USE OF HEAT SHOCK TO TREAT OCULAR DISEASE

• US 20100068141A1
• Filed: 07/27/2006
• Published: 03/18/2010
• Est. Priority Date: 07/27/2005
• Status: Abandoned Application

• Current Assignee: University of Florida Research Foundation, Incorporated (State University System of Florida)
• Original Assignee: University of Florida (State University System of Florida)
• Inventors: Grant, Maria G., Kaushal, Shalesh
• Application Number:
  US11/989,347
• Publication Number:
  US 20100068141A1
• Time in Patent Office: Days
• US Class Current: 514/1.100
• CPC Class Codes: A61F 2009/00863 Retina
• CPC Class Codes: A61F 2009/00872 Cornea
• CPC Class Codes: A61F 9/008 using laser
• CPC Class Codes: A61K 31/121 acyclic
• CPC Class Codes: A61K 31/167 having the nitrogen of a carboxamide group directly attached to the aromatic ring, e.g. lidocaine, paracetamol
• CPC Class Codes: A61K 31/192 having aromatic groups, e.g. sulindac, 2-aryl-propionic acids, ethacrynic acid cannabinoids A61K31/658
• CPC Class Codes: A61K 31/336 having three-membered rings, e.g. oxirane, fumagillin
• CPC Class Codes: A61K 31/52 Purines, e.g. adenine
• CPC Class Codes: A61K 31/7004 Monosaccharides having only carbon, hydrogen and oxygen atoms
• CPC Class Codes: A61K 38/1709 from mammals
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 27/06 Antiglaucoma agents or miotics
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis

Abstract

The invention generally provides methods for recruiting stem cells to an ocular tissue. The methods involve inducing heat shock in the ocular tissue using a subthreshold laser and/or an agent. In some embodiments, the heat shock is induced following the administration of an agent that mobilizes HSCs.

Claims

1. A method for ameliorating an ocular disease in a subject, the method comprising

(a) inducing heat shock in at least one cell of an ocular tissue;

and

(b) recruiting a stem cell to the ocular tissue, thereby ameliorating the ocular disorder.

2. The method of claim 1, wherein the heat shock is induced in the ocular tissue using sub-visible threshold laser (SVL) stimulation.

3. The method of claim 1, wherein the heat shock is induced using a small compound selected from the group consisting of geldanamycin, celastrol, 17-allylamino-17-demethoxygeldanamycin, EC102, radicicol, geranylgeranylacetone, paeoniflorin, PU-DZ8, and H-71.

4. The method of claim 1, wherein the heat shock is induced using a polypeptide selected from the group consisting of Hsp100, Hsp90, Hsp70, Hsp60, and Hsp40.

5. The method of claim 1, wherein heat shock is induced using an expression vector comprising a polynucleotide encoding a heat shock polypeptide.

6. The method of any one of claims 1-4, wherein the method increases the expression or activity of a heat shock protein selected from the group consisting of Hsp100, Hsp90, Hsp70, Hsp60, and Hsp40.

7. The method of claim 4 or 5, wherein the heat shock polypeptide is Hsp70 or Hsp90.

8. The method of claim 1, wherein the stem cell is a hematopoietic stem cell.

9. The method of claim 1, wherein the method reduces at least one symptom of the ocular disease or disorder.

10. The method of claim 1, wherein the ocular disease or disorder is selected from the group consisting of diabetic retinopathy, choroidal neovascularization, glaucoma retinitis pigmentosa, age-related macular degeneration, glaucoma, corneal dystrophies, retinoschises, Stargardt'"'"'s disease, autosomal dominant druzen, and Best'"'"'s macular dystrophy, cystoid macular edema, retinal detachment, photic damage, ischemic retinopathies, inflammation-induced retinal degenerative disease, X-linked juvenile retinoschisis, glaucoma, Malattia Leventinese (ML) and Doyne honeycomb retinal dystrophy.

19. The method of claim 10, wherein the sub-threshold laser energy is between 10 mW to 100 mW and is administered in a 0.1 msec pulse.

20. The method of claim 10, wherein the stimulation increases the expression or biological activity of a heat shock protein selected from the group consisting of Hsp100, Hsp90, Hsp70, Hsp60, and Hsp40.

21. The method of claim 10, wherein the stimulation alters the expression or activity of a protein selected from the group consisting of SDF-1, VEGF, HIF-1α

, crystallin, hypoxia-inducible factor 1-alpha (HIF-1a), and CXCR-4.

22. The method of claim 10, wherein the method increases the expression of an Hsp70 or Hsp90 polypeptide by at least 10-fold.

23. The method of claim 10, wherein the method increases the expression or activity of an Hsp70 or Hsp90 polypeptide by at least 40-fold.

29. The method of any one of claims 22-28, wherein the heat shock is induced using a small compound.

30. The method of claim 29, wherein the small compound is selected from the group consisting of geldanamycin, celastrol, 17-allylamino-17-demethoxygeldanamycin, EC102, radicicol, geranylgeranylacetone, paeoniflorin, PU-DZ8, and H-71.

