Gene therapy for pulmonary edema using adenovirus vectors encoding Na,K-ATPase

Abstract

Methods and compositions are provided for gene therapy for pulmonary edema in mammals by the transfer of Na,K-ATPase subunit genes to lung epithelial cells for the purpose of increasing levels of Na,K-ATPase in vivo. Recombinant adenoviral vectors mediate the transfer of the Na,K-ATPase subunit genes into lung epithelial cells. The vectors employ expression control sequences consisting of viral derived promoter elements linked to cDNAs that express Na,K-ATPase subunit genes. These genes have been shown to be capable of generating the transepithelial osmotic gradient responsible for the movement of water across epithelial membranes thereby causing pulmonary edema clearance in mammalian lungs.

1 . A method for reducing pulmonary edema of a mammalian lung that is diseased said method comprising: 
 (a) obtaining a recombinant genetic vector comprising 
 (i) an adenovirus that is deleted for nucleotide sequences encoding adenovirus proteins E1a, E1b, E3; and  
 (ii) nucleotide sequences encoding Na,K-ATPase subunits at levels that are an overexpression compared to levels in lung cells not having the genetic vector; and  
   (b) transferring the genetic vector into epithelial cells of the lung under conditions allowing overexpression of the nucleotide sequences encoding Na,K-ATPase subunits.    
     
     
         2 . The method of  claim 1 , wherein the adenovirus is high capacity and helper-virus dependent.  
     
     
         3 . The method of  claim 1 , wherein the epithelial cells are alveolar.  
     
     
         4 . The method of  claim 1  wherein the subunits encoded are a 1 , a 2 , and/or b 1 .  
     
     
         5 . A recombinant genetic vector comprising: 
 (a) an adenovirus that is deleted for nucleotide sequences encoding adenovirus proteins E1a, E1b and E3; and    (b) nucleotide sequences encoding Na,K-ATPase subunits at levels that are an overexpression compared to levels in lung cells not having the genetic vector.    
     
     
         6 . The vector of  claim 5 , further comprising regulatory elements.  
     
     
         7 . A host cell transfected with a recombinant genetic vector, said vector comprising: 
 (a) an adenovirus that is deleted for nucleotide sequences encoding adenovirus proteins E1a, E1b and E3; and    (b) nucleotide sequences encoding Na,K-ATPase subunits at levels that are an overexpression compared to levels in lung cells not having the genetic vector.