Differentiation of non-insulin producing cells into insulin producing cells by GLP-1 or exendin-4 and uses thereof

• US 7,056,734 B1
• Filed: 08/10/1999
• Issued: 06/06/2006
• Est. Priority Date: 08/10/1998
• Status: Expired due to Fees

• Current Assignee: The General Hospital Corporation (Mass General Brigham, Inc.), The United States of America As Represented By The Secretary of Agriculture
• Original Assignee: The United States of America As Represented By The Secretary of Agriculture
• Inventors: Greig, Nigel, Holloway, Harold, Stoffers, Doris, Egan, Josephine, Passaniti, Antonino, Habener, Joel, Perfetti, Riccardo
• Primary Examiner(s): Spector, Lorraine
• Assistant Examiner(s): Jiang, Dong
• Application Number:
  US09/762,538
• Time in Patent Office: 2,492 Days
• Field of Search: 435/325, 424/93.7, 514/2, 514/3
• US Class Current: 435/325
• CPC Class Codes: A61K 35/39 Pancreas; Islets of Langerhans Langerhans cells of epidermis A61K35/36
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 38/26 Glucagons
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 5/48 of the pancreatic hormones
• CPC Class Codes: A61P 5/50 for increasing or potentiating the activity of insulin
• CPC Class Codes: C07K 14/57563 Vasoactive intestinal peptide [VIP]; Related peptides
• CPC Class Codes: C12N 2501/335 Glucagon; Glucagon-like peptide [GLP]; Exendin
• CPC Class Codes: C12N 5/0676 Pancreatic cells

Abstract

The present invention relates to a population of insulin producing cells made by a process comprising contacting non-insulin producing cells with a growth factor selected from the group consisting of GLP-1 or Exendin-4, growth factors having amino acid sequences substantially homologous to GLP-1 or Exendin-4, and fragmets thereof. The present invention also relates to methods of differentiating non-insulin producing cells into insulin producing cells and of enriching a population of cells for insulin-producing cells. The present invention also relates to methods of treating diabetes.

Claims

1. An isolated population of insulin-producing cells obtained from non-insulin-producing cells by a process comprising contacting the non-insulin-producing cells in vitro for at least twenty-four hours with an amount of a substance effective to induce insulin production, wherein the substance is selected from the group consisting of a GLP-1 peptide, a GLP-1 peptide containing one or more conservative amino acid substitutions at positions other than positions 7, 10, 12, 13 and 15 of GLP-1, and a fragment of any one of the preceding GLP-1 peptides, and wherein the GLP-1 peptide or fragment thereof has the ability to differentiate non-insulin-producing cells into insulin-producing cells.

2. The population of claim 1, wherein the non-insulin-producing cells comprise pancreatic cells.

3. The population of claim 1, wherein the non-insulin-producing cells comprise pancreatic acinar cells.

4. The population of claim 1, wherein the non-insulin-producing cells comprise stem cells.

5. The population of claim 1, wherein the non-insulin-producing cells comprise pancreatic stem cells.

6. The population of claim 1, wherein the non-insulin-producing cells comprise mammalian cells.

7. The population of claim 6, wherein the mammalian cells comprise human cells.

8. The composition of claim 1, wherein the contacting is at least 3 days.

9. The composition of claim 1, wherein the substance is a GLP-1 peptide.

10. The composition of claim 1, wherein the substance is a GLP-1 peptide containing one or more conservative amino acid substitutions at positions other than positions 7, 10, 12, 13 and 15 of GLP-1.

11. The composition of claim 1, wherein the substance is the fragment of a GLP-1 peptide, or a GLP-1 peptide containing one or more conservative amino acid substitutions at positions other than positions 7, 10, 12, 13 and 15 of GLP-1.

12. A method for differentiating non-insulin-producing cells into insulin-producing cells, comprising contacting the non-insulin-producing cells in vitro for at least twenty four hours with an amount of a substance effective to induce differentiation of non-insulin-producing cells into insulin-producing cells, wherein the substance is selected from the group consisting of a GLP-1 peptide, a GLP-1 peptide containing one or more conservative amino acid substitutions at positions other than positions 7, 10, 12, 13 and 15 of GLP-1, and a fragment of any one of the preceding GLP-1 peptides, and wherein the GLP-1 peptide or fragment thereof has the ability to differentiate non-insulin-producing cells into insulin-producing cells.

13. The method of claim 12, wherein the non-insulin-producing cells comprise pancreatic cells, pancreatic acinar cells, stem cells, pancreatic stem cells, or mammalian cells.

14. The method of claim 13, wherein the mammalian cells are human cells.

15. The method of claim 12, wherein the contacting is at least 3 days.

16. The method of claim 12, wherein the substance is a GLP-1 peptide.

17. The method of claims 12, wherein the substance is a GLP-1 peptide containing one or more conservative amino acid substitutions at positions other than positions 7, 10, 12, 13 and 15 of GLP-1.

18. The method of claims 12, wherein the substance is the fragment of a GLP-1 peptide, or a GLP-1 peptide containing one or more conservative amino acid substitutions at positions other than positions 7, 10, 12, 13 and 15 of GLP-1.

