Methods of producing enriched populations of tumor-reactive t cells from tumor

Patent metadata

Field	Value
Publication number	US-2016010058-A1
Application number	US-201314771615-A
Country	US
Kind code	A1
Filing date	Apr 30, 2013
Priority date	Mar 1, 2013
Publication date	Jan 14, 2016
Grant date	—

Abstract

Methods of obtaining a cell population enriched for tumor-reactive T cells, the method comprising: (a) obtaining a bulk population of T cells from a tumor sample; (b) specifically selecting CD8 + T cells that express any one or more of TIM-3, LAG-3, 4-1BB, and PD-1 from the bulk population; and (c) separating the cells selected in (b) from unselected cells to obtain a cell population enriched for tumor-reactive T cells are disclosed. Related methods of administering a cell population enriched for tumor-reactive T cells to a mammal, methods of obtaining a pharmaceutical composition comprising a cell population enriched for tumor-reactive T cells, and isolated or purified cell populations are also disclosed.

First claim

1 . A method of obtaining a cell population enriched for tumor-reactive T-cells, the method comprising:
(a) obtaining a bulk population of T cells from a tumor sample; (b) specifically selecting CD8 + T cells that express any one or more of TIM-3. LAG-3, 4-1BB, and PD-1 from the bulk population; and (c) separating the cells selected in (h) from unselected cells to obtain a cell population enriched for tumor-reactive T cells.

2 . A method of obtaining a pharmaceutical composition comprising a cell population enriched for tumor-reactive T cells, the method comprising:
(a) obtaining a bulk population of T cells from a tumor sample; (b) specifically selecting CD8 + T cells that express any one or more of TIM-3, LAG-3, 4-1BB, and PD-1 from the bulk population; (c) separating the cells selected in (b) from unselected cells to obtain a cell population enriched for tumor-reactive T cells; and (d) combining the cell population enriched for tumor-reactive T cells with a pharmaceutically acceptable carrier to obtain a pharmaceutical composition comprising a cell population enriched for tumor-reactive T cells.

3 . The method of claim 1 , wherein (b) comprises specifically selecting CD8 + T cells that express TIM-3 from the bulk population.

4 . The method of claim 1 , wherein (b) comprises specifically selecting CD8 + T cells that express LAG-3 from the bulk population.

5 . The method of claim 1 , wherein (b) comprises specifically selecting CD8 + T cells that express 4-1BB from the bulk population.

6 . The method of claim 1 , wherein (b) comprises specifically selecting CD8 + T cells that express PD-1 from the bulk population.

7 . The method of claim 1 , wherein (b) comprises specifically selecting CD8 + T cells that are (i) 4-1BB + /PD-1 + , (ii) 4-1BB − /PD-1 + , and/or (iii) 4-1BB 30 /PD-1 − from the bulk population.

8 . The method of claim 1 , wherein (b) comprises specifically selecting CD8 + T cells that are (i) LAG-3 + /PD-1 + , (ii) LAG-3 − /PD-1 + , and/or (iii) LAG-3 + /PD-1 − from the bulk population.

9 . The method of claim 1 , wherein (b) comprises specifically selecting CD8 + T cells that are (i) TIM-3 + /PD-1 + , (ii) TIM-3 − /PD-1 + , or (iii) TIM-3 + /PD-1 − from the bulk population.

10 . The method of claim 1 , wherein (b) comprises specifically selecting CD8 + T cells that are (i) TIM-3 + /LAG-3 + , (ii) TIM-3 − /LAG-3 + , or (iii) TIM-3 + /LAG-3 − from the bulk population.

11 . The method of claim 1 , wherein (b) comprises specifically selecting CD8 + T cells that are (i) 4-1BB + /LAG-3 + , (ii) 4-1BB − /LAG-3 + , or (iii) 4-1BB + /LAG-3 − from the bulk population.

12 . The method of claim 1 , wherein (b) comprises specifically selecting CD8 + T cells that are (i) 4-1BB + /TIM-3 + , (ii) 4-1BW/TIM-3 ÷ , or (iii) 4-1BB + /TIM-3 − from the bulk population.

13 . The method of claim 1 , wherein the cell population enriched for tumor-reactive T cells is obtained without screening for autologous tumor recognition.

14 . The method of claim 1 , wherein the bulk population of T cells is not non-specifically stimulated prior to (b).

15 . The method of claim 1 , further comprising expanding the numbers of T cells in the enriched cell population obtained in (c).

16 . The method of claim 1 , further comprising culturing the enriched cell population obtained in (c) in the presence of any one or more of TWS 119, interleukin (IL-21), IL-12, IL-15, IL-7, transforming growth factor (TGF) beta, and AKT inhibitor (AKTi).

17 . The method of claim 1 , further comprising stimulating the enriched cell population obtained in (c) with a cancer antigen and/or with autologous tumor cells.

18 . The method of claim 1 , further comprising transducing or transfecting the cells of the enriched population obtained in (c) with a nucleotide sequence encoding any one or more of IL-12, IL-7, IL-15, IL-2, IL-21, mir155, and anti-PD-1 siRNA.

