Promoters exhibiting endothelial cell specificity and methods of using same

Abstract

An isolated polynucleotide functional as a promoter in eukaryotic cells is disclosed. The isolated polynucleotide includes an endothelial specific enhancer element as detailed herein. Further disclosed is a method of expressing a nucleic acid sequence of interest in endothelial cells.

1 . A nucleic acid construct comprising:
 (a) an isolated polynucleotide comprising an enhancer element including at least one copy of the sequence set forth in SEQ ID NO:8;   (b) a promoter functional in eukaryotic cells; and   (c) a nucleic acid sequence encoding an apoptosis inducing factor under the control of said promoter.   
     
     
         2 . The nucleic acid construct of  claim 1 , wherein said apoptosis inducing factor is a pro-apoptotic gene. 
     
     
         3 . The nucleic acid construct of  claim 1 , wherein said enhancer element further includes at least one copy of the sequence set forth in SEQ ID NO:6. 
     
     
         4 . The nucleic acid construct of  claim 1 , wherein said enhancer element includes one copy of the sequence set forth in SEQ ID NO:8 and at least two copies of the sequence set forth in SEQ ID NO:6. 
     
     
         5 . The nucleic acid construct of  claim 1 , wherein said enhancer element is as set forth in SEQ ID NO: 7. 
     
     
         6 . The nucleic acid construct of  claim 1 , wherein said promoter is an endothelial specific promoter element. 
     
     
         7 . The nucleic acid construct of  claim 6 , wherein said endothelial specific promoter element comprises at least one copy of the PPE-1 promoter. 
     
     
         8 . The nucleic acid construct of  claim 7 , wherein said PPE-1 promoter element comprises the nucleic acid sequence as set forth in SEQ ID NO: 1. 
     
     
         9 . The nucleic acid construct of  claim 6 , wherein said endothelial specific promoter element comprises at least one copy of the PPE-1-3× promoter. 
     
     
         10 . The nucleic acid construct of  claim 1 , further comprising a hypoxia response element. 
     
     
         11 . The nucleic acid construct of  claim 10 , wherein said hypoxia response element includes at least one copy of the sequence set forth in SEQ ID NO: 5. 
     
     
         12 . The nucleic acid construct of  claim 1 , further comprising an adenovirus vector. 
     
     
         13 . The nucleic acid construct of  claim 12 , wherein said adenovirus vector is an adenovirus serotype 5 vector. 
     
     
         14 . A mammalian cell transformed with the nucleic acid construct of  claim 1 . 
     
     
         15 . A method of inhibiting angiogenesis in an endothelial tissue of a subject in need thereof, the method comprising administering to said subject the nucleic acid construct of  claim 1 . 
     
     
         16 . The method of  claim 15 , wherein said tissue is a tumor mass. 
     
     
         17 . A recombinant adenovirus vector comprising:
 (a) an isolated polynucleotide comprising an enhancer element including at least one copy of the sequence set forth in SEQ ID NO:7;   (b) a PPE-1 promoter functional in eukaryotic cells as set forth in SEQ ID NO: 1;   (c) a nucleic acid sequence encoding an apoptosis inducing factor under the control of said promoter; and   (d) a hypoxia response element as set forth in SEQ ID NO: 5,   wherein said adenovirus vector is an Ad serotype 5 vector.   
     
     
         18 . The adenovirus vector of  claim 17 , wherein said apoptosis inducing element is a pro-apoptotic gene. 
     
     
         19 . A method of inhibiting angiogenesis in an endothelial tissue of a subject in need thereof, the method comprising administering to said subject the recombinant adenovirus vector of  claim 17 . 
     
     
         20 . The method of  claim 18 , wherein said tissue is a tumor mass.