Vectors and methods for regenerative therapy

Abstract

An expression vector capable of disrupting the silencing of cell cycle genes in adult cells, such as adult cardiac myocytes and other quiescent cells in terminally differentiated tissues, comprising: (a) a nucleic acid sequence encoding lysine-specific demethylase 4D (KDM4D); (b) a promoter that induces or effects overexpression of KDM4D, wherein the promoter is operably linked to the nucleic acid sequence; and (c) a regulatory element that inducibly represses the overexpression of KDM4D. The vector can be administered to a subject in a method for inducing tissue-specific hyperplasia in a mammal, including cardiomyocyte proliferation. The method provides for regenerative therapy, including improving cardiac function after myocardial infarct and other forms of cardiac damage.

1 .- 20 . (canceled) 
     
     
         21 . A method for inducing cardiomyocyte (CM) hyperplasia comprising delivering a transgene encoding exogenous lysine-specific demethylase 4D (KDM4D) to CMs. 
     
     
         22 . The method of  claim 21 , wherein the KDM4D comprises an amino acid sequence of SEQ ID NO:1. 
     
     
         23 . The method of  claim 21 , wherein the method comprises contacting the CMs with an expression vector comprising:
 (a) a nucleic acid sequence encoding KDM4D;   (b) a cardiac-specific promoter capable of effecting overexpression of KDM4D only in cardiac tissue, wherein the promoter is operably linked to the nucleic acid sequence; and   (c) a regulatory element that inducibly represses the overexpression of KDM4D.   
     
     
         24 . The method of  claim 23 , wherein the expression vector is a viral vector that infects quiescent cells. 
     
     
         25 . The method of  claim 24 , wherein the viral vector is an adeno-associated virus (AAV) vector. 
     
     
         26 . The method of  claim 25 , wherein the AAV vector is AAV6 or AAV9. 
     
     
         27 . The method of  claim 21 , wherein the CMs are adult CMs (ACMs). 
     
     
         28 . A method of improving cardiac function in a mammal comprising delivering a transgene encoding exogenous KDM4D to the mammal. 
     
     
         29 . A method of proliferating CMs comprising culturing CMs with a transgene encoding exogenous KDM4D under conditions effective to induce CM hyperplasia. 
     
     
         30 . A method of promoting cardiac regeneration in a subject in need thereof comprising reducing lysine 9 of histone H3 (H3K9me3) levels in CMs. 
     
     
         31 . The method of  claim 30 , wherein the reducing comprises contacting the CMs with an expression vector comprising:
 (a) a nucleic acid sequence encoding KDM4D;   (b) a cardiac-specific promoter capable of effecting overexpression of KDM4D only in cardiac tissue, wherein the promoter is operably linked to the nucleic acid sequence; and   (c) a regulatory element that inducibly represses the overexpression of KDM4D.   
     
     
         32 . The method of  claim 31 , wherein the KDM4D comprises an amino acid sequence of SEQ ID NO:1. 
     
     
         33 . The method of  claim 31 , wherein the expression vector is a viral vector that infects quiescent cells. 
     
     
         34 . The method of  claim 33 , wherein the viral vector is an adeno-associated virus (AAV) vector. 
     
     
         35 . The method of  claim 34 , wherein the AAV vector is AAV6 or AAV9. 
     
     
         36 . The method of  claim 31 , wherein the expression vector is delivered by delivering CMs that contain the expression vector. 
     
     
         37 . The method of  claim 30 , wherein the reducing comprises delivering a transgene encoding exogenous KDM4D. 
     
     
         38 . The method of  claim 37 , wherein delivery is systemic. 
     
     
         39 . The method of  claim 37 , wherein delivery is intravenous. 
     
     
         40 . The method of  claim 37 , wherein delivery is by intra-myocardial injection.