RNA INTERFERENCE FOR THE TREATMENT OF GAIN-OF-FUNCTION DISORDERS

• US 20090118206A1
• Filed: 09/13/2004
• Published: 05/07/2009
• Est. Priority Date: 09/12/2003
• Status: Active Grant

• Current Assignee: University Of Massachusetts
• Original Assignee: University Of Massachusetts
• Inventors: Zamore, Phillip D., Aronin, Neil
• Granted Patent: US 7,947,658 B2
• Time in Patent Office: Days
• US Class Current: 514/44.R
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2510/00 Genetically modified cells

Abstract

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.

Claims

1. A method of treating a subject having or at risk for a disease characterized or caused by a gain-of-function mutant protein, comprising:

administering to said subject an effective amount of a RNAi agent targeting an allelic polymorphism within a gene encoding said mutant protein, such that sequence-specific interference of said gene occurs;

thereby treating said disease in said subject.

2. The method of claim 1, wherein said gene comprises an expanded trinucleotide repeat region.

3. The method of claim 1, wherein said mutant protein comprises an expanded polyglutamine domain.

4. The method of claim 1, wherein the disease is selected from the group consisting of Huntington'"'"'s disease, spino-cerebellar ataxia type 1, spino-cerebellar ataxia type 2, spino-cerebellar ataxia type 3, spino-cerebellar ataxia type 6, spino-cerebellar ataxia type 7, spino-cerebellar ataxia type 8, spino-cerebellar ataxia type 12, fragile X syndrome, fragile XE MR, Friedreich ataxia, myotonic dystrophy, spinal bulbar muscular disease and dentatoiubral-pallidoluysian atrophy.

5. The method of claim 4, wherein the disease is Huntington'"'"'s disease.

6. The method of claim 5, wherein the RNAi agent targets an allelic polymorphism within the gene encoding a huntingtin protein.

7. The method of claim 5, wherein the RNAi agent targets a polymorphism selected from the group consisting of P1-P5.

8. The method of claim 5, wherein the RNAi agent targets a polymorphism selected from the group consisting of P6-P43.

9. The method of claim 1, wherein the RNAi agent 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 complementary to the first strand.

10. The method of claim 1, wherein the effective amount is an amount effective to inhibit the expression or activity of the mutant protein.

11. An RNAi agent comprising a first strand comprising about 16-25 nucleotides homologous to a region of a gene encoding a gain-of-function mutant protein, said region comprising an allelic polymorphism, and a second strand comprising about 16-25 nucleotides complementary to the first strand, wherein the RNAi agent direct target-specific cleavage of a mRNA transcribed from the gene encoding the mutant protein.

12. The RNAi agent of claim 11, which targets a polymorphism within the gene encoding a Huntington protein.

13. The RNAi agent of claim 12, wherein said polymorphism is selected from the group comprising P1-P5.

14. The RNAi agent of claim 12, wherein said polymorphism is selected from the group comprising P6-P43.

15. The RNAi agent of claim 11, wherein the first strand comprises a nucleotide sequence identical to the sequence of the polymorphism.

16. The RNAi agent of claim 11, further comprising a loop portion comprising 4-11 nucleotides that connects the two strands.

17. An isolated nucleic acid molecule encoding the RNAi agent of claim 11.

18. A vector comprising the nucleic acid molecule of claim 17.

19. The vector of claim 18, which is a viral vector, retroviral vector, expression cassette, or plasmid.

20. The vector of claim 18, further comprising an RNA Polymerase III or RNA Polymerase II promoter.

21. The vector of claim 18, wherein the RNA Polymerase III promoter is the U6 or H1 promoter.

22. A host cell comprising the RNAi agent of claim 11.

23. A host cell comprising the vector of claim 18.

24. The host cell of claim 22, which is a mammalian host cell.

25. The host cell of claim 24, which is a non-human mammalian cell.

26. The host cell of claim 24, which is a human cell.

27. A composition comprising the RNAi agent of claim 11, and a pharmaceutically acceptable carrier.

28. A method for treating a disease or disorder in a subject caused by a gain-of function mutant protein, comprising identifying an allelic polymorphism within a gene encoding said mutant protein and administering to said subject an RNAi agent targeting said polymorphism such that the mutant protein is decreased, thereby treating the subject.