Adiponectin gene therapy

Abstract

Adiponectin cDNA was cloned into AAV serotypes 1, 2, and 5-based expression vectors. Virions containing these vectors were administered to the livers of rat subjects via portal vein injection. A single injection of 6×10 11 virions of the vector caused a sustained and statistically significant reduction in body weight of the treated animals compared to the control animals. This occurred in the absence of side effects. Compared to control animals, the subject rats also exhibited reduced adipose tissue mass, reduced appetite, improved insulin sensitivity, and improved glucose tolerance.

1 . A nucleic acid comprising: 
 (A) a first AAV TR;    (B) a second AAV TR; and    (C) interposed between the first and second AAV TRs, a nucleotide sequence that encodes at least a portion of an adiponectin protein that has at least one functional activity of native adiponectin.    
   
   
       2 . The nucleic acid of  claim 1 , wherein the nucleotide sequence is derived from mouse.  
   
   
       3 . The nucleic acid of  claim 1 , wherein the nucleotide sequence is derived from rat.  
   
   
       4 . The nucleic acid of  claim 1 , wherein the nucleotide sequence is derived from human.  
   
   
       5 . The nucleic acid of  claim 1 , wherein at least a portion of an adiponectin protein is an active globular domain of the adiponectin protein.  
   
   
       6 . The nucleic acid of  claim 1 , wherein the nucleotide sequence encodes a full-length adiponectin protein.  
   
   
       7 . The nucleic acid of  claim 1 , wherein at least one of the TRs is derived from AAV serotype 1.  
   
   
       8 . The nucleic acid of  claim 1 , wherein at least one of the TRs is derived from AAV serotype 2.  
   
   
       9 . The nucleic acid of  claim 1 , wherein at least one of the TRs is derived from AAV serotype 5.  
   
   
       10 . The nucleic acid of  claim 1 , wherein the nucleotide sequence further comprises an expression control sequence.  
   
   
       11 . The nucleic acid of  claim 10 , wherein the expression control sequence effects tissue-specific expression of the nucleotide sequence.  
   
   
       12 . The nucleic acid of  claim 11 , wherein the expression control sequence effects liver-specific expression of the nucleotide sequence.  
   
   
       13 . The nucleic acid of  claim 11 , wherein the expression control sequence effects muscle-specific expression of the nucleotide sequence.  
   
   
       14 . The nucleic acid of  claim 10 , wherein the expression control sequence comprises a chicken β-actin promoter operably linked to the nucleotide sequence.  
   
   
       15 . The nucleic acid of  claim 10 , wherein the expression control sequence comprises a cytomegalovirus enhancer operably linked to the nucleotide sequence.  
   
   
       16 . The nucleic acid of  claim 1 , wherein the nucleic acid is comprised within a cell.  
   
   
       17 . The nucleic acid of  claim 1 , wherein the nucleic acid is comprised within an AAV virion.  
   
   
       18 . A method of modulating adiponectin protein levels in a subject, the method comprising administering to the subject a nucleic acid comprising: 
 (A) a first AAV TR;    (B) a second AAV TR; and    (C) interposed between the first and second AAV TRs, a nucleotide sequence that encodes at least a portion of an adiponectin protein that has at least one functional activity of native adiponectin.    
   
   
       19 . The method of  claim 18 , wherein the nucleotide sequence is derived from mouse.  
   
   
       20 . The method of  claim 18 , wherein the nucleotide sequence is derived from rat.  
   
   
       21 . The method of  claim 18 , wherein the nucleotide sequence is derived from human.  
   
   
       22 . The method of  claim 18 , wherein at least a the portion of an adiponectin protein is the active globular domain of the adiponectin protein.  
   
   
       23 . The method of  claim 18 , wherein the nucleotide sequence encodes a full-length adiponectin protein.  
   
   
       24 . The method of  claim 18 , wherein the nucleotide sequence further comprises an expression control sequence.  
   
   
       25 . The method of  claim 18 , wherein the expression control sequence effects tissue-specific expression of the nucleotide sequence.  
   
   
       26 . The method of  claim 18 , wherein the expression control sequence effects liver-specific expression of the nucleotide sequence.  
   
   
       27 . The method of  claim 18 , wherein the expression control sequence effects muscle-specific expression of the nucleotide sequence.  
   
   
       28 . The method of  claim 18 , wherein the expression control sequence comprises a chicken β-actin promoter operably linked to the nucleotide sequence.  
   
   
       29 . The method of  claim 18 , wherein at least one TR is derived from AAV serotype 1.  
   
   
       30 . The method of  claim 18 , wherein at least one TR is derived from AAV serotype 2.  
   
   
       31 . The method of  claim 18 , wherein at least one TR is derived from AAV serotype 5.  
   
   
       32 . The method of  claim 18 , wherein the nucleic acid is comprised within an AAV virion.  
   
   
       33 . The method of  claim 18 , wherein the modulation of adiponectin protein  
     levels results in a reduction of weight gain in the subject.  
   
   
       34 . The method of  claim 18 , wherein modulation of adiponectin levels results in an increase in insulin sensitivity in the subject.  
   
   
       35 . The method of  claim 18 , wherein modulation of adiponectin levels results  
     in an increase in glucose tolerance in the subject.  
   
   
       36 . The method of  claim 18 , wherein modulation of adiponectin levels results  
     in a reduction of appetite in the subject.  
   
   
       37 . The method of  claim 18 , wherein the nucleic acid is administered to the subject by intravenous or intramuscular injection.