Intracellular genomic transplant and methods of therapy
Genetically modified compositions, such as non-viral vectors and T cells, for treating cancer are disclosed. Also disclosed are the methods of making and using the genetically modified compositions in treating cancer.
1. A method of treating a neoantigen expressing cancer in a human subject, the method comprising:
administering to said human subject an immunosuppressive agent; and
administering to said human subject a population of ex vivo engineered neoantigen targeting human primary cells, wherein said population of ex vivo engineered neoantigen targeting human primary cells comprises CD4+ T cells that comprise:
a) an endonuclease mediated genomic disruption within exon 2 or exon 3 of a cytokine inducible SH2-containing protein gene (CIS-1) sequence, wherein said endonuclease mediated genomic disruption comprises an indel, and wherein said genomic disruption suppresses expression of a protein encoded by the cytokine inducible SH2-containing protein gene (CIS-1); and
b) an exogenous functional neoantigen targeting T cell receptor polypeptide or functional neoantigen targeting fragment thereof, or an exogenous functional neoantigen targeting chimeric antigen receptor polypeptide or functional neoantigen targeting fragment thereof, wherein said neoantigen is expressed by said neoantigen expressing cancer, and
wherein said cancer is selected from the group consisting of: bladder cancer, bone cancer, brain cancer, breast cancer, esophageal cancer, gastrointestinal cancer, liver cancer, lung cancer, ovarian cancer, prostate cancer, anal cancer, rectal cancer, ocular cancer, cancer of the neck, gallbladder cancer, pleural cancer, oral cancer, cancer of the vulva, colon cancer, cervical cancer, fibrosarcoma, kidney cancer, mesothelioma, mastocytoma, nasopharynx cancer, pancreatic cancer, peritoneal cancer, small intestine cancer, stomach cancer, testicular cancer, and thyroid cancer.
2. The method of claim 1 , wherein said population of ex vivo engineered neoantigen targeting human primary cells further comprises cells that comprise a genomic disruption in a T Cell Receptor Alpha Constant or a T Cell Receptor Beta Locus gene sequence.
3. The method of claim 2 , wherein said exogenous functional neoantigen targeting T cell receptor polypeptide or said functional neoantigen targeting fragment thereof, or said exogenous functional neoantigen targeting chimeric antigen receptor polypeptide or said functional neoantigen targeting fragment thereof, is expressed from a DNA molecule integrated into said genomic disruption in said T Cell Receptor Alpha Constant or a T Cell Receptor Beta Locus gene sequence.
4. The method of claim 1 , wherein said population of ex vivo engineered neoantigen targeting human primary cells further comprises: a population of CD8+ T cells, a population of dendritic cells, a population of B cells, a population of natural killer cells, or any combination thereof.
5. The method of claim 1 , wherein said population of ex vivo engineered neoantigen targeting human primary cells further comprises a population of CD8+ T cells.
6. The method of claim 1 , wherein said population of ex vivo engineered neoantigen targeting human primary cells comprises a population of tumor infiltrating lymphocytes.
7. The method of claim 1 , wherein said population of ex vivo engineered neoantigen targeting human primary cells comprises a population of peripheral blood lymphocytes.
8. The method of claim 1 , wherein said population of ex vivo engineered neoantigen targeting human primary cells is autologous to said human subject.
9. The method of claim 1 , wherein said population of ex vivo engineered neoantigen targeting human primary cells is allogeneic to said human subject.
10. The method of claim 1 , wherein said endonuclease mediated genomic disruption is performed by a Clustered Regularly Interspaced Short Palindromic Repeats system.
11. The method of claim 10 , wherein said Clustered Regularly Interspaced Short Palindromic Repeats system comprises a Cas9 protein.
12. The method of claim 1 , wherein said exogenous functional neoantigen targeting T cell receptor polypeptide or said functional neoantigen targeting fragment thereof, or said exogenous functional neoantigen targeting chimeric antigen receptor polypeptide or said functional neoantigen targeting fragment thereof, is expressed from a DNA molecule integrated into said endonuclease mediated genomic disruption in said cytokine inducible SH2-containing protein gene (CIS-1) sequence.
