Compositions and methods of treating amyotrophic lateral sclerosis (ALS)
The present invention relates to small interfering RNA (siRNA) molecules against the SOD1 gene, adeno-associated viral (AAV) vectors encoding siRNA molecules and methods for treating amyotrophic lateral sclerosis (ALS) using the siRNA molecules and AAV vectors.
1. An adeno-associated viral (AAV) vector genome comprising a nucleic acid sequence positioned between two inverted terminal repeats (ITRs); wherein said nucleic acid sequence encodes a sense strand sequence and an antisense strand sequence of an siRNA duplex; wherein the sense strand sequence comprises nucleotides 1-18 of SEQ ID NO. 51; and wherein the antisense strand sequence comprises nucleotides 1-19 of SEQ ID NO. 220.
2. The AAV vector genome of claim 1 , wherein the sense strand sequence and the antisense strand sequence are, independently, 22 nucleotides or less in length.
3. The AAV vector genome of claim 2 , wherein at least one of the sense strand sequence and the antisense strand sequence comprise a 3′ overhang of at least 1 nucleotide.
4. The AAV vector genome of claim 2 , wherein at least one of the sense strand sequence and the antisense strand sequence comprise a 3′ overhang of at least 2 nucleotides.
5. An AAV particle comprising the AAV vector genome of claim 2 .
6. A method for inhibiting the expression of SOD1 gene in a cell comprising administering to the cell a composition comprising an AAV vector genome of claim 2 .
7. The method of claim 6 , wherein the cell is a mammalian cell.
8. The method of claim 7 , wherein the mammalian cell is a motor neuron.
9. The method of claim 7 , wherein the mammalian cell is an astrocyte.
10. A method for treating amyotrophic lateral sclerosis (ALS) caused by SOD1 mutation in a subject, the method comprising administering to the subject a therapeutically effective amount of a composition comprising the AAV particle of claim 5 .
11. The method of claim 10 , wherein the expression of SOD1 mRNA is inhibited or suppressed by up to 93%.
12. The method of claim 10 , wherein the expression of SOD1 mRNA is inhibited or suppressed by about 20% to about 93%.
13. The method of claim 10 , wherein the expression of SOD1 mRNA is inhibited or suppressed by about 50% to about 93%.
14. The method of claim 10 , wherein the ALS is familial ALS with an identified SOD1 gene mutation.
15. The method of claim 10 , wherein the ALS is sporadic ALS caused by SOD1 mutation.
16. The method of claim 10 , wherein the SOD1 gene embraces a mutation that causes a gain of function effect inside the cell.
17. The method of claim 16 , wherein the administration of the composition comprises intraparenchymal spinal administration.
18. The method of claim 10 , wherein the administration of the composition comprises intraparenchymal spinal administration.
19. The AAV vector genome of claim 2 , wherein the sense strand sequence and the antisense strand sequence are, independently, 20 nucleotides in length.
20. The AAV vector genome of claim 2 , wherein the sense strand sequence and the antisense strand sequence are, independently, 21 nucleotides in length.
21. The AAV vector genome of claim 2 , wherein the sense strand sequence and the antisense strand sequence are, independently, 22 nucleotides in length.
22. An siRNA duplex comprising a sense strand sequence and an antisense strand sequence; wherein the sense strand sequence comprises nucleotides 1-18 of SEQ ID NO. 51; and wherein the antisense strand sequence comprises nucleotides 1-19 of SEQ ID NO. 220.
23. The siRNA duplex of claim 22 , wherein the sense strand sequence and the antisense strand sequence are, independently, 20 nucleotides in length.
24. The siRNA duplex of claim 22 , wherein the sense strand sequence and the antisense strand sequence are, independently, 21 nucleotides in length.
25. The siRNA duplex of claim 22 , wherein the sense strand sequence and the antisense strand sequence are, independently, 22 nucleotides in length.
26. The siRNA duplex of claim 22 , wherein at least one of the sense strand sequence and the antisense strand sequence comprise a 3′ overhang of at least 1 nucleotide.
27. The siRNA duplex of claim 22 , wherein at least one of the sense strand sequence and the antisense strand sequence comprise a 3′ overhang of at least 2 nucleotides.