a peptide:HLA complex comprising a G12-mutant KRAS, NRAS, or HRAS peptide, wherein said peptide:HLA complex is HLA-A*11 serotype restricted; wherein said subject is HLA-A*11:01 positive.
32 . The method of claim 31 , wherein said disease or disorder is positive for G12-mutant KRAS, NRAS, or HRAS.
33 . The method of claim 32 , wherein said disease or disorder is positive for G12 mutant KRAS.
34 . The method of claim 33 , wherein said disease or disorder is positive for G12V mutant KRAS.
35 . The method of claim 31 , wherein said cell population further comprises both CD4+ and CD8+ T cells comprising said membrane protein.
36 . The method of claim 31 , wherein said cell population further comprises T cells comprising said membrane protein and (i) a transgenic polynucleotide encoding a polypeptide that comprises an extracellular portion of a CD8 co-receptor α chain; (ii) a transgenic polynucleotide encoding a polypeptide that comprises an extracellular portion of a CD8 co-receptor β chain; or (iii) a transgenic polynucleotide encoding (1) a polypeptide that comprises an extracellular portion of a CD8 co-receptor α chain and (2) a polypeptide that comprises an extracellular portion of a CD8 co-receptor β chain.
37 . The method of claim 31 , wherein said cell population further comprises T cells comprising said membrane protein and a chromosomal gene knockout of a TCR constant region.
38 . The method of claim 31 , wherein a presence of said G12-mutant KRAS, NRAS, or HRAS mutation has been determined in said subject prior to said administering.
39 . The method of claim 31 , wherein said T cell is configured to undergo an antigen-specific T-cell response to said peptide:HLA complex.
40 . The method of claim 31 , wherein said T cell is configured to produce IFN-γ when in the presence of said peptide:HLA complex.
41 . The method of claim 31 , wherein said T cell is configured to have elevated CD137 expression when in the presence of said peptide:HLA complex as compared to CD137 expression in a T cell not expressing said membrane protein.
42 . The method of claim 31 , wherein the T cell does not substantially produce IFN-γ when in the absence of said peptide:HLA complex.
43 . The method of claim 31 , wherein said extracellular binding domain comprises a framework region derived from a human TRBV, TRBD, TRBJ, TRAV, or TRAJ gene segment.
44 . The method of claim 31 , wherein said membrane protein does not comprise a cytoplasmic signaling domain fused thereto.
45 . The method of claim 31 , wherein said membrane protein has a log 10 EC50 for the peptide of less than −8.0, optionally about −8.5 or less, further optionally about −8.5, about −8.6, about −8.7, about −8.8, about −8.9, about −9, about −9.1, or about −9.2
46 . The method of claim 31 , wherein said T cell is a CD8+ T cell, a CD4− CD8− double negative T cell, a γδ T cell, or a natural killer T cell.
47 . The method of claim 31 , wherein said T cell comprises a heterologous polynucleotide sequence encoding a TCR α chain and a TCR β chain separated by a sequence encoding a self-cleaving peptide.
48 . The method of claim 47 , wherein said self-cleaving peptide is a P2A, T2A, F2A, or E2A peptide, or any combination thereof.
49 . The method of claim 31 , further comprising determining the HLA genotype of said subject prior to said administering.
50 . The method of claim 31 , wherein said cell population comprises CD8+ or CD4+ T cells comprising said membrane protein.
51 . The method of claim 31 , wherein said disease or disorder comprises a cancer.
52 . The method of claim 51 , wherein said cancer comprises a solid cancer.
53 . The method of claim 51 , wherein said cancer comprises a hematological malignancy.
54 . the method of claim 52 , wherein said solid cancer comprises a pancreatic, lung, or colorectal cancer.
55 . The method of claim 31 , wherein said T cell is autologous to said subject.
56 . The method of claim 31 , wherein said T cell is allogenic to said subject.
57 . The method of claim 31 , wherein said effective amount of said cell population is from about 10 4 cells/kg to about 10 11 cells/kg.
