Compositions and methods for cell dedifferentiation and tissue regeneration

• US 20030212024A1
• Filed: 04/04/2003
• Published: 11/13/2003
• Est. Priority Date: 05/12/2000
• Status: Abandoned Application

• Current Assignee: University of Utah Research Foundation (State of Utah)
• Original Assignee: University of Utah Research Foundation (State of Utah)
• Inventors: Odelberg, Shannon J, Keating, Mark T, Poss, Kenneth D
• Application Number:
  US10/275,828
• Publication Number:
  US 20030212024A1
• Time in Patent Office: Days
• US Class Current: 514/44
• CPC Class Codes: A61K 31/404 Indoles, e.g. pindolol
• CPC Class Codes: A61K 38/1709 from mammals
• CPC Class Codes: A61K 38/1825 Fibroblast growth factor [FGF]
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 41/00 Drugs used in surgical methods, e.g. surgery adjuvants for preventing adhesion or for vitreum substitution
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 2500/80 from animals
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/60 Transcription factors
• CPC Class Codes: C12N 2506/1323 from skeletal muscle cells
• CPC Class Codes: C12N 5/0662 Stem cells

Abstract

The present invention is directed to methods and compositions to induce cellular dedifferentiation and tissue regeneration in vitro and in vivo.

Claims

1. A method of regenerating mammalian tissue, comprising

dedifferentiating differentiated mammalian cells by contacting them with a composition capable of inducing dedifferentiation, regeneration or both,

wherein following dedifferentiation the mammalian cells are capable of proliferating and regenerating into redifferentiated or newly differentiated mammalian cells.

2. The method of claim 1, further comprising subsequently proliferating the dedifferentiated mammalian cells.

3. The method of claim 2, further comprising regenerating mammalian cells, tissue, or organs from the dedifferentiated mammalian cells.

4. The method of claim 1, wherein dedifferentiating is conducted in vivo

5. The method of claim 1, wherein dedifferentiating is conducted ex vivo.

6. The method of claim 1, wherein dedifferentiating comprises contacting the mammalian cells with the composition capable of inducing dedifferentiation for a time sufficient to induce dedifferentiation.

7. The method of claim 1, wherein the dedifferentiating is conducted at the site of an injury.

8. The method of claim 7, wherein the injury is caused by disease or trauma.

9. The method of claim 1, wherein the contacting comprises injecting the composition into the site of injury.

10. The method of claim 1, wherein the contacting comprises injecting the composition systemically.

11. The method of claim 1, wherein the contacting comprises topically applying the composition to the site of injury.

12. The method of claim 1, wherein the contacting comprises implanting a delivery device.

13. The method of claim 1, wherein the mammalian cells are isolated from muscle, skin, bone, joints, eye, lung, heart, vasculature, kidney, pancreas, or nervous tissue.

14. The method of claim 1, wherein the mammalian cells are muscle cells.

15. The method of claim 1, wherein the composition comprises an active polypeptide which is a fibroblast growth factor, a fibroblast growth factor receptor, a bone morphogenic polyp eptide, a bone morphogenic polyp eptide receptor, a Wnt polypeptide, a metalloproteinase polypeptide, msx1, msx2, E2F, frizzled, a SMAD polypeptide or a fatty acid binding polypeptide.

16. The method of claim 15, wherein the active polypeptide is a fusion polypeptide.

17. The method of claim 16, wherein the fusion polypeptide comprises the active polypeptide and a polypeptide that facilitates introduction into said cells.

18. The method of claim 1, wherein the composition comprises a polynucleotide encoding an active polypeptide, wherein the active polypeptide is a fibroblast growth factor, a fibroblast growth factor receptor, a bone morphogenic polypeptide, a bone morphogenic polypeptide receptor, a Wnt polypeptide, a metalloproteinase polypeptide, msx1, msx2, E2F, frizzled, a SMAD polypeptide or a fatty acid binding polypeptide.