13. The method of claim 1 , wherein said exogenous cell surface receptor or functional fragment thereof comprises an exogenous functional chimeric antigen receptor polypeptide or functional fragment thereof.
14. The method of claim 1 , wherein said exogenous functional neoantigen targeting T cell receptor polypeptide or functional neoantigen targeting fragment thereof, or said exogenous functional neoantigen targeting chimeric antigen receptor polypeptide or functional neoantigen targeting fragment thereof, is introduced into said CD4+ T cells using a viral vector.
15. The method of claim 14 , wherein said viral vector is an adeno-associated virus vector.
16. The method of claim 1 , wherein said genomic disruption results in elimination of said protein encoded by said cytokine inducible SH2-containing protein gene (CIS-1).
17. A method of treating a cancer in a human subject, the method comprising:
administering to said human subject an immunosuppressive agent; and
administering to said subject a population of ex vivo engineered cancer targeting human primary cells, wherein said population of ex vivo cancer targeting engineered human primary cells comprises CD4+ T cells that comprise:
a) an endonuclease mediated genomic disruption within exon 2 or exon 3 of a cytokine inducible SH2-containing protein gene (CIS-1) sequence, wherein said endonuclease mediated genomic disruption comprises an indel, and wherein said genomic disruption suppresses expression of a protein encoded by the cytokine inducible SH2-containing protein gene (CIS-1); and
b) an exogenous cell surface receptor or functional fragment thereof capable of binding a cell of said cancer, and
wherein said cancer is selected from the group consisting of bladder cancer, bone cancer, brain cancer, esophageal cancer, gastrointestinal cancer, liver cancer, lung cancer, ovarian cancer, prostate cancer, anal cancer, rectal cancer, ocular cancer, cancer of the neck, gallbladder cancer, pleural cancer, oral cancer, cancer of the vulva, colon cancer, cervical cancer, fibrosarcoma, kidney cancer, mesothelioma, mastocytoma, nasopharynx cancer, pancreatic cancer, peritoneal cancer, small intestine cancer, stomach cancer, testicular cancer, and thyroid cancer.
18. The method of claim 17 , wherein said exogenous cell surface receptor or functional fragment thereof comprises an exogenous functional T cell receptor or functional fragment thereof.
19. The method of claim 17 , wherein said population of ex vivo engineered cancer targeting human primary cells further comprises: a population of CD8+ T cells, a population of dendritic cells, a population of B cells, a population of natural killer cells, or any combination thereof.
20. The method of claim 17 , wherein said exogenous cell surface receptor or functional fragment thereof is expressed from DNA integrated into said endonuclease mediated genomic disruption.
21. The method of claim 17 , wherein said endonuclease mediated genomic disruption is performed by a Clustered Regularly Interspaced Short Palindromic Repeats system.
22. The method of claim 21 , wherein said Clustered Regularly Interspaced Short Palindromic Repeats system comprises a Cas9 protein.
23. The method of claim 17 , wherein said population of ex vivo engineered cancer targeting human primary cells further comprises cells that comprise a genomic disruption in a T Cell Receptor Alpha Constant or a T Cell Receptor Beta Locus gene sequence.
24. The method of claim 23 , wherein said exogenous cell surface receptor or functional fragment thereof is expressed from DNA integrated into said: cytokine inducible SH2-containing protein gene (CIS-1) sequence, said T Cell Receptor Alpha Constant gene sequence, or said T Cell Receptor Beta Locus gene sequence.
25. The method of claim 17 , wherein said population of ex vivo engineered cancer targeting human primary cells comprises a population of natural killer cells.
26. The method of claim 17 , wherein said population of ex vivo engineered cancer targeting human primary cells comprises a population of tumor infiltrating lymphocytes.
27. The method of claim 17 , wherein said population of ex vivo engineered cancer targeting human primary cells comprises a population of peripheral blood lymphocytes.
28. The method of claim 17 , wherein said population of ex vivo engineered cancer targeting human primary cells is autologous to said human subject.
29. The method of claim 17 , wherein said population of ex vivo engineered cancer targeting human primary cells is allogeneic to said human subject.
