GPR15-mediated homing and uses thereof

• US 9,376,663 B2
• Filed: 04/02/2014
• Issued: 06/28/2016
• Est. Priority Date: 04/02/2013
• Status: Active Grant

• Current Assignee: New York University
• Original Assignee: New York University
• Inventors: Littman, Dan R., Kim, Sang-Won
• Primary Examiner(s): Gamett, Daniel C
• Application Number:
  US14/243,290
• Publication Number:
  US 20140294793A1
• Time in Patent Office: 818 Days
• US Class Current: 1/1
• CPC Class Codes: A01K 67/0271 Chimeric vertebrates, e.g. comprising exogenous cells
• 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 35/00 Medicinal preparations containing materials or reaction products thereof with undetermined constitution
• 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 39/4621 immunosuppressive or immunotolerising
• CPC Class Codes: A61K 39/46433 Antigens related to auto-immune diseases; Preparations to induce self-tolerance
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C12N 5/0636 T lymphocytes
• CPC Class Codes: C12N 5/0637 Immunosuppressive T lymphocytes, e.g. regulatory T cells or Treg

Abstract

The present invention is directed to in vitro methods for promoting expression of G-protein coupled receptor 15 (GPR15) on T cells, GPR15+ enriched populations of T cells generated using these methods and compositions thereof, as well as methods of using these T cell populations for therapeutic purposes.

Claims

1. A method for generating a population of CD4+CD3+CD25+T cells that migrates preferentially to the large intestine, the method comprising the steps of:

isolating a population of CD4+CD3+CD25+ T cells from a mammal; and

incubating the population of CD4+CD3+CD25+ T cells in culture medium comprising TGF-β and

IL-21 to produce a population of CD4+CD3+CD25+T cells having enhanced GPR15 expression, wherein the enhanced GPR15 expression confers on the population of CD4+CD3+CD25+T cells an ability to migrate preferentially to the large intestine.

2. The method of claim 1, wherein the culture medium comprises TGF-β

at a concentration of at least 0.1 ng/ml and IL-21 at a concentration of at least 0.1 ng/ml.

3. The method of claim 1, wherein the culture medium comprises TGF-β

at a concentration of about 5-20 ng/ml and IL-21 at a concentration of about 10-20 ng/ml.

4. The method of claim 1, wherein the culture medium further comprises IL-2.

5. The method of claim 1, further comprising administering the population of CD4+CD3+CD25+T cells having enhanced GPR15 expression and ability to migrate preferentially to the large intestine to the mammal, wherein the mammal is afflicted with an inflammatory condition of the large intestine.

6. The method of claim 5, wherein the inflammatory condition of the large intestine is inflammatory bowel disease, idiopathic colitis, or infectious colitis.

7. The method of claim 6, wherein the inflammatory bowel disease is Crohn'"'"'s disease or ulcerative colitis.

8. The method of claim 1, further comprising anti-CD3/CD28 activating agents in the incubating step.

9. The method of claim 1, wherein the mammal is a human.

10. The method of claim 1, wherein the culture medium does not comprise retinoic acid.

11. A population of CD4+CD3+CD25+T cells having enhanced GPR15 expression and ability to migrate preferentially to the large intestine and a composition thereof further comprising a pharmaceutically acceptable carrier, wherein the population is generated in accordance with the method of claim 1.

13. A method of treating a mammal in need thereof comprising administering the population of claim 11 to the mammal in need thereof, wherein the population of CD4+CD3+CD25+T cells having enhanced GPR15 expression and ability to migrate preferentially to the large intestine is administered to the mammal in need thereof, wherein the mammal in need thereof is afflicted with an inflammatory condition of the large intestine, thereby treating the mammal in need thereof.

12. A population of CD4+CD3+CD25+T cells having enhanced GPR15 expression and ability to migrate preferentially to the large intestine, wherein the population comprises a nucleic acid sequence encoding exogenous GPR15 or a composition of said cells further comprising a pharmaceutically acceptable carrier.

14. A method for generating a population of T cells that migrates preferentially to the large intestine, the method comprising the steps of:

isolating a population of T cells from a mammal and transfecting/transducing/transforming the population of T cells with a nucleic acid sequence encoding GPR15 to produce a population of T cells having enhanced GPR15 expression, wherein the enhanced GPR15 expression confers on the population of T cells an ability to migrate preferentially to the large intestine.

15. The method of claim 14, wherein the mammal is a human.

16. The method of claim 14, further comprising transfecting/transducing the population of T cells with nucleic acid sequences encoding integrin α

4 and integrin β

7 to confer enhanced expression of integrin α

4β

7 heterodimer to the T cell population.

17. The method of claim 14 or 16, wherein the population of T cells is isolated from whole blood or buffy coats of adults.

18. The method of claim 14 or 16, wherein the population of T cells is activated prior to the transfecting/transducing/transforming step.

19. The method of claim 18, wherein the population of T cells is activated using anti-CD3/CD28 activating agents.

20. The method of claim 14 or 16, wherein the population of T cells comprises CD4+CD3+CD25+T cells, CD4+CD3+CD25+CD127−

T cells, CD4+CD3+CD25−

T cells, or CD8+CD3+ T cells and the transfecting/transducing produces a population of CD4+CD3+CD25+T cells, CD4+CD3+CD25+CD127−

T cells, CD4+CD3+ effector T cells, or CD8+CD3+ effector T cells, respectively, having enhanced GPR15 expression and ability to migrate preferentially to the large intestine.

21. The method of claim 20, further comprising administering to a mammal the population of CD4+CD3+CD25+T cells or CD4+CD3+CD25+CD127−

T cells having enhanced GPR15 expression and ability to migrate preferentially to the large intestine, wherein the mammal is afflicted with an inflammatory condition of the large intestine.

22. The method of claim 21, wherein the inflammatory condition of the large intestine is inflammatory bowel disease, idiopathic colitis, or infectious colitis.

23. The method of claim 22, wherein the inflammatory bowel disease is Crohn'"'"'s disease or ulcerative colitis.

24. The method of claim 20, further comprising administering to a mammal the population of CD4+CD3+ effector T cells or CD8+CD3+ effector T cells having enhanced GPR15 expression and ability to migrate preferentially to the large intestine, wherein the mammal is afflicted with a cancer or an infection of the large intestine.

25. The method of claim 24, wherein the cancer is colon cancer or a cancer that has metastasized to the large intestine.

26. The method of claim 24, wherein the infection is a bacterial or viral infection.

27. A method for generating a population of T cells that migrates preferentially to the large intestine, the method comprising the steps of:

isolating a population of T cells from a mammal and introducing a nucleic acid sequence encoding GPR15 into the population of T cells to produce a population of T cells having enhanced GPR15 expression, wherein the enhanced GPR15 expression confers on the population of T cells an ability to migrate preferentially to the large intestine.