Gene therapy to promote angiogenesis

Abstract

A nucleic acid delivery vehicle for enhancing and/or inducing angiogenesis. This nucleic acid delivery vehicle includes a nucleic acid having at least one sequence coding for a protein capable of increasing nitric oxide production and a nucleic acid delivery carrier. The vehicle can be used in a method for enhancing and/or inducing angiogenesis in an individual involving providing cells of the individual with the nucleic acid delivery vehicle. Also disclosed is a cell for producing the nucleic acid delivery vehicle for enhancing and/or inducing angiogenesis. As before, the nucleic acid delivery vehicle includes a nucleic acid having at least one sequence coding for a protein capable of increasing nitric oxide production and a nucleic acid delivery carrier wherein the nucleic acid delivery carrier includes a virus vector selected from the group of adenoviral vectors and adeno-associated viral vectors, the cell comprising means for producing the virus vector in the absence of replication competent adenovirus and adeno-associated virus.

What is claimed is:  
     
         1 . A nucleic acid delivery vehicle for enhancing and/or inducing angiogenesis, said nucleic acid delivery vehicle comprising a nucleic acid comprising at least one sequence coding for a protein capable of increasing nitric oxide production, and further comprising a nucleic acid delivery carrier.  
     
     
         2 . The nucleic acid delivery vehicle of  claim 1 , wherein said at least one sequence codes for a nitric oxide synthetase or a functional analogue or a derivative thereof.  
     
     
         3 . The nucleic acid delivery vehicle of  claim 1  or  claim 2 , further comprising at least one sequence encoding an additional angiogenesis promoting factor.  
     
     
         4 . The nucleic acid delivery vehicle of  claim 3 , wherein said additional angiogenesis promoting factor is VGEF, bFGF, angiopoietin-1, or functional analogues or derivatives thereof.  
     
     
         5 . The nucleic acid delivery vehicle of any one of claims  1 - 4 , wherein the expression of at least one sequence is regulated by a signal.  
     
     
         6 . The nucleic acid delivery vehicle of  claim 5 , wherein said signal is provided by the oxygen tension in a cell.  
     
     
         7 . The nucleic acid delivery vehicle of  claim 5  or  claim 6 , wherein at least one sequence is expressed from a hypoxia inducible factor 1α promoter.  
     
     
         8 . The nucleic acid delivery vehicle of any one of claims  1 - 7 , further comprising a sequence encoding a herpes simplex virus thymidine kinase or a functional analogue or a derivative thereof.  
     
     
         9 . The nucleic acid delivery vehicle of any one of claims  1 - 8 , wherein said nucleic acid delivery carrier comprises an adenovirus vector or an adeno-associated virus vector.  
     
     
         10 . The nucleic acid delivery vehicle of any one of claims  1 - 9 , wherein said nucleic acid delivery vehicle has at least a partial tissue tropism for muscle cells.  
     
     
         11 . The nucleic acid delivery vehicle of any one of claims  1 - 10 , wherein said nucleic acid delivery vehicle has been at least in part deprived of a tissue tropism for liver cells.  
     
     
         12 . The nucleic acid delivery vehicle of  claim 10  or  claim 11 , wherein said tissue tropism is provided at least in part through a tissue tropism determining part of fiber protein of a subgroup B adenovirus.  
     
     
         13 . The nucleic acid delivery vehicle of  claim 12 , wherein said subgroup B adenovirus is adenovirus  16 .  
     
     
         14 . A method for enhancing and/or inducing angiogenesis in an individual comprising providing cells of the individual with The nucleic acid delivery vehicle of any one of claims  1 - 13 .  
     
     
         15 . The method according to  claim 14  for enhancing and/or inducing angiogenesis with at least one additional angiogenesis promoting factor or parts or derivatives or functional analogues thereof.  
     
     
         16 . The method according to  claim 14  or  claim 15 , wherein the effect of enhancing and/or inducing angiogenesis is at least in part reversible.  
     
     
         17 . The method according to  claim 16 , wherein said effect is at least in part reversed though an increase in the oxygen tension or through providing said cells with gancyclovir or a functional analogue thereof, or both.  
     
     
         18 . The method according to any one of claims  14 - 17 , wherein said cells of the individual comprise at least cells that under normal circumstances are not in direct contact with blood.  
     
     
         19 . The method according to  claim 18 , wherein said cells of the individual are muscle cells.  
     
     
         20 . The method according to  claim 18  or  claim 19  wherein said cells of the individual are smooth muscle cells.  
     
     
         21 . A method of treating endothelial dysfunction in a subject, said method comprising administering the nucleic acid delivery vehicle of any one of claims  1 - 13  to the subject.  
     
     
         22 . A cell for the production of a nucleic acid delivery vehicle of any one of claims  9 - 13  for enhancing and/or inducing angiogenesis, the nucleic acid delivery vehicle comprising a nucleic acid comprising at least one sequence coding for a protein capable of increasing nitric oxide production, and further comprising a nucleic acid delivery carrier wherein said nucleic acid delivery carrier comprises a virus vector selected from the group of an adenovirus vector and an adeno-associated virus vector, said cell comprising means for the production of said virus vector in the absence of replication competent adenovirus and adeno-associated virus.  
     
     
         23 . The cell of  claim 22 , wherein said cell expresses at least one means for the production of said virus vector from a nucleic acid integrated in the chromosomal DNA of said cell and expresses other means for the production of said virus vector from nucleic acid not integrated in the chromosomal DNA of said cell and wherein said integrated nucleic acid and said non-integrated nucleic acid do not comprise sequence overlap leading to the formation of replication competent adenovirus.  
     
     
         24 . The cell of  claim 23 , wherein said integrated nucleic acid comprises at least an adenovirus E1-region or a functional analogue or a derivative thereof.  
     
     
         25 . The cell of  claim 23  or  claim 24 , wherein said integrated nucleic acid comprises at least a sequence encoding an adenovirus E2A protein, preferably an E2A-protein derived from adenovirus ts125 or functional analogues or derivatives thereof.  
     
     
         26 . The cell of any one of claims  22 - 25 , wherein said integrated nucleic acid comprises at least an adenovirus E4-region or a functional analogue or a derivative thereof.  
     
     
         27 . The cell of any one of claims  22 - 26 , wherein said cell is derived from a PER.C6 cell (ECACC deposit number 96022940).