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.

CHDI Foundation CHDI Management Princeton New Jersey

Department of Medicine University of Massachusetts Medical School Worcester Massachusetts

Department of Neurobiology University of Massachusetts Medical School Worcester Massachusetts

Department of Neurosurgery University of Massachusetts Medical School Worcester Massachusetts

Department of Pediatrics University of Massachusetts Medical School Worcester Massachusetts

Department of Physiology Development and Neuroscience University of Cambridge Cambridge United Kingdom

Geisel School of Medicine Dartmouth College Hanover New Hampshire

Horae Gene Therapy Center University of Massachusetts Medical School Worcester Massachusetts

MassGeneral Institute for Neurodegenerative Disease Charlestown Massachusetts

Preclinical Imaging and Research Laboratories South Australian Health and Medical Research Institute Gilles Plains South Australia

Research Center PIGMOD Institute of Animal Physiology and Genetics of the Czech Academy of Sciences Libechov Czech Republic

RNA Therapeutics Institute University of Massachusetts Medical School Worcester Massachusetts

School of Biological Sciences University of Auckland Auckland New Zealand

West China School of Medicine West China Hospital Sichuan University Chengdu China

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Emerging Roles of Exosomes in Huntington's Disease

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