Therapeutic cells and methods relating to the same

Abstract

The present disclosure provides cell-based compositions for treating diabetes, as well as methods for preparing such cells.

Description

This disclosure provides cell-based compositions for treating diabetes, and methods for preparing such cells. Abstract

Claims

WHAT IS CLAIMED IS:

1. An isolated synthetic replacement endocrine (SRE) cell that expresses transcription factors comprising:(a) Pdxl and ISL1, or(b) MAFA and ISL1; wherein the SRE cell does not express transcription factor Nkx6.1.

2. The isolated cell of claim 1, wherein the isolated SRE cell is differentiated from a stem cell.

3. The isolated cell of claim 2, wherein the stem cell is selected from an embryonic stem cell, an induced pluripotent stem cell, and a multipotent reprogrammed stem cell.

4. The isolated cell of claim 2, wherein the stem cell is derived from a cell line.

5. The isolated cell of claim 2, wherein the stem cell is a multipotent reprogrammed stem cell.

6. The isolated cell of claim 5, wherein the multipotent reprogrammed stem cell was obtained by reprogramming a pancreatic cell.

7. The isolated cell of claim 6, wherein the multipotent reprogrammed stem cell was reprogrammed with an expression plasmid encoding Oct4, Sox2, Klf4, and L-Myc.

8. The isolated cell of claim 6, wherein the multipotent reprogrammed stem cell was reprogrammed with an expression plasmid encoding Oct4, Sox2, Klf4, and C-Myc.

9. The isolated cell of claim 6, wherein the multipotent reprogrammed stem cell was reprogrammed with an expression plasmid encoding LIN28, Oct4, Sox2, and Nanog.

10. The isolated cell of claim 6, wherein the multipotent reprogrammed stem cell was reprogrammed with an expression plasmid encoding Gilsl, Oct3 / 4, Sox2, and Klf4.

11. The isolated cell of claim 6, wherein the stem cell was genetically modified to avoid targeting by lymphocytes.

12. The isolated cell of claim 6, wherein the stem cell was genetically modified to lack expression of MHCI and MHCII.

13. The isolated cell of claim 6, wherein the stem cell was genetically modified to avoid targeting by natural killer cells.

14. The isolated cell of claim 6, wherein the stem cell was genetically modified to express CD47.

15. The isolated cell of claim 5, wherein the multipotent reprogrammed stem cell can differentiate into endoderm or ectoderm cell types, but not mesoderm cell types.

16. The isolated cell of claim 5, wherein the multipotent reprogrammed stem cell does not comprise any reprogramming genes incorporated into its genome.

17. The isolated cell of claim 1, wherein the isolated SRE cell expresses C-peptide.

18. The isolated cell of claim 17, wherein C-peptide is expressed at levels equivalent to mature, native pancreatic islet cells.

19. The isolated cell of claim 17, wherein C-peptide is expressed at levels higher than an SRE cell that expresses Nkx6.1.

20. The isolated cell of any one of claims 17, wherein C-peptide expression is responsive to fluctuations in glucose concentration.

21. The isolated cell of claim 1, wherein the isolated SRE cell secretes insulin.

22. The isolated cell of one of claim 1, wherein the isolated SRE cell secretes glucagon.

23. The isolated cell of one of claim 1, wherein the isolated SRE cell is human.

24. A pharmaceutical composition, comprising at least one isolated cell according to any one of claims 1-23 and a therapeutically acceptable carrier.

25. A method of treating diabetes, comprising administering to a subject with diabetes the isolated cell accordingly to any one of claims 1-23 or the pharmaceutical composition according to claim 24.

26. The method of claim 25, wherein the diabetes is type 1.

27. The method of claim 25, wherein Nkx6.1 is expressed after the isolated cell or pharmaceutical composition is administered to the subject.

28. The method of claim 25, wherein administration comprising implanting the isolated cell or pharmaceutical composition in the subject.

29. The method of claim 25, wherein the subject is human.

30. A method of preparation of an isolated synthetic replacement endocrine (SRE) cell, comprising: contacting an undifferentiated stem cell with a combination of factors to drive differentiation of the stem cell to an endocrine lineage, wherein the combination of factors comprises retinoic acid, a Hedgehog antagonist, and a bone morphogenetic protein (BMP) signaling inhibitor.

31. The method of claim 30, wherein the Hedgehog antagonist is selected from SANT1 and cyclopamine.

32. The method of claim 30 or 31, wherein the BMP signaling inhibitor is LDN193189.

33. The method of claim 30, wherein the stem cell is not exposed to tri-iodothyronine (T3) or analog thereof during the differentiation process.

34. The method of claim 30, wherein the stem cell is not exposed to a protein kinase C activator during the differentiation process.

35. The method of claim 30, wherein the stem cell is derived from a cell line.

36. The method of claim 30, wherein the stem cell is selected from an embryonic stem cell, an induced pluripotent stem cell, and a multipotent reprogrammed stem cell.

37. The method of claim 30, wherein the stem cell is a multipotent reprogrammed stem cell.

38. The method of claim 37, wherein the multipotent reprogrammed stem cell was obtained by reprogramming a pancreatic cell.

39. The method of claim 38, wherein the multipotent reprogrammed stem cell was reprogrammed with an expression plasmid encoding Oct4, Sox2, Klf4, and L-Myc.

40. The method of claim 38, wherein the multipotent reprogrammed stem cell was reprogrammed with an expression plasmid encoding Oct4, Sox2, Klf4, and C-Myc.41 . The method of claim 38, wherein the multipotent reprogrammed stem cell was reprogrammed with an expression plasmid encoding LIN28, Oct4, Sox2, and Nanog.

42. The method of claim 38, wherein the multipotent reprogrammed stem cell was reprogrammed with an expression plasmid encoding Gilsl, Oct3 / 4, Sox2, and Klf4.

43. The method of any one of claims 37-42, wherein the multipotent reprogrammed stem cell can differentiate into endoderm or ectoderm cell types, but not mesoderm cell types.

44. The method of any one of claims 37-42, wherein the multipotent reprogrammed stem cell does not comprise any reprogramming genes incorporated into its genome.

45. The method of any one of claims 37-42, wherein the stem cell was genetically modified to avoid targeting by lymphocytes.

46. The method of any one of claims 37-42, wherein the stem cell was genetically modified to lack expression of MHCI and MHCII.

47. The method of any one of claims 37-42, wherein the stem cell was genetically modified to avoid targeting by natural killer cells.

48. The method of any one of claims 37-42, wherein the stem cell was genetically modified to express CD47.

49. An isolated synthetic replacement endocrine (SRE) cell obtained by the method of claim 30.

50. The isolated cell of claim 49, wherein the cell expresses transcription factors Pdxl and ISL1, and does not express transcription factor Nkx6.1.

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