Methods for promoting wound healing and treating transplant-associated vasculopathy

• US 6,103,230 A
• Filed: 10/02/1998
• Issued: 08/15/2000
• Est. Priority Date: 11/25/1992
• Status: Expired due to Fees

• Current Assignee: University of Pittsburgh of The Commonwealth System of Higher Education
• Original Assignee: University of Pittsburgh of The Commonwealth System of Higher Education
• Inventors: Billiar, Timothy R., Shears, Larry L. II, Tzeng, Edith, Edington, Howard David James, Geller, David A.
• Primary Examiner(s): Achutamurthy, Ponnathapu
• Assistant Examiner(s): Moore, William W
• Application Number:
  US09/165,882
• Time in Patent Office: 683 Days
• Field of Search: 424/94.1, 424/94.4, 435/187
• US Class Current: 424/94.4
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/519 ortho- or peri-condensed with heterocyclic rings
• CPC Class Codes: A61K 38/44 Oxidoreductases (1)
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2710/10343 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 2800/108 episomal vectors
• CPC Class Codes: C12N 2830/42 being an intron or intervening sequence for splicing and/or stability of RNA
• CPC Class Codes: C12N 2840/20 translation of more than one cistron
• CPC Class Codes: C12N 2840/203 having an IRES
• CPC Class Codes: C12N 9/0022 with oxygen as acceptor (1.4.3)
• CPC Class Codes: C12N 9/0071 acting on paired donors with incorporation of molecular oxygen (1.14)
• CPC Class Codes: C12N 9/0075 Nitric-oxide synthase (1.14.13.39)
• CPC Class Codes: C12Y 114/13039 Nitric-oxide synthase (NADPH dependent) (1.14.13.39)

Abstract

The present invention provides a method of promoting the closure of a wound in a patient. This method involves transferring exogenous iNOS to the region of the wound whereby a product of iNOS is produced in the region of the wound to promote the closure of the wound. The present invention also provides a method of transplanting a graft into a patient, which method involves transferring a vector comprising an iNOS expression cassette to cells associated with the graft and surgically incorporating the graft tissue into the patient. Upon expression of iNOS within the patient, a product of iNOS is produced in the region of the graft to attenuate vasculopathy in the region of the graft.

Claims

1. A method of promoting the closure of a wound in a patient which comprises transferring exogenous iNOS to the region of said wound by transferring to cells associated with said wound a vector comprising an iNOS expression cassette, such that iNOS is produced in said cells, whereby a product of iNOS is produced in the region of said wound to promote the closure of said wound.

2. The method of claim 1, wherein said cells are cells of said wound and said vector is transferred to said cells in situ.

3. The method of claim 1, wherein said cells are not the cells of said wound.

4. The method of claim 3, wherein said cells are within a graft and said vector is transferred to said cells ex vivo.

5. The method of claim 3, wherein said cells are cells in vitro, and the cells are transferred to the region of the wound.

6. The method of claim 1, wherein said patient is human.

7. The method of claim 1, wherein said patient is deficient for NO production.

8. The method of claim 1, which further comprises transferring exogenous BH₄ to the region of said wound such that iNOS enzymatic activity is enhanced.

9. The method of claim 8, wherein said wound is an external wound and wherein exogenous BH₄ is transferred to the region of said wound by topical administration of BH₄ to the region of the wound.

10. The method of claim 1, which further comprises transferring to cells associated with said wound a vector comprising a GTPCH expression cassette such that said cells produce BH₄.

11. A method of transplanting a graft into a patient comprising:

a) transferring to cells associated with said graft a vector, said vector comprising an iNOS expression cassette, and

b) surgically incorporating said graft tissue into said patient such that said graft survives within said patient,

whereby iNOS is expressed in said cells in the region of said graft within said patient, and

whereby a product of iNOS is produced in the region of said graft to attenuate vasculopathy in the region of said graft.

12. The method of claim 11, whereby said product of iNOS attenuates vasculopathy by inhibiting neointimal hyperplasia.

13. The method of claim 11, wherein said cells associated with said graft are cells within the tissue comprising said graft.

14. The method of claim 11, wherein said cells associated with said graft are cells of the patient in the region of said graft.

15. The method of claim 11, wherein said cells are cells in vitro which are transferred to the region of said graft.

16. The method of claim 11, wherein said graft comprises vasculature.

17. The method of claim 11, which further comprises transferring to cells associated with said graft a vector comprising a GTPCH expression cassette.

18. The method of claim 11, wherein said vector is transferred to the region of said graft prior to surgically incorporating said graft tissue into said patient.

19. The method of claim 11, wherein said vector is transferred to the region of said graft subsequent to surgically incorporating said graft tissue into said patient.

20. The method of claim 11, wherein said patient is human.