Promoters exhibiting endothelial cell specificity and methods of using same for regulation of angiogenesis

Abstract

Isolated polynucleotide sequences exhibiting endothelial cell specific promoter activity, novel cis regulatory elements and methods of use thereof enabling treatment of diseases characterized by aberrant neovascularization or cell growth are disclosed.

1 . A method of inhibiting angiogenesis in an endothelial tissue of a subject in need thereof, the method comprising administering to said subject 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,   wherein said subject is being treated with radiation therapy.   
     
     
         2 . The method of  claim 1 , wherein said radiation therapy is administered concomitantly with said nucleic acid construct. 
     
     
         3 . The method of  claim 1 , wherein said nucleic acid construct is administered prior to treatment with said radiation therapy. 
     
     
         4 . The method of  claim 1 , wherein said apoptosis inducing factor is a pro-apoptotic gene. 
     
     
         5 . The method of  claim 1 , wherein said enhancer element further includes at least one copy of the sequence set forth in SEQ ID NO:6. 
     
     
         6 . The method 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. 
     
     
         7 . The method of  claim 1 , wherein said enhancer element is as set forth in SEQ ID NO: 7. 
     
     
         8 . The method of  claim 1 , wherein said promoter is an endothelial specific promoter element. 
     
     
         9 . The method of  claim 8 , wherein said endothelial specific promoter element comprises at least one copy of the PPE-1 promoter. 
     
     
         10 . The nucleic acid construct of  claim 9 , wherein said PPE-1 promoter element comprises the nucleic acid sequence as set forth in SEQ ID NO: 1. 
     
     
         11 . The nucleic acid construct of  claim 8 , wherein said endothelial specific promoter element comprises at least one copy of the PPE-1-3×promoter. 
     
     
         12 . The method of  claim 1 , wherein said nucleic acid construct further comprises a hypoxia response element. 
     
     
         13 . The method of  claim 12 , wherein said hypoxia response element includes at least one copy of the sequence set forth in SEQ ID NO: 5. 
     
     
         14 . The method of  claim 1 , further comprising an adenovirus vector. 
     
     
         15 . The nucleic acid construct of  claim 14 , wherein said adenovirus vector is an adenovirus serotype 5 vector. 
     
     
         16 . The method of  claim 15 , wherein said tissue is a tumor mass. 
     
     
         17 . A method of inhibiting angiogenesis in an endothelial tissue of a subject in need thereof, the method comprising administering to said subject 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 and wherein said subject is being treated with radiation therapy.   
     
     
         18 . The method of  claim 17 , wherein said radiation therapy is administered concomitantly with said nucleic acid construct. 
     
     
         19 . The method of  claim 17 , wherein said nucleic acid construct is administered prior to treatment with said radiation therapy. 
     
     
         20 . The method of  claim 17 , wherein said apoptosis inducing element is a pro-apoptotic gene. 
     
     
         21 . The method of  claim 17 , wherein said tissue is a tumor mass.