Polypeptides and polynucleotides encoding same and use thereof in the treatment of medical conditions associated with ischemia

Abstract

An isolated polynucleotide is disclosed comprising a nucleic acid sequence encoding a polypeptide having an amino acid sequence of HIF-1 alpha, the polypeptide being stably expressed and constitutively active. Isolated polypeptides encoded by same are also disclosed and uses thereof.

1 . A method of treating a medical condition associated with hypoxia or ischemia, reduced tissue perfusion inclusion or low flow', the method comprising administering to a subject in need thereof a therapeutically effective amount of a nucleic acid construct comprising:
 a) an isolated polynucleotide comprising a nucleic acid sequence encoding HIF-1α polypeptide having an amino acid sequence at least 90% identical to SEQ ID NO: 3 and comprising a mutation at a position corresponding to proline 402 of SEQ ID NO: 3, a mutation at a position corresponding to proline at 564 of SEQ ID NO: 3 and a mutation at a position corresponding to asparagine 803; and   b) an endothelial specific promoter element linked to said isolated polynucleotide, such that said polypeptide is stably expressed and constitutively active in endothelial cells in an endothelial cell specific manner, thereby treating said medical condition in said subject.   
     
     
         2 . The method of  claim 1 , wherein said a mutation at a position corresponding to proline 402 of SEQ ID NO: 3 is a mutation to Alanine. 
     
     
         3 . The method of  claim 1 , wherein said mutation at a position corresponding to proline at 564 of SEQ ID NO: 3 is a mutation to Glycine. 
     
     
         4 . The method of  claim 1 , wherein said mutation at a position corresponding to asparagine 803 is a mutation to Alanine. 
     
     
         5 . The method of  claim 1 , wherein said mutation at a position corresponding to proline 402 of SEQ ID NO: 3 is a mutation to Alanine, wherein said mutation at a position corresponding to proline at 564 of SEQ ID NO: 3 is a mutation to Glycine and wherein said mutation at a position corresponding to asparagine 803 is a mutation to Alanine. 
     
     
         6 . The method of  claim 1 , wherein said HIF-1αpolypeptide comprises the amino acid sequence as set forth in SEQ ID NO: 2. 
     
     
         7 . The method of  claim 1 , wherein said isolated polynucleotide comprises the nucleic acid sequence as set forth in SEQ ID NO: 1. 
     
     
         8 . The method of  claim 1 , wherein said endothelial specific promoter element comprises a PPE-1 promoter. 
     
     
         9 . The method of  claim 8 , wherein said endothelial specific promoter is a PPE-1-3× promoter, as disclosed in PCT/IL01/01059. 
     
     
         10 . The method of  claim 1 , wherein said nucleic acid construct further comprises a hypoxia response element as set forth in SEQ ID NO: 5. 
     
     
         11 . The method of  claim 1 , wherein said medical condition associated with ischemia is selected from the group consisting of wound healing, ischemic stroke, ischemic heart disease, peripheral vascular disease, renal artery disease, gastrointestinal lesions, burns, skin transplantation, vascular grafts, organ repair, bone reparative disorders, liver disorders, uterine disorders, ocular angiogenesis disorders, bone regeneration disorders, cartilage repair disorders and smooth muscle cell disorders. 
     
     
         12 . The method of  claim 1 , wherein said medical condition associated with ischemia is retinal detachment. 
     
     
         13 . The method of  claim 1 , wherein said medical condition associated with ischemia is age related macular degeneration. 
     
     
         14 . The method of  claim 1 , wherein said nucleic acid construct is administered systemically to said subject in need thereof. 
     
     
         15 . The method of  claim 14 , wherein said systemic administration is selected from the group consisting of subcutaneous, intramedullary, intrathecal, direct intraventricular, intravenous, intraperitoneal, intranasal, or intraocular injection. 
     
     
         16 . The method of  claim 1 , wherein said isolated polynucleotide comprises a nucleic acid sequence encoding an HIF-1α polypeptide having an amino acid sequence as set forth in SEQ ID NO: 2, and said endothelial specific promoter element comprises a PPE-1-3× promoter as disclosed in PCT/IL01/01059. 
     
     
         17 . The method of  claim 1 , wherein said isolated polynucleotide comprises a nucleic acid sequence as set forth in SEQ ID NO: 1 and said endothelial specific promoter element comprises a PPE-1-3× promoter as disclosed in PCT/IL01/01059.