Compositions for modulating C9ORF72 expression
Disclosed herein are compositions and methods for reducing expression of C9ORF72 mRNA and protein in an animal with C9ORF72 specific inhibitors. Also disclosed herein are compositions and methods of selectively inhibiting a C9ORF72 pathogenic associated mRNA variant by administering an antisense compound targeting the region beginning at the start site of exon 1A to the start site of exon 1B of a C9ORF72 pre-mRNA. Such methods are useful to treat, prevent, or ameliorate neurodegenerative diseases in an individual in need thereof. Such C9ORF72 specific inhibitors include antisense compounds.
1. A compound comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and having a nucleobase sequence comprising at least 12 consecutive nucleobases of a nucleobase sequence selected from SEQ ID NOs: 442-467.
2. A compound comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8 consecutive nucleobases complementary to an equal length portion of nucleobases 7860-7906 or 7907-7944 of SEQ ID NO: 2.
3. The compound of claim 2 , wherein the modified oligonucleotide is a single-stranded modified oligonucleotide.
4. The compound of claim 3 , wherein at least one internucleoside linkage of the modified oligonucleotide is a modified internucleoside linkage.
5. The compound of claim 4 , wherein the modified internucleoside linkage is a phosphorothioate internucleoside linkage.
6. The compound of claim 4 , wherein each internucleoside linkage of the modified oligonucleotide is a modified internucleoside linkage.
7. The compound of claim 6 , wherein the modified internucleoside linkage is a phosphorothioate linkage.
8. The compound of claim 5 , wherein at least one internucleoside linkage of the modified oligonucleotide is a phosphodiester internucleoside linkage.
9. The compound of claim 2 , wherein at least one nucleobase of the modified oligonucleotide comprises a modified nucleobase.
10. The compound of claim 9 , wherein the modified nucleobase is a 5-methylcytosine.
11. The compound of claim 2 , wherein at least one nucleoside of the modified oligonucleotide comprises a modified sugar.
12. The compound of claim 11 , wherein each nucleoside of the modified oligonucleotide comprises a modified sugar.
13. The compound of claim 11 , wherein the modified sugar is a bicyclic sugar.
14. The compound of claim 13 , wherein each bicyclic sugar comprises a chemical bridge between the 4′ and 2′ positions of the sugar, wherein each chemical bridge is independently selected from: 4′-CH(R)—O-2′ and 4′-(CH 2 ) 2 —O-2′, wherein R is independently selected from H, C 1 -C 6 alkyl, and C 1 -C 6 alkoxy.
15. The compound of claim 14 , wherein at least one chemical bridge is 4′-CH(R)—O-2′ and wherein R is methyl.
16. The compound of claim 14 , wherein at least one chemical bridge is 4′-CH(R)—O-2′ and wherein R is H.
17. The compound of claim 14 , wherein at least one chemical bridge is 4′-CH(R)—O-2′ and wherein R is CH 2 —O—CH 3 .
18. The compound of claim 11 , wherein at least one modified sugar comprises a 2′-O-methoxyethyl group.
19. The compound of claim 11 , wherein at least one modified sugar comprises a 2′-O-methyl group.
20. The compound of claim 12 , wherein each modified sugar comprises a 2′-O-methoxyethyl group or a 2′-O-methyl group.
21. The compound of claim 2 , wherein the modified oligonucleotide is a gapmer.
22. The compound of claim 2 , wherein the modified oligonucleotide comprises:
a gap segment consisting of 10 linked deoxynucleosides;
a 5′ wing segment consisting of 5 linked nucleosides; and
a 3′ wing segment consisting of 5 linked nucleosides;
wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.
23. The compound of claim 2 , wherein the modified oligonucleotide comprises:
a gap segment consisting of 8 linked deoxynucleosides;
a 5′ wing segment consisting of 5 linked nucleosides; and
a 3′ wing segment consisting of 5 linked nucleosides;
wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.
24. The compound of claim 2 , wherein the modified oligonucleotide comprises sugar modified nucleosides in any of the following patterns: eeekkdddddddkkeee, eekkddddddddkkeee, ekddddddddekekeee, kekeddddddddekeke, and ekekddddddddkekee; wherein,
e=a 2′-O-methoxyethyl modified nucleoside,
d=a 2′-deoxynucleoside, and
k=a cEt nucleoside.
25. The compound of claim 2 , wherein the modified oligonucleotide comprises internucleoside linkages in any of the following patterns: soooossssssssssooss, sooosssssssssooss, soosssssssssooss, and sosssssssssoooss; wherein,
s=a phosphorothioate linkage, and
o=a phosphodiester linkage.
26. The compound of claim 2 , wherein the modified oligonucleotide consists of 17, 18, 19, or 20 linked nucleosides.
27. The compound of claim 2 , wherein the modified oligonucleotide has a nucleobase sequence complementary to a region of C9ORF72 other than a hexanucleotide repeat expansion, wherein the hexanucleotide repeat expansion comprises any of GGGCC, GGGGGG, GGGGCG, and GGGGGC.
28. A conjugated antisense compound comprising the compound of claim 2 .
29. The compound of claim 2 , consisting of the modified oligonucleotide.
30. A double-stranded compound comprising the compound of claim 2 .
31. A conjugated antisense compound comprising the double-stranded compound of claim 30 .
32. A composition comprising a compound according to claim 2 and at least one of a pharmaceutically acceptable carrier or diluent, wherein optionally the pharmaceutically acceptable diluent is phosphate-buffered saline (PBS).
33. The composition according to claim 32 , wherein the compound is a salt, wherein optionally the salt is a sodium salt.