Compositions and Methods for Targeting Stromal Cells for the Treatment of Cancer

• US 20140099340A1
• Filed: 09/30/2013
• Published: 04/10/2014
• Est. Priority Date: 10/01/2012
• Status: Active Grant

• Current Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Original Assignee: The Wistar Institute, Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Inventors: June, Carl H., Albelda, Steven, Pure, Ellen, Wang, Liang-Chuan, Scholler, John
• Granted Patent: US 9,365,641 B2
• Time in Patent Office: Days
• US Class Current: 424/277.1
• CPC Class Codes: A61K 2239/31 characterized by the route of administration
• CPC Class Codes: A61K 2239/38 characterised by the dose, timing or administration schedule
• CPC Class Codes: A61K 2239/55 Lung
• CPC Class Codes: A61K 35/17 Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes when activated by a specific antigen A61K39/00
• CPC Class Codes: A61K 39/0011 Cancer antigens
• CPC Class Codes: A61K 39/4611 T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
• CPC Class Codes: A61K 39/4631 Chimeric Antigen Receptors [CAR]
• CPC Class Codes: A61K 39/46446 Serine proteases, e.g. kallikrein
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 14/7051 T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 14/70517 CD8
• CPC Class Codes: C07K 14/70521 CD28, CD152
• CPC Class Codes: C07K 14/70578 NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95 NGF-receptor C07K14/71, TNF-receptor C07K14/7151
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 16/30 from tumour cells
• CPC Class Codes: C07K 16/40 against enzymes
• CPC Class Codes: C07K 2317/00 Immunoglobulins specific features
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/51 Complete heavy chain or Fd fragment, i.e. VH + CH1
• CPC Class Codes: C07K 2317/515 Complete light chain, i.e. VL + CL
• CPC Class Codes: C07K 2317/622 : Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/73 : Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2319/02 : containing a signal sequence
• CPC Class Codes: C07K 2319/03 : containing a transmembrane segment
• CPC Class Codes: C07K 2319/30 : Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: C07K 2319/70 : containing domain for protein-protein interaction
• CPC Class Codes: C12N 2710/10343 : viral genome or elements thereof as genetic vector

Abstract

The present invention provides compositions and methods for treating cancer in a human. The invention relates to targeting the stromal cell population in a tumor microenvironment. For example, in one embodiment, the invention provides a composition that is targeted to fibroblast activation protein (FAP). The invention includes a chimeric antigen receptor (CAR) which comprises an anti-FAP domain, a transmembrane domain, and a CD3zeta signaling domain.

Claims

1. An isolated nucleic acid sequence encoding a chimeric antigen receptor (CAR), wherein the CAR comprises an antigen binding domain, a transmembrane domain, a costimulatory signaling region, and a CD3 zeta signaling domain, wherein the antigen binding domain binds to a stromal cell antigen.

2. The isolated nucleic acid sequence of claim 1, wherein the antigen binding domain is an antibody or an antigen-binding fragment thereof.

3. The isolated nucleic acid sequence of claim 2, wherein the antigen-binding fragment is a Fab or a scFv.

4. The isolated nucleic acid sequence of claim 1, wherein the stromal cell antigen is expressed on a stromal cell present in a tumor microenvironment.

5. The isolated nucleic acid sequence of claim 4, wherein the tumor is a carcinoma.

6. The isolated nucleic acid of claim 1, wherein the stromal cell antigen is fibroblast activation protein (FAP).

7. The isolated nucleic acid sequence of claim 1, wherein the costimulatory signaling region comprises the intracellular domain of a costimulatory molecule selected from the group consisting of CD27, CD28, 4-1BB, OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, and any combination thereof.

8. The isolated nucleic acid sequence of claim 1, wherein the nucleic acid sequence comprises SEQ ID NO:

1.

9. An isolated chimeric antigen receptor (CAR) comprising an antigen binding domain, a transmembrane domain, a costimulatory signaling region, and a CD3 zeta signaling domain, wherein the antigen binding domain binds to a stromal cell antigen.

10. The isolated CAR of claim 9, wherein the antigen binding domain is an antibody or an antigen-binding fragment thereof.

11. The isolated CAR of claim 10, wherein the antigen-binding fragment is a Fab or a scFv.

12. The isolated CAR of claim 9, wherein the stromal cell antigen is expressed on a stromal cell present in a tumor microenvironment.

13. The isolated CAR of claim 12, wherein the tumor is a carcinoma.

14. The isolated CAR of claim 9, wherein the stromal cell antigen is FAP.

15. The isolated CAR of claim 9, wherein the costimulatory signaling region comprises the intracellular domain of a costimulatory molecule selected from the group consisting of CD27, CD28, 4-1BB, OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, and any combination thereof.

16. The isolated CAR of claim 9, wherein the CAR is encoded by a nucleic acid sequence comprising SEQ ID NO:

1.

