Compositions and methods for modulation of LMNA expression
View Patent ↗Disclosed herein are compounds, compositions and methods for modulating the expression of LMNA in a cell, tissue or animal. Also provided are methods of target validation. Also provided are uses of disclosed compounds and compositions in the manufacture of a medicament for treatment of diseases and disorders. Further provided are methods of identifying cis splicing regulatory elements of a selected mRNA using the disclosed compounds.
1. A method of modulating splicing of a LMNA pre-mRNA in a cell, comprising contacting the cell with a single-stranded antisense oligomeric compound comprising an oligonucleotide consisting of 12 to 30 linked nucleosides, wherein the oligonucleotide has a nucleobase sequence comprising an at least 8 nucleobase portion complementary to an equal length portion of nucleotides 1983-2062 of SEQ ID NO: 4 and is at least 90% complementary to LMNA pre-mRNA having SEQ ID NO:4 as measured over the entirety of the oligonucleotide, wherein the oligonucleotide comprises at least one modified sugar moiety; and
thereby modulating splicing of the LMNA pre-mRNA in the cell, wherein the modulation of splicing results in an increase in the ratio of full-length LMNA mRNA to truncated LMNA mRNA.
2. The method of claim 1 , wherein the modulation of splicing results in an increase in the ratio of full-length lamin A protein to truncated lamin A protein.
3. The method of claim 1 , wherein the modified sugar moiety is a bicyclic nucleic acid selected from the group consisting of 4′-CH 2 —O-2′ BNA or 4′-CH 2 CH 2 —O-2′ BNA.
4. The method of claim 3 , wherein the modified sugar moiety is a 4′-CH 2 —O-2′ BNA.
5. The method of claim 1 , wherein the modified sugar moiety is a 2′-O-(methoxyethyl) modified sugar moiety.
6. The method of claim 1 , wherein the cell is in an animal.
7. The method of claim 1 , wherein the animal is a human.
8. The method of claim 1 , wherein at least one internucleoside linkage of said oligonucleotide is a modified internucleoside linkage.
9. The method of claim 1 , wherein each internucleoside linkage of said oligonucleotide is a phosphorothioate internucleoside linkage.
10. The method of claim 1 , wherein at least one nucleoside of said oligonucleotide comprises a modified nucleobase.
11. The method of claim 10 , wherein the modified nucleobase is a 5-methylcytosine.
12. The method of claim 1 , wherein each cytosine in said oligonucleotide is a 5-methylcytosine.
13. The method of claim 1 , wherein each nucleoside of the oligonucleotide comprises a 2′-O-(methoxyethyl) modified sugar moiety, wherein each internucleoside linkage of said oligonucleotide is a phosphorothioate internucleoside linkage, and wherein each cytosine in said oligonucleotide is a 5-methylcytosine.
14. The method of claim 1 , wherein said oligonucleotide consists of 15 to 30 linked nucleosides.
15. The method of claim 1 , wherein said oligonucleotide consists of 18 to 22 linked nucleosides.
16. The method of claim 1 , wherein each nucleoside of said oligonucleotide comprises the same modified sugar moiety.
17. The method of claim 16 , wherein the modified sugar moiety is a bicyclic nucleic acid selected from the group consisting of 4′-CH 2 —O-2′ BNA or 4′-CH 2 CH 2 —O-2′ BNA.
18. The method of claim 17 , wherein the modified sugar moiety is a 4′-CH 2 —O-2′ BNA.