19 . An isolated or purified cell population enriched for tumor-reactive T cells obtained by the method of claim 1 .

20 . An isolated or purified cell population comprising any one or more of:
(a) CD8 + /4-1BB + /PD-1 + T cells, (b) CD8 + /4-1BB 31 /PD-1 + T cells, (c) CD8 + /4-1BB + /PD-1 − T cells, (d) CD8 + /LAG-3 + /PD-1 + T cells, (e) CD8 + /LAG-3 31 /PD-1 + T cells, (f) CD8 + /LAG-3 + /PD-1 − T cells, (g) CD8 + /TIM-3 + /PD-1 + T cells, (h) CD8 + /TIM-3 − /PD-1 + T cells, (i) CD8 + /TIM-3 + /PD-1 − T cells, (j) CD8 + /TIM-3 + /LAG-3 + T cells, (k) CD8 + /TIM-3 − /LAG-3 + T cells, (l) CD8 + /TIM-3 + /LAG-3 − T cells, (m) CD8 + /4-1BB + /LAG-3 + T cells, (n) CD8 + /4-1BB − /LAG-3 + T cells, (o) CD8 + /4-1BB + , LAG-3 − T cells, (p) CD8 + /4-1BB + /TIM-3 + T cells, (q) CD8 + /4-1BB − /TIM-3 + T cells, and (r) CD8 + /4-1BB + /TIM-3 − T cells, wherein the cell population is enriched for tumor-rcactive T cells.

21 . The isolated or purified cell population of claim 20 comprising:
(a) CD8 + /4-1BB + /PD-1 + T cells,
(b) CD8 + /4-1BB 31 /PD-1 + T cells,
(c) CD8 + /4-1BB + /PD-1 − T cells,
(d) CD8 + /LAG-3 + /PD-1 + T cells,
(e) CD8 + /LAG-3 − /PD-1 30 T cells,
(f) CD8 + /LAG-3 + /PD-1 − T cells,
(g) CD8 + /TIM-3 + /PD-1 + T cells,
(h) CD8 + /TIM-3 − /PD-1 + T cells,
(i) CD8 + /TIM-3 30 /PD-1 − T cells,
(j) CD8 + /TIM-3 + /LAG-3 + T cells,
(k) CD8 + /TIM-3 − /LAG-3 + T cells,
(l) CD8 + /TIM-3 + /LAG-3 − T cells,
(m) CD8 + /4BB + /LAG-3 + T cells,
(n) CD8 + /4-1BB − /LAG-3 + T cells,
(o) CD8 + /4-1BB + /LAG-3 − T cells,
(p) CD8 + /4-1BB + /TIM-3 + T cells,
(q) CD8 + /4-1BB − /TIM-3 + T cells, or
(r) CD8 + /4-BB + /TIM-3 − T cells.

22 . A method of administering a cell population enriched for tumor-reactive T cells to a mammal, the method comprising:
(a) obtaining a bulk population of T cells from a tumor sample; (b) specifically selecting CD8 + T cells that express any one or more of TIM-3, LAG-3, 4-1BB, and PD-1 from the bulk population; (c) separating the cells selected in (b) from unselected cells to obtain a cell population enriched for tumor-reactive T cells; and administering the cell population enriched for tumor-reactive T cells to the mammal.

23 . The method of claim 22 , wherein (b) comprises specifically selecting CD8 + T cells that express TIM-3 from the bulk population.

24 . The method of claim 22 , wherein (b) comprises specifically selecting CD8 + T cells that express LAG-3 from the bulk population.

25 . The method of claim 22 , wherein (b) comprises specifically selecting CD8 + T cells that express 4-1BB from the bulk population.

26 . The method of claim 22 , wherein (b) comprises specifically selecting CD8 + T cells that express PD-1 from the bulk population.

27 . The method of claim 22 , wherein (b) comprises specifically selecting CD8 + T cells that are (i) 4-1BB + /PD-1 + , (ii) 4-1BB − /PD-1 + , and/or (iii) 4-1BB + /PD-1 − from the bulk population.

28 . The method, of claim 22 , wherein (b) comprises specifically selecting CD8 + T cells that are (i) LAG-3 + /PD-1 + , (ii) LAG-3 − /PD-1 + , and/or (iii) LAG-3 + /PD-1 − from the bulk population.

29 . The method of claim 22 , wherein (b) comprises specifically selecting CD8 + T cells that are (i) TIM-3 + /PD-1 + , (ii) TIM-3 − /PD-1 + , or (iii) TIM-3 + /PD-1 − from the bulk po

Assignees

• Us Health

Inventors

• Gros Alena
• Rosenberg Steven A

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• A61P35/02

  specific for leukemia · CPC title

• A61K40/4272

  Melan-A/MART · CPC title

• A61K40/4215

  Receptors for tumor necrosis factors [TNF], e.g. lymphotoxin receptor [LTR], CD30 · CPC title

• A61K40/4202

  Receptors, cell surface antigens or cell surface determinants · CPC title