RNA interference for the treatment of gain-of-function disorders
The present invention relates to the discovery of an effective treatment for a variety of gain-of-function diseases, in particular, Huntington's disease (HD). The present invention utilizes RNA Interference technology (RNAi) against polymorphic regions in the genes encoding various gain-of-function mutant proteins resulting in an effective treatment for the gain-of-function disease.
1. A method of treating a subject having or at risk for Huntington's disease characterized or caused by a mutant huntingtin (htt) protein, comprising:
administering to the subject an effective amount of an siRNA or shRNA targeting a heterozygous single nucleotide polymorphism (SNP) within a gene encoding the mutant htt protein, wherein the SNP has an allelic frequency of at least 30% in a sample population and is present at a genomic site selected from the group consisting of RS362331, RS4690077, RS362268, RS362267, RS362307, and RS362306, such that sequence-specific interference of said gene occurs;
thereby treating said disease in said subject.
2. A method of silencing a target mRNA encoding a mutant huntingtin (htt) protein in a cell, comprising contacting the cell with an effective amount of an siRNA or shRNA targeting a heterozygous single nucleotide polymorphism (SNP) within the target mRNA, such that RNA silencing of said mRNA occurs, wherein the SNP has an allelic frequency of at least 30% in a sample population and is present at a genomic site selected from the group consisting of RS362331, RS4690077, RS362268, RS362267, RS362307, and RS362306.
3. The method of claim 1 or 2 , wherein the SNP is present at genomic site RS362307.
4. The method of claim 1 or 2 , wherein the siRNA comprises a first strand comprising about 16-25 nucleotides homologous to a region of the gene comprising the polymorphism and a second strand comprising about 16-25 nucleotides substantially complementary to the first strand such that the first and second strands are capable of annealing together.
5. The method of claim 1 or 2 , wherein the SNP is present at genomic site RS362331.
6. The method of claim 1 or 2 , wherein the gene or target mRNA encoding the mutant htt protein comprises a cytosine at genomic site RS362331.
7. The method of claim 1 or 2 , wherein the gene or target mRNA encoding the mutant htt protein comprises a uracil at genomic site RS362331.
8. The method of claim 1 or 2 , wherein the antisense strand of said siRNA or shRNA is complementary to the SNP and wherein said siRNA or shRNA is capable of substantially silencing the mutant huntingtin protein without substantially silencing the corresponding wild-type huntingtin protein.
9. The method of claim 1 or 2 , wherein the sample population is of Western European origin.
10. The method of claim 1 or 2 , wherein the siRNA comprises a lipophilic moiety.
11. The method of claim 10 , wherein the lipophilic moiety is a cholesterol moiety.
12. A method of treating a subject having or at risk for Huntington's disease characterized or caused by a mutant huntingtin (htt) protein, comprising administering to the subject an effective amount of a siRNA targeting a heterozygous single nucleotide polymorphism (SNP) within a gene encoding the mutant htt protein, such that RNA silencing of said gene occurs, wherein the siRNA comprises (i) an antisense strand or a variant thereof having sufficient complementarity to the SNP at genomic site RS362307 to direct target-specific RNA interference of the gene encoding the mutant htt protein; and (ii) a sense strand or a variant thereof that is substantially complementary to the antisense strand, such that the sense and antisense strands are capable of annealing together, said variant comprising at least one nucleotide analog or backbone modification.
13. The method of claim 12 , wherein the siRNA comprises a lipophilic moiety.
14. The method of claim 13 , wherein the lipophilic moiety is a cholesterol moiety.
15. A method of treating a subject having or at risk for Huntington's disease characterized or caused by a mutant huntingtin (htt) protein, comprising administering to the subject an effective amount of a siRNA targeting a heterozygous single nucleotide polymorphism (SNP) within a gene encoding the mutant htt protein, such that RNA silencing of said gene occurs, wherein the siRNA comprises (i) an antisense strand or a variant thereof having sufficient complementarity to the SNP at genomic site RS362331 or RS363125 to direct target-specific RNA interference of the gene encoding the mutant htt protein; and (ii) a sense strand or a variant thereof that is substantially complementary to the antisense strand, such that the sense and antisense strands are capable of annealing together, said variant comprising at least one nucleotide analog or backbone modification.
16. A method of treating a subject having or at risk for a disease characterized or caused by a mutant huntingtin (htt) protein, comprising:
administering to the subject an effective amount of an siRNA or shRNA targeting a heterozygous single nucleotide polymorphism located at genomic site RS362307 within the gene encoding the mutant htt protein, such that sequence-specific interference of said gene occurs;
thereby treating said disease in said subject.
17. A method of silencing a target mRNA encoding a mutant huntingtin (htt) protein in a cell, comprising contacting the cell with an effective amount of an siRNA or shRNA targeting a heterozygous single nucleotide polymorphism located at genomic site RS362307 within the target mRNA, such that RNA silencing of said mRNA occurs.