31. The method of any one of claims 22-28, wherein the heat shock is induced using a polypeptide.

32. The method of any one of claims 22-28, wherein heat shock is induced using an expression vector comprising a polynucleotide encoding a heat shock polypeptide.

33. The method of claim 30 or 31, wherein the heat shock polypeptide is selected from the group consisting of Hsp100, Hsp90, Hsp70, Hsp60, and Hsp40.

34. The method of any one of claims 22-28, wherein the agent increases the expression or biological activity of a heat shock protein selected from the group consisting of Hsp100, Hsp90, Hsp70, Hsp60, and Hsp40.

35. The method of any one of claims 22-28, wherein the agent alters the expression or activity of a protein selected from the group consisting of SDF-1, VEGF, HIF-1α

, crystallin, hypoxia-inducible factor 1-alpha (HIF-1a), and CXCR-4,

36. The method of any one of claims 22-28, wherein the method increases the expression of an Hsp70 or Hsp90 polypeptide by at least 10-fold.

37. The method of any one of claims 22-28, wherein the method increases the expression or activity of an Hsp70 or Hsp90 polypeptide by at least 40-fold.

38. The method of any one of claims 1-28, wherein an agent that increases hematopoietic stem cell mobilization is administered to the subject prior to induction of the heat shock response.

39. The method of any one of claims 1-28, wherein the method further comprises administering an anti-inflammatory agent or an anti-angiogenic agent.

40. The method of any one of claims 1-28, wherein the method further comprises administering an agent that supports the survival, proliferation, or transdifferentiation of a hematopoietic stem cell.

41. The method of any one of claim 1-28, wherein the subject has an ocular disease or disorder selected from the group consisting of diabetic retinopathy, choroidal neovascularization, glaucoma retinitis pigmentosa, age-related macular degeneration, glaucoma, corneal dystrophies, retinoschises, Stargardt'"'"'s disease, autosomal dominant druzen, and Best'"'"'s macular dystrophy, cystoid macular edema, retinal detachment, photic damage, ischemic retinopathies, inflammation-induced retinal degenerative disease, X-linked juvenile retinoschisis, glaucoma, Malattia Leventinese (ML) and Doyne honeycomb retinal dystrophy.

42. The method of any one of claims 1-28, wherein the method further comprises administering all trans-retinoic acid to enhance the transdifferentiation of the stem cell to a retinal pigment epithelial cell.

51. The method of any one of claims 1-49, wherein the agent is administered by intravitreal or retro-orbital injection.

53. The method of any one of claims 1-49, wherein the method further comprises administering a vector encoding a therapeutic polypeptide.

54. The method of any one of claims 1-49, wherein the method further comprises administering a substantially purified stem cell to the subject.

55. The method of any one of claim 1-49, wherein the stem cell is administered locally by intravitreal or retro-orbital infection.

58. The pharmaceutical composition of claim 54, wherein heat shock polypeptide is selected from the group consisting of Hsp100, Hsp90, Hsp70, Hsp60, and Hsp40.

11. A method of recruiting a stem cell to an ocular tissue of a subject in need thereof, the method comprising stimulating the ocular tissue with a sub-threshold laser, wherein the level of stimulation is sufficient to recruit at least one stem cell to the tissue.

12. The method of claim 11, wherein a 10% or 15% duty cycle is used.

13. The method of claim 11, wherein the sub-threshold laser has a wavelength from at least about 100 nm to 2000 nm.

14. The method of claim 11, wherein the sub-threshold laser energy is from about 5 mW to 200 mW.

15. The method of claim 11, wherein the sub-threshold laser energy is from 10 mW to 100 mW.

16. The method of claim 11, wherein the laser is administered in a micropulse.

17. The method of claim 11, wherein the duration of the micropulse is from about 0.001 msec to 1.0 msec.

18. The method of claim 11, wherein the duration of the micropulse is 0.1 msec.

24. A method of recruiting a stem cell to an ocular tissue of a subject in need thereof, the method comprising

(a) administering an agent to a subject in an amount sufficient to induce heat shock in an ocular tissue;

and

(b) recruiting a stem cell to the ocular tissue.

25. A method of ameliorating an ocular disease or disorder in a subject in need thereof, the method comprising

(a) administering an agent to a subject in an amount sufficient to induce heat shock in an ocular tissue;

and

(b) recruiting a stem cell to the ocular tissue, thereby ameliorating the ocular disease or disorder.

26. The method of claim 25, wherein the ocular disease or disorder is selected from the group consisting of diabetic retinopathy, choroidal neovascularization, glaucoma retinitis pigmentosa, age-related macular degeneration, glaucoma, corneal dystrophies, retinoschises, Stargardt'"'"'s disease, autosomal dominant druzen, and Best'"'"'s macular dystrophy, cystoid macular edema, retinal detachment, photic damage, ischemic retinopathies, inflammation-induced retinal degenerative disease, X-linked juvenile retinoschisis, glaucoma, Malattia Leventinese (ML) and Doyne honeycomb retinal dystrophy.

