GNAQ targeted dsRNA compositions and methods for inhibiting expression

1. A double-stranded ribonucleic acid (dsRNA) for inhibiting expression of a G-alpha q subunit (GNAQ) of a heterotrimeric G gene, comprising a sense strand and an antisense strand complementary to at least the first 11 nucleotides of SEQ ID NO:1421, wherein each strand is at least 15 nucleotides in length.

2. The dsRNA of claim 1 , wherein the sense strand consists of SEQ ID NO:1579 (cAAuuuGcAuAAuAcuAAudTdT) and the antisense strand consists of SEQ ID NO:1580 (AUuAGuAUuAUGcAAAUUGdTdT) wherein each strand includes 2′-O-methyl ribonucleotides as indicated by a lower case letter “c” and “u”.

3. The dsRNA of claim 1 , wherein the antisense strand is complementary to at least 15 contiguous nucleotides of SEQ ID NO:1421.

4. The dsRNA of claim 1 , wherein the sense strand comprises 15 or more contiguous nucleotides of SEQ ID NO:1421 or SEQ ID NO:1579.

5. The dsRNA of claim 1 , wherein the antisense strand comprises 15 or more contiguous nucleotides of SEQ ID NO:1422 or SEQ ID NO:1580.

6. The dsRNA of claim 1 , wherein the sense strand sequence comprises SEQ ID NO:1421 and the antisense strand sequence comprises of SEQ ID NO:1422.

7. The dsRNA of claim 1 , wherein administration of 0.1 nM of the dsRNA to a A375 cell results in about 66% inhibition of GNAQ mRNA expression as measured by a real time PCR assay or administration of 1 nM of the dsRNA to a A375 cell results in about 61% inhibition of GNAQ mRNA expression as measured by a real time PCR assay or administration of 1 nM of the dsRNA to a A579 cell results in about 82% inhibition of GNAQ mRNA expression as measured by a real time PCR assay or administration of 10 nM of the dsRNA to a OMM1.3 cell results in about 42% inhibition of GNAQ mRNA expression as measured by a real time PCR assay or administration of the dsRNA to a UMEL202 cell results in about 81% inhibition of GNAQ mRNA expression as measured by a real time PCR assay.

8. The dsRNA of claim 1 , wherein the antisense strand is complementary to 16, 17, or 18 nucleotides of SEQ ID NO:1421.

9. The dsRNA of claim 1 , wherein the antisense strand is complementary to 19 nucleotides of SEQ ID NO:1421.

10. The dsRNA of claim 1 , wherein each strand is no more than 30 nucleotides in length.

11. The dsRNA of claim 1 , wherein at least one strand comprises a 3′ overhang of at least 1 nucleotide.

12. The dsRNA of claim 1 , wherein each strand comprises a 3′ overhang consisting of 2 nucleotides.

13. The dsRNA of claim 1 , wherein each strand comprises a 3′ overhang consisting of dTdT.

14. The dsRNA of claim 1 or claim 6 , comprising a modification that causes the dsRNA to have increased stability in a biological sample.

15. The dsRNA of claim 1 or claim 6 , wherein said dsRNA comprises at least one modified nucleotide.

16. The dsRNA of claim 14 , wherein said modified nucleotide is selected from the group of: a 2′-O-methyl modified nucleotide, a nucleotide comprising a 5′-phosphorothioate group, and a terminal nucleotide linked to a cholesteryl derivative or dodecanoic acid bisdecylamide group.

17. The dsRNA of claim 14 , wherein said modified nucleotide is chosen from the group of: a 2′-deoxy-2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, a locked nucleotide, an abasic nucleotide, 2′-amino-modified nucleotide, 2′-alkyl-modified nucleotide, morpholino nucleotide, a phosphoramidate, and a non-natural base comprising nucleotide.

18. The dsRNA of claim 14 , comprising at least one 2′-O-methyl modified nucleotide and at least one 2′-deoxythymidine-3′-phosphate nucleotide comprising a 5′-phosphorothioate group.

19. The dsRNA of claim 14 , wherein the sense strand comprises all 2′-O-methyl modified pyrimidines and the antisense strand comprises 2′-O-methyl modified pyrimidines when the pyrimidine is adjacent to A and each strand comprises dTdT at the 3′ end.

20. The dsRNA of claim 14 , wherein the sense strand comprises all 2′-O-methyl modified pyrimidines and the antisense strand comprises 2′-O-methyl modified pyrimidines when the pyrimidine is adjacent to A and each strand comprises dTsdT at the 3′ end.

21. The dsRNA of claim 14 , wherein the sense strand comprises all 2′-O-methyl modified pyrimidines and the antisense strand comprises 2′-O-methyl modified pyrimidines when a) the pyrimidine is adjacent to A or b) the pyrimidine is a uracil adjacent to a U or a G, and each strand comprises dTsdT at the 3′ end.

22. The dsRNA of claim 1 or claim 2 or claim 6 , further comprising a ligand.

23. The dsRNA of claim 22 , wherein the ligand is conjugated to the 3′-end of the sense strand of the dsRNA.

24. A composition for inhibiting expression of a GNAQ gene comprising the dsRNA of claim 1 or claim 2 or claim 6 and pharmaceutical formulation.

25. The composition of claim 24 , wherein the pharmaceutical formulation is a lipid formulation.

26. The composition of claim 24 , wherein the pharmaceutical formulation is a a (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl 4-(dimethylamino)butanoate (MC3) comprising formulation, or a LNP11 formulation.

27. An isolated cell containing the dsRNA of claim 1 or claim 2 or claim 6 .

28. A vector comprising a nucleotide sequence that encodes at least one strand of the dsRNA of claim 1 or claim 6 .

29. An isolated cell comprising the vector of claim 28 .

30. The dsRNA of claim 1 , wherein said dsRNA, upon contact with a cell expressing said GNAQ, inhibits expression of said GNAQ gene by at least 40% compared to a cell not so contacted.

31. The dsRNA of claim 1 , wherein the dsRNA has an IC50 of less than 10 pM.

32. A method of inhibiting GNAQ expression in a cell, the method comprising:

(a) introducing into the cell the dsRNA of claim 1 or claim 2 or claim 6 ; and

(b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of a GNAQ gene, thereby inhibiting expression of the GNAQ gene in the cell.

33. A method of treating a disorder mediated by GNAQ expression comprising administering to a human in need of such treatment a therapeutically effective amount of the dsRNA of claim 1 or claim 2 or claim 6 .

34. The method of claim 33 , wherein the human has uveal melanoma, cutaneous melanoma, Blue nevi, Nevi of Ota, a small lung tumor, or a neuroendocrine tumors.

35. The composition of claim 24 , wherein the pharmaceutical formulation is a (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl 4-(dimethylamino)butanoate (MC3) comprising formulation.

36. The composition of claim 24 , wherein the pharmaceutical formulation is a LNP11 formulation.