19. The method of claim 18, wherein the polynucleotide is operably linked to a promoter.

20. The method of claim 19, wherein the promoter is an inducible promoter.

21. The method of claim 18, wherein the polynucleotide is in a vector.

22. The method of claim 15 or 18, wherein the active polypeptide is msx-1.

23. The method of claim 15 or 18, wherein the active polypeptide is fibroblast growth factor.

24. The method of claim 15 or 18, comprising 2 or more active polypeptides.

25. The method of claim 15 or 18, comprising 3 or more active polypeptides.

35. The invention of claim 1, 26, 27, 32, 33 or 34, wherein the extract is an extract from urodeles, teleost fish, echinoderms, and crustaceans.

36. The method of claim 35, wherein the extract is an extract from a newt.

37. The method of claim 35, wherein the extract is humanized.

26. A composition comprising a carrier and a polypeptide which is a fibroblast growth factor, a fibroblast growth factor receptor, a bone morphogenic polypeptide, a bone morphogenic polypeptide receptor, a Wnt polypeptide, a metalloproteinase polypeptide, msx1, msx2, E2F, frizzled, a SMAD polypeptide or a fatty acid binding polypeptide,

wherein the composition dedifferentiates a mammalian cell.

27. A method, comprising

dedifferentiating differentiated mammalian cells by contacting them with a composition comprising an extract from the regeneration site of an animal such that the composition or extract induces dedifferentiation, regeneration or both,

wherein following dedifferentiation the mammalian cells can proliferate and regenerate into redifferentiated mammalian cells.

28. The method of claim 27, further comprising subsequently proliferating the dedifferentiated mammalian cells.

29. The method of claim 28, further comprising regenerating mammalian cells, a tissue or an organ from the dedifferentiated cells.

30. The method of claim 27, wherein dedifferentiating is conducted in vivo

31. The method of claim 27, wherein dedifferentiating is conducted ex vivo.

32. A composition comprising a carrier and an extract from a regenerating site of an animal, wherein the extract dedifferentiates differentiated mammalian cells.

33. A method of identifying polypeptides that induce dedifferentiation of mammalian cells, comprising:

extracting cells from the regeneration site of an animal,

purifying components of the extract,

applying the purified components to mammalian cells,

observing the amount, if any, of dedifferentiation of the mammalian cells, and

comparing the obtained amount of dedifferentiation to the amount of dedifferentiation achieved by contacting mammalian cells with an extract from a newt regenerating site, wherein about the same or greater dedifferentiating activity indicates the polypeptide is capable of inducing dedifferentiation, regeneration or both.

34. A patch comprising,

a matrix, and

an extract from regenerating site of an animal, wherein the extract dedifferentiates differentiated mammalian cells.

38. A method, comprising dedifferentiating differentiated myotube cells by contacting them with a composition comprising an extract from a regeneration site of newt limbs such that the composition induces dedifferentiation, regeneration or both.

39. The method of 38, wherein said myotube cells are murine.

40. The method of 39, wherein said myotube cells are C2C12 cells.

41. The method of 38, wherein said myotube cells are newt.

42. The method of 38, where in said cells are cultured in vitro.

43. The method of 38, wherein after said dedifferentiation, the myotube cells proliferate.

44. A method, comprising dedifferentiating differentiated myotube cells by contacting said cells with a composition comprising a msx1 polynucleotide.

45. The method of 44, wherein said msx1 polynucleotide is operably-linked to an inducible promoter.

46. The method of 44, wherein said myotube cells are murine.

47. The method of 44, wherein said myotube cells are cultured in vitro.

48. The method of 44, wherein after said dedifferentiation, the myotube cells proliferate.

49. The method of 44, wherein after said dedifferentiation, said cells are pluripotent.

50. A method, comprising inducing blastema formation at an injury site by contacting the injury site with a composition comprising fibroblast growth factor.

51. The method of 50, wherein said fibroblast growth factor is wound fibroblast growth factor.

52. A method comprising inhibiting blastema formation at a site of injury by contacting said site with an inhibitor of fibroblast growth factor receptors.

53. The method of 52, wherein said inhibitor is SU5402.

54. The method of 50 or 52, wherein said injury is in zebrafish.

55. The method of 50 or 52, wherein said injury is incurred by trauma or disease.