30. The method of claim 17 , wherein said exogenous cell surface receptor or functional fragment thereof is introduced into said CD4+ T cells using a viral vector.
31. The method of claim 30 , wherein said viral vector is an adeno-associated virus vector.
32. The method of claim 17 , wherein said genomic disruption results in elimination of said protein encoded by said cytokine inducible SH2-containing protein gene (CIS-1).
33. A method of treating a neoantigen expressing cancer in a human subject, the method comprising:
administering to said human subject an immunosuppressive agent; and
administering to said human subject a population of ex vivo engineered neoantigen targeting human primary cells, wherein said population of ex vivo engineered neoantigen targeting human primary cells comprises CD4+ T cells that comprise:
a) an endonuclease mediated genomic disruption within exon 2 or exon 3 of a cytokine inducible SH2-containing protein gene (CIS-1) sequence, wherein said endonuclease mediated genomic disruption comprises an indel, and wherein said genomic disruption suppresses expression of a protein encoded by the cytokine inducible SH2-containing protein gene (CIS-1); and
b) an exogenous functional neoantigen targeting T cell receptor polypeptide or functional neoantigen targeting fragment thereof, or an exogenous functional neoantigen targeting chimeric antigen receptor polypeptide or functional neoantigen targeting fragment thereof, wherein said neoantigen is expressed by said neoantigen expressing cancer,
and wherein said cancer is selected from the group consisting of: gastrointestinal cancer, breast cancer, and prostate cancer.
34. The method of claim 33 , wherein said population of ex vivo engineered neoantigen targeting human primary cells comprise a genomic disruption in a T Cell Receptor Alpha Constant or a T Cell Receptor Beta Locus gene sequence.
35. The method of claim 34 , wherein said exogenous functional neoantigen targeting T cell receptor polypeptide or said functional neoantigen targeting fragment thereof, or said exogenous functional neoantigen targeting chimeric antigen receptor polypeptide or said functional neoantigen targeting fragment thereof, is expressed from DNA integrated into said genomic disruption in said T Cell Receptor Alpha Constant gene sequence or said T Cell Receptor Beta Locus gene sequence.
36. The method of claim 33 , wherein said population of ex vivo engineered neoantigen targeting human primary cells further comprises: a population of CD8+ T cells, a population of dendritic cells, a population of B cells, a population of natural killer cells, or any combination thereof.
37. The method of claim 33 , wherein said population of ex vivo engineered neoantigen targeting human primary cells comprises a population of tumor infiltrating lymphocytes.
38. The method of claim 33 , wherein said population of ex vivo engineered neoantigen targeting human primary cells comprises a population of peripheral blood lymphocytes.
39. The method of claim 33 , wherein said population of ex vivo engineered neoantigen targeting human primary cells is autologous to said human subject.
40. The method of claim 33 , wherein said population of ex vivo engineered neoantigen targeting human primary cells is allogeneic to said human subject.
41. The method of claim 33 , wherein said exogenous functional neoantigen targeting T cell receptor polypeptide or said functional neoantigen targeting fragment thereof, or said exogenous functional neoantigen targeting chimeric antigen receptor polypeptide or said functional neoantigen targeting fragment thereof, is expressed from a DNA molecule integrated into said endonuclease mediated genomic disruption in said cytokine inducible SH2-containing protein gene (CIS-1) sequence.
42. The method of claim 33 , wherein said endonuclease mediated genomic disruption is performed by a Clustered Regularly Interspaced Short Palindromic Repeats system.
43. The method of claim 42 , wherein said Clustered Regularly Interspaced Short Palindromic Repeats system comprises a Cas9 protein.
44. The method of claim 33 , wherein said exogenous functional neoantigen targeting T cell receptor polypeptide or functional neoantigen targeting fragment thereof, or said exogenous functional neoantigen targeting chimeric antigen receptor polypeptide or functional neoantigen targeting fragment thereof, is introduced into said CD4+ T cells using a viral vector.
45. The method of claim 44 , wherein said viral vector is an adeno-associated virus vector.
46. The method of claim 33 , wherein said genomic disruption results in elimination of said protein encoded by said cytokine inducible SH2-containing protein gene (CIS-1).