17. A cell comprising a nucleic acid sequence encoding a chimeric antigen receptor (CAR), the CAR comprising an antigen binding domain, a transmembrane domain, a costimulatory signaling region, and a CD3 zeta signaling domain, wherein the antigen binding domain binds to a stromal cell antigen.

18. The cell of claim 17, wherein the antigen binding domain is an antibody or an antigen-binding fragment thereof.

19. The cell of claim 18, wherein the antigen-binding fragment is a Fab or a scFv.

20. The cell of claim 17, wherein the stromal cell antigen is expressed on a stromal cell present in a tumor microenvironment.

21. The cell of claim 20, wherein the tumor is a carcinoma.

22. The cell of claim 17, wherein the stromal cell antigen is FAP.

23. The cell of claim 17, wherein the costimulatory signaling region comprises the intracellular domain of a costimulatory molecule selected from the group consisting of CD27, CD28, 4-1BB, OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, and any combination thereof.

24. The cell of claim 17, wherein the cell is selected from the group consisting of a T cell, a natural killer (NK) cell, a cytotoxic T lymphocyte (CTL), and a regulatory T cell.

25. The cell of claim 17, wherein the nucleic acid sequence comprises SEQ ID NO:

1.

26. A vector comprising a nucleic acid sequence encoding a chimeric antigen receptor (CAR), wherein the CAR comprises an antigen binding domain, a transmembrane domain, a costimulatory signaling region, and a CD3 zeta signaling domain, wherein the antigen binding domain binds to a stromal cell antigen.

27. The vector of claim 26, wherein the antigen binding domain is an antibody or an antigen-binding fragment thereof.

28. The vector of claim 27, wherein the antigen-binding fragment is a Fab or a scFv.

29. The vector of claim 26, wherein the stromal cell antigen is expressed on a stromal cell present in a tumor microenvironment.

30. The vector of claim 29, wherein the tumor is a carcinoma.

31. The vector of claim 26, wherein the stromal cell antigen is FAP.

32. The vector of claim 26, wherein the costimulatory signaling region comprises the intracellular domain of a costimulatory molecule selected from the group consisting of CD27, CD28, 4-1BB, OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, and any combination thereof.

33. The vector of claim 26, wherein the nucleic acid sequence comprises SEQ ID NO:

1.

34. A method for stimulating a T cell-mediated immune response to a stromal cell population in a mammal, the method comprising administering to a mammal an effective amount of a cell genetically modified to express a CAR, wherein the CAR comprises an antigen binding domain, a transmembrane domain, a costimulatory signaling region, and a CD3 zeta signaling domain, and wherein the antigen binding domain is selected to specifically recognize the stromal cell population.

35. The method of claim 34, wherein the antigen binding domain binds to FAP.

36. A method of treating a human with cancer, the method comprising administering to the human a T cell genetically engineered to express a CAR, wherein the CAR comprises an antigen binding domain, a transmembrane domain, a costimulatory signaling region, and a CD3 zeta signaling domain, wherein the antigen binding domain binds to a stromal cell antigen.

37. The method of claim 36, wherein the antigen binding domain is an antibody or an antigen-binding fragment thereof.

38. The method of claim 37, wherein the antigen-binding fragment is a Fab or a scFv.

39. The method of claim 36, wherein the stromal cell antigen is expressed on a stromal cell present in a tumor microenvironment.

40. The method of claim 39, wherein the tumor is a carcinoma.

41. The method of claim 36, wherein the stromal cell antigen is FAP.

42. The method of claim 36, wherein the costimulatory signaling region comprises the intracellular domain of a costimulatory molecule selected from the group consisting of CD27, CD28, 4-1BB, OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, and any combination thereof.

43. The method of claim 36, wherein the T cell is an autologous T cell.

44. The method of claim 36, wherein the cancer is treated.

45. The method of claim 36, wherein the CAR is encoded by a nucleic acid sequence comprising SEQ ID NO:

1.

46. A composition comprising an anti-FAP binding domain.

47. The composition of claim 46, wherein the composition is an antibody, or fragment thereof.

48. The composition of claim 46, wherein the composition is encoded by a nucleic acid sequence comprising SEQ ID NO:

3.

49. A composition comprising a cell comprising a nucleic acid sequence encoding a chimeric antigen receptor (CAR), the CAR comprising an antigen binding domain, a transmembrane domain, a costimulatory signaling region, and a CD3 zeta signaling domain, wherein the antigen binding domain binds to a stromal cell antigen, in combination with an antitumor vaccine.

50. A method of treating cancer in a mammal comprising administering to the mammal a cell comprising a nucleic acid sequence encoding a chimeric antigen receptor (CAR), the CAR comprising an antigen binding domain, a transmembrane domain, a costimulatory signaling region, and a CD3 zeta signaling domain, wherein the antigen binding domain binds to a stromal cell antigen, and an antitumor vaccine.

51. The method of claim 50, wherein the cell and the antitumor vaccine are co-administered to the mammal.

52. The method of claim 50, wherein the cell and the antitumor vaccine are administered to the mammal separately.