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-1alpha, the polypeptide being stably expressed and constitutively active. Isolated polypeptides encoded by same are also disclosed and uses thereof.

1 - 21 . (canceled) 
     
     
         22 . An isolated polynucleotide comprising a nucleic acid sequence encoding a polypeptide having an amino acid sequence of HIF-1α, said polypeptide being stably expressed and constitutively active in endothelial cells. 
     
     
         23 . The isolated polynucleotide of  claim 22 , wherein said amino acid sequence is as set forth in SEQ ID NO: 2. 
     
     
         24 . The isolated polynucleotide of  claim 22 , wherein said nucleic acid sequence is as set forth in SEQ ID NO:1. 
     
     
         25 . An isolated polynucleotide comprising a nucleic acid sequence encoding the amino acid sequence as set forth in SEQ ID NO: 2 
     
     
         26 . An isolated polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 2. 
     
     
         27 . A nucleic acid construct comprising the isolated polynucleotide of  claim 25 . 
     
     
         28 . The nucleic acid construct of  claim 27 , further comprising a cis-regulatory element. 
     
     
         29 . The nucleic acid construct of  claim 28 , wherein said cis-regulatory element comprises a promoter element. 
     
     
         30 . The nucleic acid construct of  claim 29 , wherein said promoter element is an endothelial specific promoter element. 
     
     
         31 . The nucleic acid construct of  claim 30 , wherein said endothelial specific promoter element comprises a PPE-1 promoter. 
     
     
         32 . The nucleic acid construct of  claim 30 , wherein said endothelial specific promoter is a PPE-1 3X promoter. 
     
     
         33 . The nucleic acid construct of  claim 27 , further comprising a hypoxia response element as set forth in SEQ ID NO: 5. 
     
     
         34 . The nucleic acid construct of  claim 32 , further comprising a hypoxia response element as set forth in SEQ ID NO: 5. 
     
     
         35 . A cell comprising the nucleic acid construct of  claim 27 . 
     
     
         36 . A pharmaceutical composition comprising as an active agent the nucleic acid construct of  claim 27  and a pharmaceutically acceptable carrier. 
     
     
         37 . The pharmaceutical composition of  claim 36 , wherein said nucleic acid construct, further comprising a cis-regulatory element. 
     
     
         38 . The pharmaceutical composition of  claim 37 , wherein said cis-regulatory element comprises a promoter element. 
     
     
         39 . The pharmaceutical composition of  claim 38 , wherein said promoter element is an endothelial specific promoter element. 
     
     
         40 . The pharmaceutical composition of  claim 39 , wherein said endothelial specific promoter element comprises a PPE-1 promoter. 
     
     
         41 . The pharmaceutical composition of  claim 39 , wherein said endothelial specific promoter is a PPE-1 3X promoter. 
     
     
         42 . The pharmaceutical composition of  claim 36 , further comprising a hypoxia response element as set forth in SEQ ID NO: 5. 
     
     
         43 . The pharmaceutical composition of  claim 41 , further comprising a hypoxia response element as set forth in SEQ ID NO: 5. 
     
     
         44 . A pharmaceutical composition comprising as an active agent the polypeptide of  claim 26  and a pharmaceutically acceptable carrier. 
     
     
         45 . 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 the polypeptide of  claim 26 , thereby treating said medical condition in said subject. 
     
     
         46 . The method of  claim 45 , 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. 
     
     
         47 . A method of treating a medical condition associated with hypoxia or ischemia, reduced tissue perfusion inclusion or ‘low flow’, the method comprising expressing in a subject in need thereof a therapeutically effective amount of a nucleic acid construct of  claim 27 , thereby treating said medical condition in said subject. 
     
     
         48 . The method of  claim 47 , wherein said nucleic acid construct, further comprising a cis-regulatory element. 
     
     
         49 . The method of  claim 48 , wherein said cis-regulatory element comprises a promoter element. 
     
     
         50 . The method of  claim 49 , wherein said promoter element is an endothelial specific promoter element. 
     
     
         51 . The method of  claim 50 , wherein said endothelial specific promoter element comprises a PPE-1 promoter. 
     
     
         52 . The method of  claim 50 , wherein said endothelial specific promoter is a PPE-1 3X promoter. 
     
     
         53 . The pharmaceutical composition of  claim 47 , further comprising a hypoxia response element as set forth in SEQ ID NO: 5. 
     
     
         54 . The pharmaceutical composition of  claim 52 , further comprising a hypoxia response element as set forth in SEQ ID NO: 5. 
     
     
         55 . The method of  claim 47 , 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. 
     
     
         56 . A method of treating a medical condition associated with hypoxia or ischemia, reduced tissue perfusion inclusion or ‘low flow’, the method comprising expressing in a subject in need thereof a therapeutically effective amount of a nucleic acid construct of  claim 32 , thereby treating said medical condition in said subject. 
     
     
         57 . The method of  claim 56 , wherein said nucleic acid construct further comprises a hypoxia response element as set forth in SEQ ID NO: 5. 
     
     
         58 . The method of  claim 56 , 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.