19. A method of enriching an isolated population of cells for insulin-producing cells, comprising contacting non-insulin-producing cells in vitro for at least twenty four hours with an amount of a substance effective to induce differentiation of non-insulin-producing cells into insulin-producing cells, wherein the substance is selected from the group consisting of a GLP-1 peptide, a GLP-1 peptide containing one or more conservative amino acid substitutions at positions other than positions 7, 10, 12, 13 and 15 of GLP-1, and a fragments of any one of the preceding peptides, and wherein the peptide or fragments thereof has the ability to differentiate non-insulin-producing cells into insulin-producing cells.

20. A method of promoting pancreatic amylase-producing cells to produce insulin, comprising contacting the pancreatic amylase-producing cells in vitro for at least twenty-four hours with an amount of a substance effective to induce insulin production, wherein the substance is selected from the group consisting of a GLP-1 peptide, a GLP-1 peptide containing one or more conservative amino acid substitutions at positions other than positions 7, 10, 12, 13 and 15 of GLP-1, and a fragment of any one of the preceding peptides, and wherein the GLP-1 peptide or fragment thereof has the ability to differentiate non-insulin-producing cells into insulin-producing cells.

21. An isolated population of insulin-producing cells obtained from non-insulin-producing cells by a process comprising contacting the non-insulin-producing cells in vitro for at least twenty-four hours with an amount of a substance effective to induce insulin production, wherein the substance is selected from the group consisting of an Exendin-4 peptide, an Exendin-4 peptide containing one or more conservative amino acid substitutions at positions other than positions 1, 4, 6, 7 and 9 of Exendin-4, and a fragment of any one of the preceding Exendin-4 peptides, and wherein the Exendin-4 peptide or fragment thereof has the ability to differentiate non-insulin-producing cells into insulin-producing cells.

22. The population of claim 21, wherein the non-insulin-producing cells comprise pancreatic cells.

23. The population of claim 21, wherein the non-insulin-producing cells comprise pancreatic acinar cells.

24. The population of claim 21, wherein the non-insulin-producing cells comprise stem cells.

25. The population of claim 21, wherein the non-insulin-producing cells comprise pancreatic stem cells.

26. The population of claim 21, wherein the non-insulin-producing cells comprise mammalian cells.

27. The population of claim 26, wherein the mammalian cells comprise human cells.

28. The composition of claim 21, wherein the contacting is at least 3 days.

29. The composition of claim 21, wherein the substance is exendin-4.

30. The composition of claim 21, wherein the substance is an Exendin-4 peptide containing one or more conservative amino acid substitutions at positions other than positions 1, 4, 6, 7 and 9 of Exendin-4.

31. The composition of claim 21, wherein the substance is the fragment of an Exendin-4 peptide, or an Exendin-4 peptide containing one or more conservative amino acid substitutions at positions other than 1, 4, 6, 7 and 9 of Exendin-4.

32. A method for differentiating non-insulin-producing cells into insulin-producing cells, comprising contacting the non-insulin-producing cells in vitro for at least twenty four hours with an amount of a substance effective to induce differentiation of non-insulin-producing cells into insulin-producing cells, wherein the substance is selected from the group consisting of an Exendin-4 peptide, an Exendin-4 peptide containing one or more conservative amino acid substitutions at positions other than positions 1, 4, 6, 7 and 9 of Exendin-4, and a fragment of any one of the preceding Exendin-4 peptides, and wherein the Exendin-4 peptide or fragment thereof has the ability to differentiate non-insulin-producing cells into insulin-producing cells.

33. The method of claim 32, wherein the non-insulin-producing cells comprise pancreatic cells, pancreatic acinar cells, stem cells, pancreatic stem cells, or mammalian cells.

34. The method of claim 33, wherein the mammalian cells are human cells.

35. The method of claim 32, wherein the contacting is at least 3 days.

36. The method of claim 32, wherein the substance is exendin-4.

37. The method of claim 32, wherein the substance is an Exendin-4 peptide containing one or more conservative amino acid substitutions at positions other than positions 1, 4, 6, 7 and 9 of Exendin-4.

38. The method of claims 32, wherein the substance is the fragment of an Exendin-4 peptide, or an Exendin-4 peptide containing one or more conservative amino acid substitutions at positions other than 1, 4, 6, 7 and 9 of Exendin-4.

39. A method of promoting pancreatic amylase-producing cells to produce insulin, comprising contacting the pancreatic amylase-producing cells in vitro for at least twenty-four hours with an amount of a substance effective to induce insulin production, wherein the substance is selected from the group consisting of an Exendin-4 peptide, an Exendin-4 peptide containing one or more conservative amino acid substitutions at positions other than 1, 4, 6, 7 and 9 of Exendin-4, and a fragment of any one of the preceding peptides, and wherein the Exendin-4 peptide or fragment thereof has the ability to differentiate non-insulin-producing cells into insulin-producing cells.

40. A method of enriching an isolated population of cells for insulin-producing cells, comprising contacting non-insulin-producing cells in vitro for at least twenty-four hours with an amount of a substance effective to induce differentiation of non-insulin-producing cells into insulin-producing cells, wherein the substance is selected from the group consisting of an Exendin-4 peptide, an Exendin-4 peptide containing one or more conservative amino acid substitutions at positions other than 1, 4, 6, 7 and 9 of Exendin-4, and a fragment of any one of the preceding peptides, and wherein the peptide or fragment thereof has the ability to differentiate non-insulin-producing cells into insulin-producing cells.