METHODS AND COMPOSITIONS RELATING TO EX VIVO CULTURE AND MODULATION OF T CELLS

• US 20190330343A1
• Filed: 01/10/2018
• Published: 10/31/2019
• Est. Priority Date: 01/10/2017
• Status: Active Grant

• Current Assignee: The General Hospital Corporation (Mass General Brigham, Inc.)
• Original Assignee: The General Hospital Corporation (Mass General Brigham, Inc.)
• Inventors: MAUS, Marcela V., BEDOYA, Felipe
• Granted Patent: US 11,613,574 B2
• Time in Patent Office: Days
• CPC Class Codes: A61K 2039/5154 Antigen presenting cells [APCs], e.g. dendritic cells or macrophages
• 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 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: C07K 14/7051 T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 14/70517 CD8
• CPC Class Codes: C07K 16/2803 against the immunoglobulin superfamily
• CPC Class Codes: C07K 16/2809 against the T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 16/2818 against CD28 or CD152
• CPC Class Codes: C07K 16/2875 against the NGF/TNF superfamily, e.g. CD70, CD95L, CD153, CD154 against NGF C07K16/22, against TNF C07K16/241
• CPC Class Codes: C07K 16/2878 against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• 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 2319/33 fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 2740/16043 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 5/0636 T lymphocytes

Abstract

Described herein are methods for producing and utilizing an artificial antigen presenting cell (aAPC). An aAPC is engineered to express a first and second chimeric stimulatory receptor that specifically bind antigen presenting on a T cell of interest. The aAPC as described herein is designed for use in activating and/or expanding a T cell or chimeric antigen receptor (CAR) T cell. Further, this invention relates to methods of treating cancer by administering to a subject in need thereof an aAPC-activated CART cell.

Claims

1. An artificial antigen presenting cell (aAPC) comprising:

a first chimeric stimulatory receptor (CSR) that binds specifically with a first co-stimulatory polypeptide; and

a second CSR that binds specifically a second co-stimulatory polypeptide.

3. The aAPC of claim 1 or 2, wherein the aAPC has been engineered to lack an expressible low-density lipoprotein receptor (LDLR) gene.

4. The aAPC of claim 1 or 2, wherein the aAPC comprises a deletion in the native LDLR-encoding nucleic acid sequence.

5. The aAPC of claim 1 or 2, wherein the aAPC comprises a deletion of the native LDLR-encoding nucleic acid sequence.

6. The aAPC of claim 1 or 2, wherein the aAPC is viable but non-dividing.

7. The aAPC of claim 1 or 2, wherein the aAPC has been gamma-irradiated.

8. The aAPC of claim 1 or 2, further comprising a T-cell target molecule.

9. The aAPC of claim 8, wherein the T-cell target molecule is CD19, BCMA, CD37, SLAMF7, EGFR, or EGFR variant III.

10. The aAPC of claim 9, wherein the T-cell target molecule is CD19.

11. The aAPC of claim 8, wherein the T-cell target molecule is any molecule that can be targeted by a CAR T cell.

12. The aAPC of claim 1 or 2, wherein the first and second CSRs are expressed on the cell surface of the aAPC.

13. The aAPC of claim 1 or 2, wherein the first and second CSRs are constitutively expressed.

14. The aAPC of claim 1 or 2, wherein the first and second CSRs are encoded by a first recombinant nucleic acid sequence and a second recombinant nucleic acid sequence, respectively.

15. The aAPC of claim 14, wherein the sequence encoding the first and/or second CSR is operatively linked to a constitutive promoter.

16. The aAPC of claim 15, wherein the constitutive promoter is EF1-α

.

17. The aAPC of claim 1 or 2, wherein the aAPC is a human cell.

18. The aAPC of claim 1 or 2, wherein the aAPC is engineered from an erythromyeloid cell.

19. The aAPC of claim 18, wherein the erthromyeloid cell is a K562 cell.

20. The aAPC of claim 1 or 2, wherein the first and second CSRs bind specifically with human CD3 and CD28, respectively.

21. The aAPC of claim 1, wherein the first CSR binds specifically with 4-1BBL, and the second CSR binds specifically with OX40L.

22. A method of expanding or activating a T cell, the method comprising contacting the aAPC of claim 1 with a T cell.

23. A method of treating a cancer, a plasma cell disease or disorder, or an autoimmune disease or disorder in a subject in need thereof, the method comprising:

contacting the aAPC of claim 1 with a CAR T cell, thereby activating the CAR T cell; and

administering the activated CAR T cell to the subject.

24. The method of claim 22 or 23, wherein the contacting step occurs in vitro.

25. The method of claim 22 or 23, wherein the contacting step occurs in suspension.

26. A composition comprising the aAPC of claim 1 or 2 and a T cell.

27. The composition of claim 26, wherein the T cell is a CAR T cell.

28. A composition comprising the activated CAR T cells of the method of claim 22 or 23, formulated for the treatment of cancer.

29. The composition of claim 28, further comprising a pharmaceutically acceptable carrier.

30. The method of claim 23, wherein the cancer is a leukemia, a lymphoma, multiple myeloma, or a solid tumor.

31. The method of claim 30, wherein the leukemia is acute lymphocytic leukemia (ALL) or chronic lymphocytic leukemia (CLL).

32. The method of claim 30, wherein the lymphoma is follicular lymphoma or diffuse large B cell lymphoma (DLBCL).

2. An aAPC comprising:

a first CSR that binds specifically with CD3; and

a second CSR that binds specifically with CD28.