Methods and means for treating DNA repeat instability associated genetic disorders
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 a spino-cerebellar ataxia (SCA), spinal and bulbar muscular atrophy (SBMA), or dentatorubral-pallidoluysian atrophy (DRPLA) in a subject in need thereof, comprising administering to the subject an oligonucleotide comprising or consisting of a sequence that is complementary only to a polyglutamine (CAG)n repetitive nucleotide unit in a gene transcript.
2. The method of claim 1 , wherein the SCA is SCA type 1, 2, 3, 6, 7 or 17.
3. The method of claim 1 , wherein said oligonucleotide has a length of 10 to 50 nucleotides.
4. The method of claim 3 , wherein said oligonucleotide has a length of 12 to 30 nucleotides.
5. The method of claim 4 , wherein said oligonucleotide is a single-stranded oligonucleotide.
6. The method of claim 1 , wherein said oligonucleotide comprises or consists of RNA nucleotides, DNA nucleotides, 2′-O substituted RNA nucleotides, locked nucleic acid (LNA) nucleotides, peptide nucleic acid (PNA) nucleotides, morpholinophosphorodiamidates, ethylene-bridged nucleic acid (ENA) nucleotides or mixtures thereof, with or without a phosphorothioate-containing backbone.
7. The method of claim 6 , wherein the oligonucleotide comprises 2′-O substituted RNA phosphorothioate nucleotides.
8. The method of claim 7 , wherein said 2′-O-substituted RNA phosphorothioate nucleotide is a 2′-O-methyl or 2′-O-methoxy ethyl RNA phosphorothioate nucleotide.
9. The method of claim 1 , wherein said oligonucleotide is provided in an expression vector.
10. The method of claim 9 , wherein said expression vector is a viral vector.
11. The method of claim 1 , wherein said oligonucleotide is provided with an excipient and/or a targeting ligand for delivery of the oligonucleotide to cells and/or for enhancing intracellular delivery of the oligonucleotide.
12. The method of claim 1 , wherein said oligonucleotide is comprised in a pharmaceutically acceptable composition.
13. The method of claim 12 , wherein said pharmaceutical composition further comprises at least one excipient and/or targeting ligand for delivery of the oligonucleotide to the cell and/or for enhancing intracellular delivery of the oligonucleotide.
14. The method of claim 1 , wherein said oligonucleotide preferentially hybridizes to a disease-associated or disease-causing transcript and leaves the function of a normal transcript relatively unaffected.
15. The method of claim 1 , wherein said oligonucleotide prevents the accumulation and/or translation of repeat expanded transcripts in cells.
16. The method of claim 15 , wherein said repeat expanded transcript is a (CAG)n repeat in an ATXN1, ATXN2, ATXN3, SCA7, CACNA1A, AR, SCA17 or DRPLA gene transcript in a cell.
17. The method of claim 1 , wherein said oligonucleotide interferes with gene expression or one or more other precursor RNA or messenger-RNA dependent cellular processes.
18. The method of claim 17 , wherein said messenger-RNA-dependent cellular process is RNA splicing or exon skipping.
19. The method of claim 1 , wherein said administration is carried out by one or more parenteral injections at one or multiple sites in the human body.
20. The method of claim 19 , wherein said parenteral injection is an intravenous, a subcutaneous, an intramuscular, an intrathecal, or an intraventricular injection.
21. A method of treating a spino-cerebellar ataxia (SCA) in a subject in need thereof, comprising administering to the subject the oligonucleotide of claim 1 .
22. A method of treating spinal and bulbar muscular atrophy (SBMA) in a subject in need thereof, comprising administering to the subject the oligonucleotide of claim 1 .
23. A method of treating dentatorubral-pallidoluysian atrophy (DRPLA) in a subject in need thereof, comprising administering to the subject the oligonucleotide of claim 1 .
24. A method of treating a spino-cerebellar ataxia (SCA), spinal and bulbar muscular atrophy (SBMA), or dentatorubral-pallidoluysian atrophy (DRPLA) in a subject in need thereof, comprising administering to the subject an oligonucleotide comprising or consisting of a sequence that is complementary only to a polyglutamine (CAG)n repetitive nucleotide unit in a gene transcript, wherein the oligonucleotide comprises at least one modification and 10 to 50 nucleotides that are complementary to CAGCAGCAGCAG (SEQ ID NO: 18).
25. A method of treating a spino-cerebellar ataxia (SCA), spinal and bulbar muscular atrophy (SBMA), or dentatorubral-pallidoluysian atrophy (DRPLA) in a subject in need thereof, comprising administering to the subject an oligonucleotide comprising or consisting of a sequence that is complementary only to a polyglutamine (CAG)n repetitive nucleotide unit in a gene 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).