Treatment of amyotrophic lateral sclerosis
The invention relates to a method for the treatment of amyotrophic lateral sclerosis (ALS). Specifically, the invention implements the use of an antisense sequence adapted to affect alternative splicing in a human SOD1 pre-mRNA, thereby leading to the destruction of the skipped m RNA by the cell machinery.
1. An antisense oligonucleotide of 20 to 30 nucleotides in length targeting a human SOD1 pre-mRNA, wherein said antisense oligonucleotide comprises SEQ ID NO:1.
2. A nucleic acid molecule comprising: (i) a nucleic acid sequence of SEQ ID NO:1 and (ii) a nucleic acid sequence of SEQ ID NO:4.
3. An antisense oligonucleotide of 20 to 30 nucleotides in length targeting a human SOD1 pre-mRNA, wherein said antisense oligonucleotide comprises SEQ ID NO: 1 and wherein said antisense oligonucleotide is modified with a small nuclear RNA such as the U7 small nuclear RNA.
4. A vector comprising a nucleic acid that encodes an antisense oligonucleotide of 20 to 30 nucleotides in length targeting a human SOD1 pre-mRNA, wherein said antisense oligonucleotide comprises SEQ ID NO: 1.
5. The vector according to claim 4 , which is a viral vector.
6. The vector according to claim 5 , wherein said viral vector is an AAV vector, in particular an AAV9 or AAV10 vector.
7. A vector encoding at least one antisense oligonucleotide targeting a human SOD1 pre-mRNA, wherein said antisense oligonucleotide induces exon-skipping in said pre-mRNA, wherein said vector further comprises an expression cassette containing a nucleotide sequence encoding a human SOD1 protein, wherein said nucleotide sequence comprises SEQ ID NO:11 or SEQ ID NO:12, wherein the antisense oligonucleotide cannot induce exon-skipping in the pre-mRNA encoded by said nucleotide sequence.
8. A method for treating amyotrophic lateral sclerosis, comprising administering to a subject in need thereof an antisense oligonucleotide of 20 to 30 nucleotides in length targeting a human SOD1 pre-mRNA, wherein said antisense oligonucleotide comprises SEQ ID NO: 1.
9. The method of claim 8 , wherein said antisense oligonucleotide is administered via the intravenous or intracerebroventricular routes.
10. A nucleic acid sequence comprising the sequence of SEQ ID NO:11 or 12.
11. An expression cassette comprising the nucleic acid sequence of claim 10 .
12. A vector comprising the nucleic acid sequence of claim 10 .
13. The vector according to claim 12 , wherein said vector is a plasmid or a viral vector.
14. A host cell transformed with a vector according to claim 13 .
15. The host cell according to claim 14 , said cell being an eukaryotic or prokaryotic cell.
16. The host cell according to claim 14 , being a mammalian, human or non-human cell.
17. The host cell according to claim 16 , with the proviso that when the cell is a human cell, said cell is not a human embryonic stem cell.
18. The vector according to claim 7 , wherein the antisense oligonucleotide comprises SEQ ID NO:1 or SEQ ID NO:4.
19. The vector according to claim 7 , wherein the vector encodes an antisense oligonucleotide comprising SEQ ID NO:1 and an antisense oligonucleotide comprising SEQ ID NO:4.
20. A method for treating amyotrophic lateral sclerosis, comprising administering to a subject in need thereof the vector according to claim 4 .
21. The method of claim 20 , wherein said vector is administered via the intravenous or intracerebroventricular routes.
22. A method for treating amyotrophic lateral sclerosis, comprising administering to a subject in need thereof the vector according to claim 7 .
23. The method of claim 22 , wherein said vector is administered via the intravenous or intracerebroventricular routes.
24. The nucleic acid molecule according to claim 2 , wherein said nucleic acid sequence is modified with a small nuclear RNA such as the U7 small nuclear RNA.
25. A method for treating amyotrophic lateral sclerosis, comprising administering to a subject in need thereof the nucleic acid molecule according to claim 2 .
26. The method of claim 25 , wherein said nucleic acid molecule is administered via the intravenous or intracerebroventricular routes.
27. A vector comprising the nucleic acid molecule according to claim 2 .
28. The vector according to claim 27 , which is a viral vector.
29. The vector according to claim 28 , wherein said viral vector is an AAV vector, in particular an AAV9 or AAV10 vector.
30. A method for treating amyotrophic lateral sclerosis, comprising administering to a subject in need thereof the vector according to claim 27 .
31. The method of claim 30 , wherein said vector is administered via the intravenous or intracerebroventricular routes.
32. A composition comprising an antisense oligonucleotide comprising SEQ ID NO:1 and an antisense oligonucleotide comprising SEQ ID NO:4.
33. A method for treating amyotrophic lateral sclerosis, comprising administering to a subject in need thereof the composition according to claim 32 .
34. The method of claim 33 , wherein said composition is administered via the intravenous or intracerebroventricular routes.