Rna interference for the treatment of heart failure

Abstract

The present invention relates to targeted RNAi for the treatment of heart failure by modulating defective cardiac Ca.sup.2+ homeostasis via decreasing expression or activity of phospholamban (PLB) using adeno-associated virus (AAV) transfection of cardiomyocytes. Methods for decreasing ventricular arrhythmias, as well as methods for overall improvement of survival from heart failure in subjects are also disclosed. Further, the present invention provides methods which can be used to diagnose susceptibility to treatment by RNAi, and includes pharmaceutical compositions, kits and vectors including an RNAi sequence.

1 . A method of treating heart failure in a subject comprising administering to a subject in need thereof a vector comprising an RNAi expression cassette, wherein the cassette comprises an RNAi sequence whose expression product decreases expression or activity of phospholamban (PLB), in an amount effective to treat heart failure of the subject. 
     
     
         2 . The method of  claim 1 , wherein the RNAi expression cassette comprises a nucleotide sequence as set forth in SEQ ID NO:1, thereby improving systolic function in the subject as compared to prior to administering the vector. 
     
     
         3 . The method of  claim 1 , wherein the vector is a virion. 
     
     
         4 . The method of  claim 1  wherein the vector is a plasmid. 
     
     
         5 . The method of  claim 3 , wherein the virion is a parvovirus. 
     
     
         6 . The method of  claim 5 , wherein the parvovirus is an adeno-associated virus (AAV). 
     
     
         7 . The method of  claim 6 , wherein the AAV is serotype 9. 
     
     
         8 . The method of  claim 1 , wherein expression of PLB mRNA is inhibited. 
     
     
         9 . The method of  claim 2 , wherein SEQ ID NO: 1 is flanked by AAV terminal repeats. 
     
     
         10 . The method of  claim 3 , wherein the genome of the virion vector is self-complementary. 
     
     
         11 - 12 . (canceled) 
     
     
         13 . A pharmaceutical composition comprising a recombinant adeno-associated virus (AAV) vector and a pharmaceutically acceptable carrier, wherein the vector comprises an RNAi expression cassette whose RNA expression product leads to decrease in expression of phospholamban (PLB) or PLB activity. 
     
     
         14 . The pharmaceutical composition of  claim 13 , wherein the RNA expression product of the RNAi expression cassette comprises a nucleotide sequence that hybridizes under stringent conditions to a nucleotide sequence of the PLB gene mRNA. 
     
     
         15 . The pharmaceutical composition of  claim 13 , wherein the RNAi expression cassette comprises a nucleotide sequence as set forth in SEQ ID NO:1. 
     
     
         16 . The pharmaceutical composition of  claim 13 , wherein the RNA expression product further comprises a target sequence as set forth in SEQ ID NO:2. 
     
     
         17 - 21 . (canceled) 
     
     
         22 . An adeno-associated virus (AAV) vector comprising at least one AAV terminal repeat, wherein the vector comprises an RNAi expression cassette, whose RNAi expression product leads to a decrease in expression of phospholamban (PLB) mRNA or PLB activity. 
     
     
         23 . The adeno-associated virus of  claim 22 , wherein the RNA expression product of the RNAi expression cassette comprises a nucleotide sequence that hybridizes under stringent conditions to a nucleotide sequence of the PLB gene mRNA, thereby transducing cardiomyocytes. 
     
     
         24 . The AAV vector of  claim 22 , wherein the RNAi expression cassette comprises a nucleotide sequence as set forth in SEQ ID NO:1. 
     
     
         25 . The AAV vector of  claim 22 , wherein the RNA expression product further comprises a target sequence as set forth in SEQ ID NO:2. 
     
     
         26 . The AAV vector of  claim 22 , wherein the AAV is serotype 9. 
     
     
         27 - 35 . (canceled) 
     
     
         36 . A kit comprising:
 i) an adeno-associated virus (AAV) vector, wherein the vector comprises a polynucleotide sequence encoding an RNAi expression product which leads to a decrease in expression of phospholamban (PLB) or PLB activity;   ii) one or more buffers;   iii) instructions on using the components of (i) and (ii); and   iv) a container comprising the components of (i), (ii) and (iii).   
     
     
         37 - 63 . (canceled)