Single nucleotide polymorphism (SNP) targeting therapies for the treatment of huntington'S disease

• US 8,987,222 B2
• Filed: 04/08/2010
• Issued: 03/24/2015
• Est. Priority Date: 04/08/2009
• Status: Active Grant

• Current Assignee: University Of Massachusetts
• Original Assignee: University Of Massachusetts
• Inventors: Aronin, Neil, Pfister, Edith, Zamore, Phillip D.
• Primary Examiner(s): Bowman, Amy
• Application Number:
  US13/263,961
• Publication Number:
  US 20120136039A1
• Time in Patent Office: 1,811 Days
• Field of Search: None
• US Class Current: 514/44
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• 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 2310/11 Antisense
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2320/34 Allele or polymorphism specific uses

Abstract

The present invention relates to the discovery of (SNPs) significantly associated with Huntington'"'"'s disease (HD). The present invention utilizes RNA silencing technology (e.g. RNAi) against such SNPs optimally combined with select additional SNP targeting silencing agents, thereby resulting in an effective treatment of significantly-sized patient populations. Silencing agents having enhanced discriminatory properties are also featured.

Claims

1. A method of silencing mutant huntingtin (htt) mRNA in a HD patient population, comprising administering to said patient population an effective amount of a first RNA silencing agent targeting a HD-associated htt single nucleotide polymorphism (SNP) in combination with one or more RNA silencing agents targeting one or more other htt SNPs, such that RNA silencing of said mRNA occurs, wherein the HD-associated htt SNP is the U isoform of rs362307, and wherein at least one other htt SNP has a frequency of heterozygosity of at least 20% or more in a sample population.

2. The method of claim 1, wherein the other htt SNPs are as set forth in Table 2.

3. The method of claim 1, wherein the htt SNP having a frequency of heterozygosity of at least 20% or more in a sample population is selected from the group consisting of rs4690074, rs362336, rs362331, rs362273, rs362272, rs362306, rs362268, and rs362267.

4. The method of claim 2, wherein the patient population is of US or Western European origin.

6. The method of claim 1 or claim 5, further comprising identifying the sequence of the nucleotide located at one or more htt SNPs in the mutant htt mRNA of the HD patients in the HD patient population.

9. The method of claim 1 or claim 5, wherein the RNA silencing agent is selected from the group consisting of a siRNA, a shRNA, or a vector encoding a shRNA.

10. The method of claim 1 or claim 5, wherein a nucleotide complementary to the SNP in the mutant htt mRNA is located at position 10 relative to the 5′

end of the antisense strand of the RNA silencing agent.

11. The method of claim 10, wherein the RNA silencing agent further comprises a mismatch with respect to both the mutant htt mRNA and the wild-type htt mRNA at one or more positions located within the seed sequence of the RNA silencing agent.

12. The method of claim 11, wherein said one or more positions are selected from the group consisting of position 2, position 3, position 4, position 5, position 6, and position 7 relative to the 5′

end of the antisense strand of the RNA silencing agent.

13. The method of claim 12, wherein said position is position 5.

14. The method of claim 12, wherein said position is position 6.

17. The method of claim 1, wherein the other htt SNP is RS362273.

19. The method of claim 17, further comprising administering to said patient population an effective amount of an RNA silencing agent targeting rs363125.

21. The method of claim 1 or claim 5, wherein the RNA silencing agent targeting the U isoform of rs362307 contains a guide strand comprising SEQ ID NO:

219 or SEQ ID NO;

220.

5. A method of silencing mutant htt mRNA in 70% or more HD patients in a HD patient population, comprising administering to said HD patient population a plurality of RNA silencing agents, wherein a first RNA silencing agent targets the U isoform of HD-associated SNP RS362307, a second RNA silencing agent targets a htt single nucleotide polymorphism (SNP) having a frequency of heterozygosity of at least 20% or more in a sample HD patient population, and a third RNA silencing agent targets a third htt SNP, such that RNA silencing of said mRNA occurs in 70% or more patients in the HD patient population.

7. The method of claim 5, wherein the RNA silencing agents target SNP1, SNP2, and SNP3 as set forth in Table 2.

8. The method of claim 5, wherein the second RNA silencing agent targets rs362273.

15. The method of claim 5, wherein the guide strand of the first RNA silencing agent comprises SEQ ID NO:

220, wherein the guide strand of the second RNA silencing agent comprises SEQ ID NO;

210 or SEQ ID NO;

211, and wherein the guide strand of the third RNA silencing agent comprises SEQ ID NO;

230 or SEQ ID NO;

233.

16. The method of claim 5, wherein the guide strand of the first RNA silencing agent comprises SEQ ID NO:

219, wherein the guide strand of the second RNA silencing agent comprises SEQ ID NO;

210 or SEQ ID NO;

211, and wherein the guide strand of the third RNA silencing agent comprises SEQ ID NO;

230 or SEQ ID NO;

233.

18. The method of claim 5, wherein the second RNA silencing agent targets a htt SNP selected from the group consisting of rs4690074, rs362336, rs362331, rs362273, rs362272, rs362306, rs362268, and rs362267.

20. The method of claim 8, wherein the third RNA silencing agent targets rs363125.