Claims for Patent: 6,312,957

Summary for Patent: 6,312,957

Title:	Genetic modification of primate hemopoietic repopulating stem cells
Abstract:	Genetic modification of pluripotent hemopoietic stem cells of primates (P-PHSC) by transduction of P-PHSC with a recombinant adeno-associated virus (AAV). Tile genomc of the recombinant AAV comprises a DNA sequence flanked by the inverted terminal repeats (ITR) of AAV. The DNA sequence will normally comprise regulatory sequences which are functional in hemopoictic cells and, controlled by these regulatory sequences, a sequence coding for a protein or RNA with a therapeutic property when introduced into hemopoietic cells. Preferred examples of DNA sequences are the human lysosomal glococerebrosidase gene, a globin gene from the human .beta.-globin gene cluster, a DNA sequence encoding an RNA or protein with anti-viral activity, the .alpha.1-antitrypsin gene and the human multidrug resistance gene I (MDRI). The invention provides for effective gene therapy with PHSC of primates, in particular humans.
Inventor(s):	Einerhand; Markus Peter Wilhelmus (Amsterdam, NL), Valerio; Domenico (Leiden, NL)
Assignee:	Introgene B.V. (NL)
Application Number:	09/326,032
Patent Claims:	1. A process of genetically modifying pluripotent hemopoietic stem cells of primates (P-PHSC), said process comprising: harvesting P-PHSC: after said harvesting, culturing said harvested P-PHSC in a culture medium allowing for proliferation of said P-PHSC; and after said culturing, introducing a recombinant adeno-associated virus (AAV) vector into said cultured P-PHSC to genetically modify said cultured P-PHSC. 2. The process of claim 1, wherein the recombinant AAV vector is derived from human AAV. 3. The process of claim 2, wherein the recombinant AAV vector comprises a DNA sequence flanked by AAV inverted terminal repeats (ITR) or by mutant or recombinant ITR sequences that function as an AAV ITR or by fragments of any thereof that function as an ITR. 4. The process of claim 3, wherein said DNA sequence comprises regulatory sequences which are functional in hemopoietic cells and, under control of said regulatory sequences, a sequence coding for a protein or RNA with a therapeutic property when introducedinto hemopoietic cells. 5. The process of claim 4, wherein said DNA sequence comprises a coding sequence of a gene selected from a group consisting of a human lysosomal glucocerebrosidase gene (E.C.3.2.1.45), a globin gene from a human .beta.-globin gene cluster, a DNA sequence encoding an RNA or protein with anti-viral activity, an .alpha.1-antitrypsin gene and a human multidrug resistance gene I (MDRI). 6. The process of claim 5, wherein said DNA sequence comprises the human .beta.-globin gene inclusive of at least one intron. 7. The process of claim 6, wherein said DNA sequence comprises the human .beta.-globin gene operably linked to erythroid-specific DNaseI hypersensitive sites from its Locus Control Region (LCR). 8. The process of claim 7, wherein said erythroid-specific DNaseI hypersensitive sites from .beta.-globin LCR comprise .beta.-LCR elements HS4, HS3 and HS2. 9. The process of claim 8, wherein said DNA sequence comprises the human .beta.-globin gene under transcriptional control of a functional part of a .beta.-globin promoter. 10. The process of claim 9, wherein said DNA sequence comprises a selectable marker gene useful in hemopoietic stem cells. 11. The process of claim 10, wherein said selectable marker gene is a neo.sup.R gene under transcriptional control of a herpes simplex virus (HSV) thymidine kinase (tk) promoter. 12. The process of claim 10, wherein said selectable marker gene is a neo.sup.R gene under transcriptional control of a .DELTA.Mo+PyF101 Long Terminal Repeat (LTR) promoter. 13. The process of claim 11, wherein said recombinant AAV vector is part of a complex when contacted with said P-PHSC. 14. The process of claim 12, wherein said recombinant AAV vector is part of a complex when contacted with said P-PHSC. 15. The process of claim 13, wherein said recombinant AAV vector is associated with AAV capsid proteins. 16. The process of claim 13, wherein said recombinant ector is packaged into an AAV capsid. 17. The process of claim 14, wherein said recombinant AAV vector is associated with AAV capsid proteins. 18. The process of claim 16, wherein said recombinant AAV vector is introduced into said P-PHSC by transduction with the recombinant AAV vector packaged in an AAV capsid. 19. The process of claim 17, wherein said recombinant AAV vector is packaged into an AAV capsid. 20. The process of claim 18, wherein said P-PHSC are obtained from primate bone marrow, cord blood or peripheral blood. 21. The process of claim 19, wherein said recombinant AAV vector is introduced into said P-PHSC by transduction with the recombinant AAV vector packaged in an AAV capsid. 22. The process of claim 20, wherein said P-PHSC are obtained from a human. 23. The process of claim 21 wherein said P-PHSC are obtained from primate bone marrow, cord blood or peripheral blood. 24. The process of claim 22, wherein said P-PHSC are exposed in vitro to one or more proliferation stimulating compounds. 25. The process of claim 23, wherein said P-PHSC are obtained from a human. 26. The process of claim 24, wherein said P-PHSC are exposed in vitro to interleukin 3 or a fragment thereof. 27. The process of claim 25 wherein said P-PHSC are exposed in vitro to one or more proliferation stimulating compounds. 28. The process of claim 27, wherein said P-PHSC are exposed in vitro to interleukin 3 or a fragment thereof.

Details for Patent 6,312,957

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132	06/04/1986		2016-12-05
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132			2016-12-05
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	Injection	103132			2016-12-05