Preventing formation of toxic N-terminal huntingtin fragments through antisense oligonucleotide-mediated protein modification
Autor: | Gert-Jan B. van Ommen, Willeke M. C. van Roon-Mom, Annemieke Aartsma-Rus, Melvin M. Evers, Ioannis Zalachoras, Hoang-Dai Tran, Onno C. Meijer, Johan T. den Dunnen |
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Rok vydání: | 2014 |
Předmět: |
Male
congenital hereditary and neonatal diseases and abnormalities Huntingtin Mutant Nerve Tissue Proteins Biology Cleavage (embryo) Biochemistry Cell Line Exon Mice mental disorders Drug Discovery Genetics Huntingtin Protein Animals Humans Molecular Biology chemistry.chemical_classification Serotonin Plasma Membrane Transport Proteins Caspase 6 Exons Oligonucleotides Antisense Molecular biology Exon skipping Peptide Fragments nervous system diseases Amino acid Mice Inbred C57BL Huntington Disease nervous system chemistry Mutation Molecular Medicine Mutant Proteins Trinucleotide repeat expansion Trinucleotide Repeat Expansion Protein Modification Translational Targeted Gene Repair |
Zdroj: | Nucleic Acid Therapeutics Nucleic Acid Therapeutics, 24(1), 4-12 |
ISSN: | 2159-3345 |
Popis: | Huntington's disease (HD) is a progressive autosomal dominant disorder, caused by a CAG repeat expansion in the HTT gene, which results in expansion of a polyglutamine stretch at the N-terminal end of the huntingtin protein. Several studies have implicated the importance of proteolytic cleavage of mutant huntingtin in HD pathogenesis and it is generally accepted that N-terminal huntingtin fragments are more toxic than full-length protein. Important cleavage sites are encoded by exon 12 of HTT. Here we report proof of concept using antisense oligonucleotides to induce skipping of exon 12 in huntingtin pre-mRNA, thereby preventing the formation of a 586 amino acid N-terminal huntingtin fragment implicated in HD toxicity. In vitro studies showed successful exon skipping and appearance of a shorter huntingtin protein. Cleavage assays showed reduced 586 amino acid N-terminal huntingtin fragments in the treated samples. In vivo studies revealed exon skipping after a single injection of antisense oligonucleotides in the mouse striatum. Recent advances to inhibit the formation of mutant huntingtin using oligonucleotides seem promising therapeutic strategies for HD. Nevertheless, huntingtin is an essential protein and total removal has been shown to result in progressive neurodegeneration in vivo. Our proof of concept shows a completely novel approach to reduce mutant huntingtin toxicity not by reducing its expressing levels, but by modifying the huntingtin protein. |
Databáze: | OpenAIRE |
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