27. A method of regenerating the retina in a subject in need thereof, the method comprising

(a) administering an agent to a subject in an amount sufficient to induce heat shock in an ocular tissue;

and

(b) recruiting a stem cell to the ocular tissue, thereby regenerating the retina.

28. A method of repairing retinal pigment epithelium damage in a subject in need thereof, the method comprising

(a) administering an agent to a subject in an amount sufficient to induce heat shock in an ocular tissue;

and

(b) recruiting a stem cell to the ocular tissue, thereby repairing the retinal pigment epithelium.

43. A method of ameliorating an ocular disease or disorder in a subject in need thereof, the method comprising

(a) administering to the subject an agent that mobilizes a bone marrow derived stem cell in the subject;

(b) inducing heat shock in an ocular tissue; and

(c) recruiting the stem cell to the ocular tissue, thereby ameliorating the ocular disease or disorder.

44. The method of claim 43, wherein the agent is granulocyte macrophage colony stimulating factor or stem cell factor.

45. The method of claim 43, wherein the heat shock is induced using a subthreshold laser treatment.

46. The method of claim 43, wherein the heat shock is induced using an agent that is a small compound, polypeptide, or nucleic acid molecule.

47. The method of claim 45, wherein the small compound is selected from the group consisting of geldanamycin, celastrol, 17-allylamino-17-demethoxygeldanamycin, EC 102, radicicol, geranylgeranylacetone, paeoniflorin, PU-DZ8, and H-71.

48. The method of claim 43, wherein the ocular disease or disorder is selected from the group consisting of diabetic retinopathy, choroidal neovascularization, glaucoma retinitis pigmentosa, age-related macular degeneration, glaucoma, corneal dystrophies, retinoschises, Stargardt'"'"'s disease, autosomal dominant druzen, and Best'"'"'s macular dystrophy, cystoid macular edema, retinal detachment, photic damage, ischemic retinopathies, inflammation-induced retinal degenerative disease, X-linked juvenile retinoschisis, glaucoma, Malattia Leventinese (ML) and Doyne honeycomb retinal dystrophy.

49. A method of ameliorating macular degeneration in a subject in need thereof, the method comprising

(a) administering to the subject GM-CSF and/or Stem Cell Factor, wherein the administration mobilizes a bone marrow derived stem cell in the subject;

(b) inducing heat shock in an ocular tissue by administering a subthreshold laser treatment or agent; and

(c) recruiting the bone marrow derived stem cell to the ocular tissue, thereby ameliorating the macular degeneration.

50. The method of claim 49, wherein the agent is selected from the group consisting of geldanamycin, celastrol, 17-allylamino-17-demethoxygeldanamycin, EC102, radicicol, geranylgeranylacetone, paeoniflorin, PU-DZ8, and H-71.

52. The method of claim 49, wherein the administration induces cellular repair of the RPE layer.

56. A pharmaceutical composition for stem cell recruitment, the composition comprising an effective amount of a small compound selected from the group consisting of geldanamycin, celastrol, 17-allylamino-17-demethoxygeldanamycin, EC102, radicicol, geranylgeranylacetone, paeoniflorin, PU-DZ8, and H-71 in a pharmaceutically acceptable excipient, the composition formulated for ocular delivery.

57. A pharmaceutical composition for stem cell recruitment in an ocular tissue, the composition comprising an expression vector comprising a polynucleotide encoding a heat shock polypeptide in a pharmaceutically acceptable excipient, the composition formulated for ocular delivery.

59. A pharmaceutical composition for stem cell recruitment in an ocular tissue, the composition comprising a polypeptide selected from the group consisting of Hsp100, Hsp90, Hsp70, Hsp60, and Hsp40 in a pharmaceutically acceptable excipient.

60. A kit comprising an effective amount of an agent that induces a heat shock response in an ocular tissue, and instructions for using the kit to increase stem cell recruitment.

61. The kit of claim 60, wherein the agent is a polypeptide, a polynucleotide, or a small compound.

62. The kit of claim 60, wherein the polypeptide is Hsp100, Hsp90, Hsp70, Hsp60, and Hsp40.

63. The kit of claim 60, wherein the polynucleotide encodes Hsp100, Hsp90, Hsp70, Hsp60, and Hsp40.

64. The kit of claim 60, wherein the small compound is selected from the group consisting of geldanamycin, celastrol, 17-allylamino-17-demethoxygeidanamycin, EC102, radicicol, geranylgeranylacetone, paeoniflorin, PU-DZ8, and H-71.

65. A method of identifying an agent that increases stem cell recruitment in an ocular tissue, the method comprising:

(a) contacting an ocular cell with a test compound;

(b) identifying an increase in the expression or activity of a heat shock polypeptide relative to an untreated ocular cell, thereby identifying a compound that increases stem cell recruitment.

66. A method of identifying an agent that increases stem cell recruitment in an ocular tissue, the method comprising:

(a) contacting an ocular cell with a test compound;

(b) identifying an increase in the number of stem cells in the tissue.