Huntington's disease (HD) is a fatal neurodegenerative disease caused by a genetic expansion of the CAG repeat region in the huntingtin (HTT) gene. Studies in HD mouse models have shown that artificial miRNAs can reduce mutant HTT, but evidence for their effectiveness and safety in larger animals is lacking. HD transgenic sheep express the full-length human HTT with 73 CAG repeats. AAV9 was used to deliver unilaterally to HD sheep striatum an artificial miRNA targeting exon 48 of the human HTT mRNA under control of two alternative promoters: U6 or CβA. The treatment reduced human mutant (m) HTT mRNA and protein 50-80% in the striatum at 1 and 6 months post injection. Silencing was detectable in both the caudate and putamen. Levels of endogenous sheep HTT protein were not affected. There was no significant loss of neurons labeled by DARPP32 or NeuN at 6 months after treatment, and Iba1-positive microglia were detected at control levels. It is concluded that safe and effective silencing of human mHTT protein can be achieved and sustained in a large-animal brain by direct delivery of an AAV carrying an artificial miRNA.
- MeSH
- Dependovirus genetika MeSH
- elektrolyty metabolismus MeSH
- genetické vektory metabolismus MeSH
- geneticky modifikovaná zvířata MeSH
- genom virový MeSH
- Huntingtonova nemoc genetika patologie MeSH
- imunoanalýza MeSH
- injekce MeSH
- játra patofyziologie MeSH
- ledviny patofyziologie MeSH
- lidé MeSH
- messenger RNA genetika metabolismus MeSH
- mikro RNA genetika metabolismus MeSH
- mikroglie metabolismus MeSH
- modely nemocí na zvířatech MeSH
- mutantní proteiny metabolismus MeSH
- neostriatum metabolismus MeSH
- neurony metabolismus MeSH
- ovce MeSH
- protein huntingtin metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
