Method to induce neovascular formation and tissue regeneration

Abstract

The present invention relates, e.g., to a method for inducing arteriogenesis, lymphangiogenesis, vasculogenesis, or cardiomyogenesis, and/or for inducing mitosis or proliferation of a smooth muscle cell, a skeletal muscle cell, or a cardiomyocyte, comprising administering to a cell or tissue in need thereof a dose of a polynucleotide that encodes a vascular endothelial growth factor (VEGF), or that encodes a polypeptide comprising an active site of the VEGF. The coding sequence is operably linked to an expression control sequence; and the dose is sufficient to induce arteriogenesis, lymphangiogenesis, vasculogenesis, or cardiomyogenesis, and/or to induce mitosis or proliferation of a smooth muscle cell, a skeletal muscle cell, or a cardiomyocyte. In preferred embodiments of the invention, the method is a method of tissue regeneration, particularly of cardiomyogenesis; and the polynucleotide (or a polypeptide encoded by such a polynucleotde) is administered into the myocardium.

We claim: 
     
         1 . A method for inducing mitosis and/or proliferation of a peripheral striated muscle cell in a tissue comprising peripheral striated muscle cells, in a subject in need thereof, comprising administering directly to the cell or tissue a dose of a polynucleotide that encodes the vascular endothelial growth factor, VEGF 1-165, wherein the coding sequence is operably linked to a CMV promoter and is in a plasmid vector, the dose being in a sufficient amount to induce mitosis and/or proliferation of a peripheral striated muscle cell in a tissue comprising striated muscle cells, wherein the dose is equivalent to the at least about 0.04 mg/kg that, when administered to a subject in need of the induction of cardiomyogenesis, induces cardiomyogenesis. 
     
     
         2 . The method of  claim 1 , wherein the VEGF 1-165 has the amino acid sequence: 
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 1) 
                 
                     
                   Ala Pro Met Ala Glu Gly Gly Gly Gln Asn 
                 
                     
                     
                 
                     
                   His His Glu Val Val Lys Phe Met Asp Val 
                 
                     
                     
                 
                     
                   Tyr Gln Arg Ser Tyr Cys His Pro Ile Glu 
                 
                     
                     
                 
                     
                   Thr Leu Val Asp Ile Phe Gln Glu Tyr Pro 
                 
                     
                     
                 
                     
                   Asp Glu Ile Glu Tyr Ile Phe Lys Pro Ser 
                 
                     
                     
                 
                     
                   Cys Val Pro Leu Met Arg Cys Gly Gly Cys 
                 
                     
                     
                 
                     
                   Cys Asn Asp Glu Gly Leu Glu Cys Val Pro 
                 
                     
                     
                 
                     
                   Thr Glu Glu Ser Asn Ile Thr Met Gln Ile 
                 
                     
                     
                 
                     
                   Met Arg Ile Lys Pro His Gln Gly Gln His 
                 
                     
                     
                 
                     
                   Ile Gly Glu Met Ser Phe Leu Gln His Asn 
                 
                     
                     
                 
                     
                   Lys Cys Glu Cys Arg Pro Lys Lys Asp Arg 
                 
                     
                     
                 
                     
                   Ala Arg Gln Glu Asn Pro Cys Gly Pro Cys 
                 
                     
                     
                 
                     
                   Ser Glu Arg Arg Lys His Leu Phe Val Gln 
                 
                     
                     
                 
                     
                   Asp Pro Gln Thr Cys Lys Cys Ser Cys Lys 
                 
                     
                     
                 
                     
                   Asn Thr Asp Ser Arg Cys Lys Ala Arg Gln 
                 
                     
                     
                 
                     
                   Leu Glu Leu Asn Glu Arg Thr Cys Arg Cys 
                 
                     
                     
                 
                     
                   Asp Lys Pro Arg Arg. 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         3 . The method of  claim 1 , wherein the mitosis and/or proliferation is localized. 
     
     
         4 . The method of  claim 1 , wherein the tissue comprising striated muscle cells is induced in normoperfused tissue. 
     
     
         5 . The method of  claim 1 , wherein the tissue comprising striated muscle cells is induced in ischemic tissue. 
     
     
         6 . The method of  claim 1 , wherein the cell or tissue is eukaryotic. 
     
     
         7 . The method of  claim 1 , wherein the cell or tissue is mammalian. 
     
     
         8 . The method of  claim 1 , wherein the cell or tissue is pig, rabbit, sheep or human. 
     
     
         9 . The method of  claim 1 , wherein the cell or tissue is human. 
     
     
         10 . The method of  claim 1 , wherein the polynucleotide is a genomic DNA, a cDNA, or a messenger RNA. 
     
     
         11 . The method of  claim 10 , wherein the polynucleotide encodes the polypeptide represented by SEQ ID NO: 1. 
     
     
         12 . The method of  claim 11 , wherein the polynucleotide is a cDNA. 
     
     
         13 . The method of  claim 1 , wherein the polynucleotide is administered to the cell or tissue in a liposome. 
     
     
         14 . The method of  claim 1 , wherein the subject exhibits complications of diabetes. 
     
     
         15 . The method of  claim 14 , wherein the complications are peripheral artery disease (PAD), diabetic neuropathy, vasa-vasorum disease and/or severe limb ischemia. 
     
     
         16 . The method of  claim 1 , wherein the subject has PAD and/or limb ischemia. 
     
     
         17 . The method of  claim 16 , wherein the limb ischemia is severe limb ischemia. 
     
     
         18 . The method of  claim 1 , wherein the subject is human. 
     
     
         19 . The method of  claim 1 , wherein the polynucleotide is in the form of a pharmaceutical composition. 
     
     
         20 . The method of  claim 1 , wherein the dose of polynucleotide is administered multiple times.