Human application of engineered chimeric antigen receptor (CAR) T-cells

Patent metadata

Field	Value
Publication number	US-9629877-B2
Application number	US-201414890977-A
Country	US
Kind code	B2
Filing date	May 14, 2014
Priority date	May 14, 2013
Publication date	Apr 25, 2017
Grant date	Apr 25, 2017

Abstract

The present invention concerns methods and compositions for immunotherapy employing a modified T cell comprising a chimeric antigen receptor (CAR). In particular aspects, CAR-expressing T-cells are producing using electroporation in conjunction with a transposon-based integration system to produce a population of CAR-expressing cells that require minimal ex vivo expansion or that can be directly administered to patients for disease (e.g., cancer) treatment.

First claim

The invention claimed is:

1. An engineered cell comprising:
(a) a chimeric antigen receptor (CAR); and
(b) a fusion protein comprising IL-15 and IL-15Rα.

2. The cell of claim 1 , wherein said fusion protein further comprises a linker connecting said IL-15 and IL-15Rα.

3. The cell of claim 2 , wherein the fusion protein comprises an amino acid sequence which is at least 90% identical to SEQ ID NO:6.

4. The cell of claim 3 , wherein the fusion protein comprises the amino acid sequence of SEQ ID NO:6.

5. The cell of claim 1 , wherein the fusion protein is encoded by a DNA comprising a sequence which is at least 90% identical to SEQ ID NO: 7.

6. The cell of claim 5 , wherein the fusion protein is encoded by a DNA comprising the sequence of SEQ ID NO: 7.

7. The cell of claim 1 , wherein DNA encoding the CAR is integrated into the genome of the cell.

8. The cell of claim 1 , wherein DNA encoding the fusion protein is comprised in an extra chromosomal vector.

9. The cell of claim 1 , wherein DNA encoding the fusion protein is integrated into the genome of the cell.

10. A method of treating cancer in a human subject in need thereof comprising administering to the subject an effective amount of one or more cells in accordance with claim 1 .

11. The method of claim 10 , wherein said cell comprises a CAR that targets a cancer-cell antigen.

12. The method of claim 11 , wherein the subject has undergone a previous anti-cancer therapy.

13. The method of claim 12 , wherein the subject is in remission.

14. The method of claim 12 , wherein the subject is free of symptoms of the cancer but comprises detectable cancer cells.

15. An IL-15, IL-15Rαfusion protein comprising an amino acid sequence which is at least 90% identical to SEQ ID NO: 6.

16. The fusion protein of claim 15 , comprising the amino acid sequence of SEQ ID NO: 6.

17. A polynucleotide comprising a DNA sequence encoding an IL-15, IL-15Rαfusion protein which is at least 90% identical to SEQ ID NO: 6.

18. The polynucleotide of claim 17 comprising a sequence which is at least 90% identical to SEQ ID NO: 7.

19. An engineered cell comprising a polynucleotide in accordance with claim 17 .

20. The cell of claim 19 wherein the cell is a T-cell, a T-cell precursor, a NK cell or an aAPC.

21. The method of claim 11 , wherein the cancer-cell antigen is CD19, ROR1, CD56, EGFR, CD123, c-met, GD2, HER2, CD20, MUC-1, CD23, or CD30.

22. The method of claim 11 , wherein the cancer is leukemia or a solid tumor.

23. The cell of claim 2 , wherein said linker further comprises a serine-glycine linker.

24. The cell of claim 1 , wherein said fusion protein further comprises a IgE signal peptide.

25. An engineered cell comprising DNA encoding:
(a) a chimeric antigen receptor (CAR); and
(b) a fusion protein comprising IL-15 and IL-15Rα.

26. The cell of claim 25 , further comprising a transposase.

27. The cell of claim 26 , wherein the transposase is a Sleeping Beauty transposase.

28. The cell of claim 26 , wherein the transposase is provided as a DNA expression vector.

29. The cell of claim 26 , wherein the transposase is a mRNA encoding a Sleeping Beauty transposase.

30. The cell of claim 29 , wherein the mRNA comprises a cap, a poly-A tail or both.

31. The cell of claim 2 , wherein the fusion protein comprises an amino acid sequence which is at least 85% identical to SEQ ID NO:6.

Assignees

• Univ Texas

Inventors

• Cooper Laurence Jn
• Torikai Hiroki
• Zhang Ling
• Huls Helen
• Wang-Johanning Feng
• Hurton Lenka
• Olivares Simon
• Krishnamurthy Janani

Classifications

• A61P37/00 Primary

  Drugs for immunological or allergic disorders · CPC title

• C07K2319/03

  containing a transmembrane segment · CPC title

• C07K2317/622

  Single chain antibody (scFv) · CPC title

• C07K14/5443

  IL-15 · CPC title

• C07K16/30

  from tumour cells · CPC title