Antisense oligonucleotide directed removal of proteolytic cleavage sites, the hchwa-d mutation, and trinucleotide repeat expansions

Abstract

Described are methods for removing a proteolytic cleavage site, the HCHWA-D mutation or the amino acids encoded by a trinucleotide repeat expansion from a protein comprising providing a cell that expresses pre-mRNA encoding the protein with an anti-sense oligonucleotide that induces skipping of the exonic sequence that comprises the proteolytic cleavage site, HCHWA-D mutation or trinucleotide repeat expansion, respectively, the method further comprising allowing translation of mRNA produced from the pre-mRNA.

What is claimed is: 
     
         1 . A method for treating an individual suffering from a disease that is associated with a mutant gene comprising a trinucleotide repeat expansion when compared to the gene of a normal individual, the method comprising:
 administering to the individual a therapeutically effective amount of one or more anti-sense oligonucleotides that induce skipping of an exonic sequence that comprises the trinucleotide repeat expansion.   
     
     
         2 . The method according to  claim 1 , wherein the mutant gene is the causative gene of a polyglutamine disorder. 
     
     
         3 . The method according to  claim 2 , wherein the gene is the ATXN3 gene and exonic sequences from exons 9 and 10 thereof are skipped. 
     
     
         4 . A method for removing amino acids encoded by a trinucleotide repeat expansion from a mutant protein, the method comprising:
 providing a cell that expresses pre-mRNA encoding the mutant protein with an anti-sense oligonucleotide that induces skipping of an exonic sequence that comprises the trinucleotide repeat expansion, and   allowing translation of mRNA produced from the pre-mRNA.   
     
     
         5 . The method according to  claim 4 , wherein the trinucleotide repeat expansion is a polyglutamine expansion. 
     
     
         6 . At least one oligonucleotide of between 14-40 nucleotides that induces skipping of an exonic sequence that comprises a trinucleotide repeat expansion in a pre-mRNA. 
     
     
         7 . The at least one oligonucleotide of  claim 6 , wherein the at least one oligonucleotide binds to the pre-mRNA of protein to form a double-stranded nucleic acid complex and wherein the at least one oligonucleotide is chemically modified to render the double-stranded nucleic acid complex RNAse H resistant. 
     
     
         8 . The at least one oligonucleotide of  claim 6 , comprising:
 a first oligonucleotide that induces skipping of an exonic sequence from exon 9 of ATXN3, and   a second oligonucleotide that induces skipping of an exonic sequence from exon 10 of ATXN3 comprising a trinucleotide repeat expansion.   
     
     
         9 . A method for treating an individual afflicted with HCHWA-D mutation (hereditary cerebral hemorrhage with amyloidosis, Dutch type), the method comprising:
 administering to the individual:
 a therapeutically effective amount of one or more anti-sense oligonucleotides that induce skipping of the exonic sequence that comprises the HCHWA-D mutation, and/or 
 a therapeutically effective amount of one or more cells comprising the oligonucleotides. 
   
     
     
         10 . The method according to  claim 9 , wherein the oligonucleotide induces skipping of an exonic sequence corresponding to exon 16 of APP751. 
     
     
         11 . A method for removing the HCHWA-D mutation from mutant APP protein, the method comprising:
 providing a cell that expresses pre-mRNA encoding the mutant APP protein with an anti-sense oligonucleotide that induces skipping of the exonic sequence that comprises the HCHWA-D mutation, and   allowing translation of mRNA produced from the pre-mRNA in the cell.   
     
     
         12 . The method according to  claim 11 , wherein the oligonucleotide induces skipping of an exonic sequence corresponding to exon 16 of APP751. 
     
     
         13 . At least one oligonucleotide of between 14-40 nucleotides that induces skipping of an exonic sequence that comprises the HCHWA-D mutation of mutant APP protein. 
     
     
         14 . The at least one oligonucleotide of  claim 13 ,
 wherein the oligonucleotide binds to the pre-mRNA of the mutant APP protein to form a double-stranded nucleic acid complex, and   wherein the oligonucleotide is chemically modified to render the double-stranded nucleic acid complex RNAse H resistant.   
     
     
         15 . The at least one oligonucleotide of  claim 13 , wherein the at least one oligonucleotide comprises an oligonucleotide that induces skipping of an exonic sequence corresponding to exon 16 of APP751.