Treatment of genetic disorders associated with DNA repeat instability
The current invention provides for methods and medicaments that apply oligonucleotide molecules complementary only to a repetitive sequence in a human gene transcript, for the manufacture of a medicament for the diagnosis, treatment or prevention of a cis-element repeat instability associated genetic disorders in humans. The invention hence provides a method of treatment for cis-element repeat instability associated genetic disorders. The invention also pertains to modified oligonucleotides which can be applied in method of the invention to prevent the accumulation and/or translation of repeat expanded transcripts in cells.
1. A method of treating in a subject having a genetic disorder associated with human cis-element repeat instability comprising administering to the subject an oligonucleotide comprising 10 to 50 nucleotides that is complementary to a sequence of a pre-mRNA transcript, said cis-element repeat instability is characterized by CAG repeat instability, said sequence comprising (CAGCAGCAGCAG) SEQ ID NO: 18, said oligonucleotide comprises either ribonucleotides or deoxyribonucleotides, said oligonucleotide comprising a modification selected from the group consisting of: a morpholino phosphorodiamidate oligonucleotide, a phosphorothioate oligonucleotide, a locked nucleic acid (LNA), a peptide nucleic acid (PNA) and an ethylene-bridged nucleic acid, wherein said administered oligonucleotide does not induce RNaseH-mediated degradation of said pre-mRNA transcripts.
2. The method according to claim 1 , wherein said sequence is present in a coding sequence of the gene pre-mRNA transcript.
3. The method according to claim 1 , wherein said sequence is present in a non-coding sequence of the gene pre-mRNA transcript.
4. The method according to claim 1 , wherein the disorder is Huntington's disease.
5. The method according to claim 1 , wherein the oligonucleotide comprises a 2′-O-substituted phosphorothioate nucleotide.
6. The method of claim 5 , wherein said oligonucleotide comprises a 2′-O-methyl phosphorothioate nucleotide or a 2′-O-methyoxy ethyl phosphorothioate nucleotide.
7. A method for reducing the number of CAG repeat-containing gene transcripts in a cell comprising providing to said cell an oligonucleotide comprising a 10 to 50 nucleotides that is complementary to a sequence of a pre-mRNA transcript, said sequence comprising (CAGCAGCAGCAG) SEQ ID NO: 18, said oligonucleotide comprises either ribonucleotides or deoxyribonucleotides, said oligonucleotide comprising a modification selected from the group consisting of: a morpholino phosphorodiamidate oligonucleotide, a phosphorothioate oligonucleotide, a locked nucleic acid (LNA), a peptide nucleic acid (PNA) and an ethylene-bridged nucleic acid, wherein said oligonucleotide does not induce RNaseH-mediated degradation of said pre-mRNA transcripts.
8. The method according to claim 1 , wherein the oligonucleotide has a length selected from the group consisting of 12 to 30 nucleotides, 12 to 45 nucleotides and 12 to 25 nucleotides.
9. The method of claim 7 , wherein the oligonucleotide has a length selected from the group consisting of 12 to 30 nucleotides, 12 to 45 nucleotides and 12 to 25 nucleotides.
10. The method of claim 1 , wherein the oligonucleotide is fully complementary to said sequence.
11. The method of claim 1 , wherein the oligonucleotide is a 2′-O-methyl phosphorothioate.
12. A method of treating a subject having a genetic disorder associated with human cis-element CAG repeat instability comprising administering to the subject an oligonucleotide comprising 10 to 50 nucleotides that is completely complementary to a sequence of a pre-mRNA transcript, wherein the oligonucleotide comprises a sequence selected from the group consisting of: SEQ ID NOs: 5 (cug cug cug cug cug cug cug) and 20 (cug cug cug cug).
13. The method of claim 12 , wherein the oligonucleotide comprises a modification selected from the group consisting of: a morpholino phosphorodiamidate oligonucleotide, a phosphorothioate oligonucleotide, a locked nucleic acid (LNA), a peptide nucleic acid (PNA) and an ethylene-bridged nucleic acid.
14. The method of claim 1 , wherein the oligonucleotide is a RNA phosphorothioate comprising the sequence of SEQ ID NO: 5 (cug cug cug cug cug cug cug) or 20 (cug cug cug cug).
15. The method of claim 1 , wherein the oligonucleotide is a DNA phosphorothioate oligonucleotide.
16. The method of claim 12 , wherein the oligonucleotide is RNA phosphorothioate consisting of the sequence of SEQ ID NO: 5 (cug cug cug cug cug cug cug) or 20 (cug cug cug cug).
17. The method of claim 12 , wherein the oligonucleotide consists of the sequence of SEQ ID NOs: 5 (cug cug cug cug cug cug cug) or 20 (cug cug cug cug).