Products and methods for treatment of familial amyotrophic lateral sclerosis

Abstract

The present invention relates to RNA-based methods for inhibiting the expression of the superoxide dismutase 1 (SOD-1) gene. Recombinant adeno-associated viruses of the invention deliver DNAs encoding RNAs that knock down the expression of SOD-1. The methods have application in the treatment of amyotrophic lateral sclerosis.

We claim: 
     
         1 . A recombinant adeno-associated virus comprising the superoxide dismutase 1 (SOD1) shRNA-encoding DNA: 
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 1) 
                 
                     
                   GCATCATCAATTTCGAGCAGAAGGAA, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 2) 
                 
                     
                   GAAGCATTAAAGGACTGACTGAA, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 3) 
                 
                     
                   CTGACTGAAGGCCTGCATGGATT, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 4) 
                 
                     
                   CATGGATTCCATGTTCATGA, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 5) 
                 
                     
                   GCATGGATTCCATGTTCATGA, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 6) 
                 
                     
                   GGTCTGGCCTATAAAGTAGTC, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 7) 
                 
                     
                   GGGCATCATCAATTTCGAGCA, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 8) 
                 
                     
                   GCATCATCAATTTCGAGCAGA, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 9) 
                 
                     
                   GCCTGCATGGATTCCATGTTC, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 10) 
                 
                     
                   GGAGGTCTGGCCTATAAAGTA, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 11) 
                 
                     
                   GATTCCATGTTCATGAGTTTG, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 12) 
                 
                     
                   GGAGATAATACAGCAGGCTGT, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 13) 
                 
                     
                   GCTTTAAAGTACCTGTAGTGA, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 14) 
                 
                     
                   GCATTAAAGGACTGACTGAAG, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 1) 
                 
                     
                   GCATCATCAATTTCGAGCAGAAGGAA, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 2) 
                 
                     
                   GAAGCATTAAAGGACTGACTGAA, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 3) 
                 
                     
                   CTGACTGAAGGCCTGCATGGATT, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 4) 
                 
                     
                   CATGGATTCCATGTTCATGA, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 5) 
                 
                     
                   GCATGGATTCCATGTTCATGA, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 6) 
                 
                     
                   GGTCTGGCCTATAAAGTAGTC, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 7) 
                 
                     
                   GGGCATCATCAATTTCGAGCA, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 8) 
                 
                     
                   GCATCATCAATTTCGAGCAGA, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 9) 
                 
                     
                   GCCTGCATGGATTCCATGTTC, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 10) 
                 
                     
                   GGAGGTCTGGCCTATAAAGTA, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 11) 
                 
                     
                   GATTCCATGTTCATGAGTTTG, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 12) 
                 
                     
                   GGAGATAATACAGCAGGCTGT, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 13) 
                 
                     
                   GCTTTAAAGTACCTGTAGTGA, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 14) 
                 
                     
                   GCATTAAAGGACTGACTGAAG, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 15) 
                 
                     
                   TCATCAATTTCGAGCAGAA, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 16) 
                 
                     
                   TCGAGCAGAAGGAAAGTAA, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 17) 
                 
                     
                   GCCTGCATGGATTCCATGT, 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 18) 
                 
                     
                   TCACTCTCAGGAGACCATT,  
                 
                     
                   or 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 19) 
                 
                     
                   GCTTTAAAGTACCTGTAGT, 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
         wherein the recombinant adeno-associated virus genome lacks rep and cap genes. 
       
     
     
         2 . A composition comprising the recombinant adeno-associated virus of  claim 1 . 
     
     
         3 . A method of inhibiting expression of mutant SOD1 in a cell comprising contacting the cell with a recombinant adeno-associated virus of  claim 1  or the composition of  claim 2 . 
     
     
         4 . A method of delivering the SOD1 shRNA-encoding DNA GCATCATCAATTTCGAGCAGAAGGAA (SEQ ID NO:1) to a subject in need thereof, comprising administering to the subject a recombinant adeno-associated virus comprising the SOD1 shRNA-encoding DNA GCATCATCAATTTCGAGCAGAAGGAA (SEQ ID NO:1), wherein the recombinant adeno-associated virus genome lacks rep and cap genes. 
     
     
         5 . A method of delivering the SOD1 shRNA-encoding DNA GAAGCATTAAAGGACTGACTGAA (SEQ ID NO:2) to a subject in need thereof, comprising administering to the subject a recombinant adeno-associated virus comprising the SOD1 shRNA-encoding DNA GAAGCATTAAAGGACTGACTGAA (SEQ ID NO:2), wherein the recombinant adeno-associated virus genome lacks rep and cap genes. 
     
     
         6 . A method of delivering the SOD1 shRNA-encoding DNA CTGACTGAAGGCCTGCATGGATT (SEQ ID NO:3) to a subject in need thereof, comprising administering to the subject a recombinant adeno-associated virus comprising the SOD1 shRNA-encoding DNA CTGACTGAAGGCCTGCATGGATT (SEQ ID NO:3), wherein the recombinant adeno-associated virus genome lacks rep and cap genes. 
     
     
         7 . A method of delivering the SOD1 shRNA-encoding DNA CATGGATTCCATGTTCATGA (SEQ ID NO:4) to a subject in need thereof, comprising administering to the subject a recombinant adeno-associated virus comprising the SOD1 shRNA-encoding DNA CATGGATTCCATGTTCATGA (SEQ ID NO:4), wherein the recombinant adeno-associated virus genome lacks rep and cap genes. 
     
     
         8 . A method of treating amyotrophic lateral sclerosis (ALS) comprising administering a recombinant adeno-associated virus comprising the SOD1 shRNA-encoding DNA GCATCATCAATTTCGAGCAGAAGGAA (SEQ ID NO:1), wherein the recombinant adeno-associated virus genome lacks rep and cap genes. 
     
     
         9 . A method of treating ALS comprising administering a recombinant adeno-associated virus comprising the SOD1 shRNA-encoding DNA GAAGCATTAAAGGACTGACTGAA (SEQ ID NO:2), wherein the recombinant adeno-associated virus lacks rep and cap genes. 
     
     
         10 . A method of treating ALS comprising administering a recombinant adeno-associated virus comprising the SOD1 shRNA-encoding DNA CTGACTGAAGGCCTGCATGGATT (SEQ ID NO:3), wherein the recombinant adeno-associated virus lacks rep and cap genes. 
     
     
         11 . A method of treating ALS comprising administering a recombinant adeno-associated virus comprising the SOD1 shRNA-encoding DNA CATGGATTCCATGTTCATGA (SEQ ID NO:4), wherein the recombinant adeno-associated virus lacks rep and cap genes.