58 . The method of claim 31 , wherein said subject has received lymphodepleting chemotherapy prior to said administration.
59 . A method of treating a disease or disorder or preventing a relapse of a disease or disorder associated with a G12-mutant KRAS, NRAS, or HRAS mutation in a subject in need thereof, comprising:
administering to said subject an effective amount of a cell population comprising a T cell comprising a membrane protein with a human or humanized extracellular binding domain that is configured to bind to a peptide:HLA complex comprising a G12-mutant KRAS, NRAS, or HRAS peptide, wherein said peptide:HLA complex is HLA-A*11 serotype restricted, wherein said cell population further comprises T cells comprising said membrane protein and (i) a transgenic polynucleotide encoding a polypeptide that comprises an extracellular portion of a CD8 co-receptor α chain; (ii) a transgenic polynucleotide encoding a polypeptide that comprises an extracellular portion of a CD8 co-receptor β chain; or (iii) a transgenic polynucleotide encoding (1) a polypeptide that comprises an extracellular portion of a CD8 co-receptor α chain and (2) a polypeptide that comprises an extracellular portion of a CD8 co-receptor β chain.
60 . The method of claim 59 , wherein said disease or disorder is positive for G12 mutant KRAS.
61 . The method of claim 60 , wherein said disease or disorder is positive for G12V mutant KRAS.
62 . The method of claim 59 , wherein said cell population further comprises both CD4+ and CD8+ T cells comprising said membrane protein.
63 . The method of claim 59 , wherein said cell population further comprises T cells comprising said membrane protein and a chromosomal gene knockout of a TCR constant region.
64 . The method of claim 59 , wherein a presence of said G12-mutant KRAS, NRAS, or HRAS mutation has been determined in said subject prior to said administering.
65 . The method of claim 59 , wherein said extracellular binding domain comprises a T-cell receptor Vα and Vβ region.
66 . The method of claim 59 , wherein said T cell comprises a heterologous polynucleotide sequence encoding a T-cell receptor α chain and a T-cell receptor β chain separated by a sequence encoding a self-cleaving peptide.
67 . The method of claim 66 , wherein said self-cleaving peptide is a P2A, T2A, F2A, or E2A peptide, or any combination thereof.
68 . The method of claim 59 , wherein said subject is HLA-A*11:01 positive.
69 . A method of treating a disease or disorder or preventing a relapse of a disease or disorder associated with a G12-mutant KRAS, NRAS, or HRAS mutation in a subject in need thereof, comprising:
administering to said subject an effective amount of a cell population comprising a T cell comprising a membrane protein with a human or humanized extracellular binding domain that is configured to bind to a peptide:HLA complex comprising a G12-mutant KRAS, NRAS, or HRAS peptide, wherein said peptide:HLA complex is HLA-A*11 serotype restricted, wherein said cell population further comprises both CD4+ and CD8+ T cells comprising said membrane protein.
70 . The method of claim 69 , wherein said disease or disorder is positive for G12 mutant KRAS.
71 . The method of claim 70 , wherein said disease or disorder is positive for G12V mutant KRAS.
72 . The method of claim 69 , wherein a presence of said G12-mutant KRAS, NRAS, or HRAS mutation has been determined in said subject prior to said administering.
73 . The method of claim 69 , wherein said extracellular binding domain comprises a T-cell receptor Vα and Vβ region.
74 . The method of claim 69 , wherein said T cell comprises a heterologous polynucleotide sequence encoding a T-cell receptor α chain and a T-cell receptor β chain separated by a sequence encoding a self-cleaving peptide.
75 . The method of claim 74 , wherein said self-cleaving peptide is a P2A, T2A, F2A, or E2A peptide, or any combination thereof.
76 . The method of claim 69 , wherein said subject is HLA-A*11:01 positive.
77 . The method of claim 69 , further comprising administering a cell population comprising about a 1:1 ratio of CD4+ to CD8+ cells comprising said membrane protein.