Natural killer cells with enhanced activity

Patent metadata

Field	Value
Publication number	US-2024010980-A1
Application number	US-202118249991-A
Country	US
Kind code	A1
Filing date	Oct 27, 2021
Priority date	Oct 27, 2020
Publication date	Jan 11, 2024
Grant date	—

Abstract

Engineered immune cells, optionally natural killer (NK) cells, comprising an inactivating mutation for an endogenous A-Disintegrin-And-Metalloproteinase 17 (ADAM 17) gene. ADAM17-deficient immune cells produced from stem cells, optionally, induced pluripotent stem cells (iPSCs), having enhanced antibody-dependent cellular cytotoxicity (ADCC). Pharmaceutical compositions comprising said engineered immune cells. Methods for making the engineered immune cell and pharmaceutical compositions, and methods of use are provided.

First claim

1 . An engineered immune cell, comprising an inactivating mutation in an endogenous A-Disintegrin-And-Metalloproteinase 17 (ADAM17) gene.

2 . The engineered immune cell of claim 1 , wherein the engineered immune cell is an induced pluripotent stem cell (iPSC)-derived immune cell.

3 . The engineered immune cell of claim 1 , wherein the engineered immune cell is a peripheral blood (PB)-derived immune cell.

4 . The engineered immune cell of claim 1 , wherein the engineered immune cell is a cord blood (CB)-derived immune cell.

5 . The engineered immune cell of claim 1 , wherein the engineered immune cell is a natural killer (NK) cell.

6 . The engineered immune cell of claim 1 , wherein the ADAM17 inactivating mutation is a knockout of an endogenous ADAM17 gene.

7 . The engineered immune cell of claim 6 , wherein the engineered immune cell is a ADAM17 −/− immune cell.

8 . The engineered immune cell of claim 1 , wherein the ADAM17 inactivating mutation is a knockdown of an endogenous ADAM17 gene.

9 . The engineered immune cell of claim 1 , wherein the engineered immune cell is CD56+, CD94+, NKG2D+, NKp44+, and NKp46+.

10 . The engineered immune cell of claim 1 , wherein the engineered immune cell has increased expression of CD16a, TNF-α, and CD62L (L-selectin) as compared to a non-engineered immune cell after stimulation with a stimulating agent.

11 . The engineered immune cell of claim 1 , wherein expression of CD16a in the engineered immune cell is stably maintained for a period of at least about 6 weeks during expansion in a culture.

12 . The engineered immune cell of claim 1 , wherein the immune cell is a human immune cell.

13 . A purified cell composition comprising one or more of the engineered immune cell of claim 1 .

14 . The purified cell composition of claim 13 , wherein at least about 71% of engineered immune cells express tumor necrosis factor-alpha (TNF-α) after stimulation with a stimulating agent.

15 . The purified cell composition of claim 13 , wherein at least about 42% of engineered immune cells express CD62L (L-selectin) after stimulation with a stimulating agent.

16 . The purified cell composition of claim 13 , wherein the engineered immune cells have increased expression of CD107a and interferon gamma (IFNγ) as compared to non-engineered immune cells after co-incubation with a disease cell line and an antibody specific to said disease cell line.

17 . The purified cell composition of claim 16 , wherein the disease cell line is a B-lymphoma cell line and the antibody specific to said disease cell line is an anti-CD20 antibody.

18 . The purified cell composition of claim 17 , wherein the anti-CD20 antibody is rituximab.

19 . The purified cell composition of claim 17 , wherein the engineered immune cells exhibit at least about 45% increased expression of CD107a.

20 . The purified cell composition of claim 17 , wherein the engineered immune cells exhibit at least about 36% increased expression of IFNγ.

21 . The purified cell composition of claim 16 , wherein the disease cell line is a squamous cell carcinoma cell line and the antibody specific to said disease cell line is an anti-EGFR antibody.

22 . The purified cell composition of claim 21 , wherein the anti-EGFR antibody is cetuximab.

