Method for enhancing bone density or formation

• US 20030158132A1
• Filed: 01/22/2002
• Published: 08/21/2003
• Est. Priority Date: 01/22/2002
• Status: Abandoned Application

• Current Assignee: GenVec, Inc. (Precigen, Inc.)
• Original Assignee: GenVec, Inc. (Precigen, Inc.)
• Inventors: Kovesdi, Imre
• Application Number:
  US10/053,637
• Publication Number:
  US 20030158132A1
• Time in Patent Office: Days
• US Class Current: 514/44
• 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: C07K 14/47 from mammals
• CPC Class Codes: C07K 14/475 Growth factors; Growth regulators
• CPC Class Codes: C07K 14/52 Cytokines; Lymphokines; Interferons
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/02 containing a signal sequence
• CPC Class Codes: C12N 2799/022 where the vector is derived from an adenovirus
• CPC Class Codes: C12N 9/16 acting on ester bonds (3.1)

Abstract

The invention pertains to a method for enhancing bone density or formation. In accordance with the method, a nucleic acid encoding a secreted alkaline phosphatase (SEAP) is administered to a cell in a region of a bone such that the nucleic acid is expressed to produce the SEAP, whereby bone density or formation is enhanced within the region. The method can be employed to produce a bone graft having a cell harboring an exogenous nucleic acid encoding a SEAP. To facilitate the inventive method, the invention provides a recombinant viral vector having a nucleic acid encoding a SEAP. Optionally, a nucleic acid encoding an angiogenic protein and/or a nucleic acid encoding an osteogenic protein is employed in conjunction with the nucleic acid encoding a SEAP.

Claims

1. A method for enhancing bone density or formation, the method comprising administering to at least one first cell associated with a region of a bone a first nucleic acid encoding a secreted alkaline phosphatase (SEAP), such that the first nucleic acid is expressed in the cell to produce the SEAP, whereby bone density or formation is enhanced within the region.

2. The method of claim 1, wherein at least one first nucleic acid is exposed to a cell in vivo in the region of the bone.

3. The method of claim 1, wherein at least one first nucleic acid is exposed to a cell ex vivo, which is then delivered in vivo to the region of the bone.

4. The method of claim 1, further comprising administering to a second cell associated with the region a second nucleic acid encoding an angiogenic protein, such that the second nucleic acid is expressed in the cell to produce the angiogenic protein.

5. The method of claim 4, wherein the angiogenic protein is a vascular endothelial growth factor, a connective tissue growth factor, an angiopoetin, an angiopoetin homologous protein, an angiogenin, an angiogenin-2, or P1 GF.

6. The method of claim 4, wherein the SEAP and the angiogenic protein are a single fusion protein comprising a first SEAP domain and a second angiogenic domain.

7. The method of claim 4, wherein the first cell and the second cell are the same.

8. The method of claim 4, wherein the first nucleic acid and the second nucleic acid are the same.

9. The method of claim 1, further comprising administering to a third cell associated with the region a third nucleic acid encoding an osteogenic protein, such that the third nucleic acid is expressed in the cell to produce the osteogenic protein.

10. The method of claim 9, wherein the osteogenic protein is selected from the group consisting of a bone morphogenic protein, a transforming growth factor, a latent transforming growth factor binding protein, latent membrane protein-1, a heparin-binding neurotrophic factor, growth and differentiation factor-5, a parathyroid hormone, a fibroblast growth factor, an epidermal growth factor, a platelet-derived growth factor, an insulin-like growth factor, a growth factor receptor, a cytokine, a chemotactic factor, a granulocyte/macrophage colony stimulating factor, a LIM mineralization protein, a leukemia inhibitory factor, a hedgehog protein, and midkine.

11. The method of claim 9, wherein the SEAP and the osteogenic protein are a single fusion protein comprising a first SEAP domain and a second osteogenic domain.

12. The method of claim 9, wherein the first cell and the third cell are the same.

13. The method of claim 9, wherein the first nucleic acid and the third nucleic acid are the same.

14. A viral vector comprising at least one first nucleic acid encoding a SEAP.

15. The viral vector of claim 14, which is an adenoviral vector.

16. The viral vector of claim 15, which is deficient in at least one gene function required for viral replication.

17. A bone graft comprising a first cell having a first exogenous nucleic acid encoding a SEAP.

18. The bone graft of claim 17, which further comprises a second cell having second exogenous nucleic acid encoding an angiogenic protein.

19. The bone graft of claim 18, wherein the SEAP and the angiogenic protein are a single fusion protein comprising a first SEAP domain and a second angiogenic domain.

20. The bone graft of claim 18, wherein the angiogenic protein is a vascular endothelial growth factor, a connective tissue growth factor, VEGF2, VEGF-C, a fibroblast growth factor, an angiopoetin, an angiopoetin homologous proteins, an angiogenin, an angiogenin-2, or PI GF.

21. The bone graft of claim 18, wherein the first cell and the second cell are the same.

22. The bone graft of claim 18, wherein the first nucleic acid and the second nucleic acid are the same.

23. The bone graft of claim 17, which further comprises a third cell having a third exogenous nucleic acid encoding an osteogenic protein.

24. The bone graft of claim 23, wherein the SEAP and the osteogenic protein are a single fusion protein comprising a first SEAP domain and a second osteogenic domain.

25. The bone graft of claim 23, wherein the osteogenic protein is selected from the group consisting of a bone morphogenic protein, a transforming growth factor, a latent transforming growth factor binding protein, latent membrane protein-1, a heparin-binding neurotrophic factor, growth and differentiation factor-5, a parathyroid hormone, a fibroblast growth factor, an epidermal growth factor, a platelet-derived growth factor, an insulin-like growth factor, a growth factor receptor, a cytokine, a chemotactic factor, a granulocyte/macrophage colony stimulating factor, a LIM mineralization protein, a leukemia inhibitory factor, a hedgehog protein, and midkine.

26. The bone graft of claim 23, wherein the first cell and the third cell are the same.

27. The bone graft of claim 23, wherein the first nucleic acid and the third nucleic acid are the same.

28. The bone graft of claim 17, which is an allograft or an autograft.

30. The recombinant expression cassette of claim 27, which is a SEAP/MK fusion protein, a SEAP/HBNF fusion protein, or SEAP/VEGF fusion protein.

29. A recombinant expression cassette encoding a fusion protein having a first SEAP domain and a second angiogenic or osteogenic domain.