Generation and application of universal T cells for B-ALL

• US 20070036773A1
• Filed: 08/08/2006
• Published: 02/15/2007
• Est. Priority Date: 08/09/2005
• Status: Abandoned Application

• Current Assignee: City of Hope
• Original Assignee: City of Hope
• Inventors: Rossi, John, Cooper, Laurence
• Application Number:
  US11/500,490
• Publication Number:
  US 20070036773A1
• Time in Patent Office: Days
• US Class Current: 424/93.210
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• CPC Class Codes: A61K 2239/38 characterised by the dose, timing or administration schedule
• CPC Class Codes: A61K 2239/48 Blood cells, e.g. leukemia or lymphoma
• 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 39/464412 CD19 or B4
• CPC Class Codes: A61K 39/464838 Viral antigens
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: C12N 2501/599 with CD designations not provided for elsewhere
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0636 T lymphocytes

Abstract

The present invention is directed to universal T cells and their use in treating diseases and other physiological conditions. More specifically, the present invention is directed to universal T cells and their use in treating treating B-lineage acute lymphoblastic leukemia (B-ALL) in particular and malignancy in general. The universal T cells contain (i) nucleic acid encoding a chimeric antigen receptor (CAR) to redirect their antigen specificity and effector function and (ii) nucleic acids encoding shRNA and/or siRNA molecules to down-regulate cell-surface expression of T cell classical HLA class I and/or II genes to avoid recognition by recipient T cells. The universal T cells may also contain a nucleic acid encoding a non-classical HLA gene, such as an HLA E gene to enforce expression of HLA E genes and/or an HLA G gene to enforce expression of HLA G genes, to avoid recognition by recipient NK cells. The universal T cells may further contain a nucleic acid encoding a selection-suicide gene.

Claims

1. A genetically engineered T cell comprising stably incorporated in its genome a nucleic acid encoding a chimeric antigen receptor (CAR), one or more nucleic acids each encoding an RNAi molecule corresponding to a gene encoding an HLA class I gene and one or more nucleic acids each encoding an RNAi molecule corresponding to a gene encoding an HLA class II gene.

2. The genetically engineered T cell of claim 1 which further comprises a nucleic acid encoding a non-classical HLA gene stably incorporated in its genome.

3. The genetically engineered T cell of claim 2, wherein the non-classical HLA gene is an HLA E gene.

4. The genetically engineered T cell of claim 1 which further comprises a nucleic acid encoding a selection-suicide protein stably incorporated in its genome.

5. The genetically engineered T cell of claim 2 which further comprises a nucleic acid encoding a selection-suicide protein stably incorporated in its genome.

6. The genetically engineered T cell of claim 3 which further comprises a nucleic acid encoding a selection-suicide protein stably incorporated in its genome.

7. The genetically engineered T cell of claim 1, wherein the CAR is CD19R.

8. The genetically engineered T cell of claim 1, wherein the RNAi molecules corresponding to a gene encoding an HLA class I gene are an shRNA molecule and an siRNA molecule and wherein the RNAi molecules corresponding to a gene encoding an HLA class II gene are an shRNA molecule and an siRNA molecule.

9. The genetically engineered T cell of claim 7, wherein the RNAi molecules corresponding to a gene encoding an HLA class I gene are an shRNA molecule and an siRNA molecule and wherein the RNAi molecules corresponding to a gene encoding an HLA class II gene are an shRNA molecule and an siRNA molecule.

31. A method for treating a disease associated with an antigen comprising administering a therapeutically effective amount of the genetically engineered T cells of claim 1.

32. A method for treating B-lineage acute lymphoblastic leukemia comprising administering a therapeutically effective amount of the genetically engineered T cells of claim 7.

33. A method for treating B-lineage acute lymphoblastic leukemia comprising administering a therapeutically effective amount of the genetically engineered T cells of claim 8.

34. A method for treating a disease associated with an antigen comprising administering a therapeutically effective amount of the genetically engineered T cells of claim 9.

10. A process for making a genetically engineered T cell comprising:

(a) introducing a nucleic acid encoding a chimeric antigen receptor (CAR) into a T cell;

(b) introducing one or more nucleic acids each encoding an RNAi molecule corresponding to a gene encoding an HLA class I gene; and

(c) introducing one or more nucleic acids each encoding an RNAi molecule corresponding to a gene encoding an HLA class II gene.

11. The process of claim 10 which further comprises introducing a nucleic acid encoding a non-classical HLA gene.

12. The process of claim 11, wherein the non-classical HLA gene is an HLA E gene.

13. The process of claim 10 which further comprises introducing a nucleic acid encoding a selection-suicide protein.

14. The process of claim 11 which further comprises introducing a nucleic acid encoding a selection-suicide protein.

15. The process of claim 12 which further comprises introducing a nucleic acid encoding a selection-suicide protein.

16. The process of claim 10, wherein the CAR is CD19R.

17. The process of claim 10, wherein the RNAi molecules corresponding to a gene encoding an HLA class I gene are an shRNA molecule and an siRNA molecule and wherein the RNAi molecules corresponding to a gene encoding an HLA class II gene are an shRNA molecule and an siRNA molecule.

18. The process of claim 16, wherein the RNAi molecules corresponding to a gene encoding an HLA class I gene are an shRNA molecule and an siRNA molecule and wherein the RNAi molecules corresponding to a gene encoding an HLA class II gene are an shRNA molecule and an siRNA molecule.

19. The process of claim 10, wherein the nucleic acids are introduced using a transposon system.

20. The process of claim 19, wherein the transposon system is the sleeping beauty (SB) transposon system.

21. The process of claim 20, wherein the nucleic acids are introduced into the T cells via two vectors and a third vector containing a nucleic acid encoding an SB transposase is also introduced into the T cells.

22. The process of claim 16, wherein the nucleic acids are introduced using a transposon system.

23. The process of claim 22, wherein the transposon system is the sleeping beauty (SB) transposon system.

24. The process of claim 23, wherein the nucleic acids are introduced into the T cells via two vectors and a third vector containing a nucleic acid encoding an SB transposase is also introduced into the T cells.

25. The process of claim 17, wherein the nucleic acids are introduced using a transposon system.

26. The process of claim 25, wherein the transposon system is the sleeping beauty (SB) transposon system.

27. The process of claim 26, wherein the nucleic acids are introduced into the T cells via two vectors and a third vector containing a nucleic acid encoding an SB transposase is also introduced into the T cells.

28. The process of claim 18, wherein the nucleic acids are introduced using a transposon system.

29. The process of claim 28, wherein the transposon system is the sleeping beauty (SB) transposon system.

30. The process of claim 29, wherein the nucleic acids are introduced into the T cells via two vectors and a third vector containing a nucleic acid encoding an SB transposase is also introduced into the T cells.