23 . The purified cell composition of claim 21 , wherein the engineered immune cells exhibit at least about 29% increased expression of CD107a.

24 . The purified cell composition of claim 21 , wherein the engineered immune cells exhibit at least about 39% increased expression of IFNγ.

25 . The purified cell composition of claim 13 , wherein the engineered immune cells exhibit enhanced antibody-dependent cellular cytotoxicity (ADCC) as compared to non-engineered immune cells.

26 . A method of making the engineered immune cell of claim 1 comprising:
a) introducing an inactivating mutation in an endogenous A-Disintegrin-And Metalloproteinase 17 (ADAM17) gene into a stem cell; and
b) differentiating the stem cell into an immune cell.

27 . The method of claim 26 , wherein the stem cell is an induced pluripotent stem cell (iPSC), peripheral blood cell, or a cord blood cell.

28 . The method of claim 26 , wherein the immune cell is a natural killer cell (NK) cell.

29 . A pharmaceutical composition comprising the engineered immune cell of claim 1 and one or more pharmaceutically acceptable excipients or diluents.

30 . A kit comprising the engineered immune cell of claim 1 and instructions for use.

31 . A method of treating or preventing a disease or disorder in a subject in need thereof, comprising administering the engineered immune cell of claim 1 to the subject.

32 . The method of claim 31 , wherein the disease or disorder is a malignancy.

33 . The method of claim 32 , wherein the malignancy comprises a tumor-associated antigen.

34 . The method of claim 31 , wherein the disease or disorder is a viral infection.

35 . The method of claim 34 , wherein the viral infection comprises a viral infection-associated antigen.

36 . The method of claim 31 , wherein the administering further comprises administering the engineered immune cell or pharmaceutical composition comprising said engineered immune cell in combination with an antibody specific to a disease.

37 . The method of claim 36 , wherein the antibody specific to a disease is an anti-CD20 antibody.

38 . The method of claim 37 , wherein the anti-CD20 antibody is rituximab.

39 . The method of claim 36 , wherein the antibody specific to a disease is an anti-EGFR antibody.

40 . The method of claim 39 , wherein the anti-EGFR antibody is cetuximab.

41 . A cellular culture comprising genetically modified ADAM17-deficient Natural Killer (NK) cells.

42 . The cellular culture of claim 41 , wherein the NK cells have been produced from induced pluripotent stem cells (iPSCs).

43 . The cellular culture of claim 41 , wherein the NK cells have been produced from peripheral blood cells or cord blood cells.

44 . The cellular culture of claim 41 , wherein the NK cells are human NK cells.

45 . The cellular culture of claim 41 , wherein the NK cells exhibit enhanced antibody-dependent cellular cytotoxicity (ADCC) as compared to non-genetically modified NK cells.

46 . The cellular culture of claim 41 , wherein the NK cells express cell surface markers CD56, NKG2D, NKp44, and NKp46.

47 . The cellular culture of claim 41 , wherein the NK cells express increased cell surface markers CD16, TNF-α, and CD62L as compared to non-genetically modified NK cells.

48 . The cellular culture of claim 41 , wherein the NK cells have been genetically modified to be ADAM17-deficient with CRISPR, TALEN, or ZFN.

49 . A pharmaceutical composition comprising NK cells from the culture of genetically modified ADAM17-deficient NK cells of claim 41 .

50 . A method of treating a subject in need comprising administering to the subject an effective amo

Assignees

• Univ California

Inventors

• Kaufman Dan S
• Yamamoto Kenta

Classifications

• A61K40/42

  Cancer antigens · CPC title

• A61K40/15

  Natural-killer [NK] cells; Natural-killer T [NKT] cells · CPC title

• C07K16/2863 Primary

  against receptors for growth factors, growth regulators · CPC title

• C12N5/0646 Primary

  Natural killers cells [NK], NKT cells · CPC title

• C12N9/6489

  Metalloendopeptidases (3.4.24) · CPC title