Safe, Long-term Hepatic Expression of Anti-HCV shRNA in a Nonhuman Primate Model

Autor: Romuald Corbau, Olga Zelenaia, Katherine A. High, Sterghios Moschos, Helen Lavender, J. Fraser Wright, David Suhy, Bernd Hauck, Kyle Hayes, Shih-Chu Kao, Laurence O. Whiteley, Tin Mao, B E Souberbielle, Curt Scribner, Andrew D. Burdick, Hubert Denise, Sara H Renison, Alexander A. Kolykhalov, Shangzhen Zhou, Kevin Brady, Peter W. Roelvink
Rok vydání: 2012
Předmět:
Zdroj: Molecular Therapy. 20(9):1737-1749
ISSN: 1525-0016
1525-0024
DOI: 10.1038/mt.2012.119
Popis: The hepatitis C virus (HCV) chronically infects 2% of the world population and effective treatment is limited by long duration and significant side-effects. Here, we describe a novel drug, intended as a “single-shot ” therapy, which expresses three short hairpin RNAs (shRNAs) that simultaneously target multiple conserved regions of the HCV genome as confirmed in vitro by knockdown of an HCV replicon system. Using a recombinant adeno-associated virus (AAV) serotype 8 vector for delivery, comprehensive transduction of hepatocytes was achieved in vivo in a nonhuman primate (NHP) model following a single intravenous injection. However, dose ranging studies performed in 13 NHP resulted in high-expression levels of shRNA from wild-type (wt) Pol III promoters and dose-dependent hepatocellular toxicity, the first demonstration of shRNA-related toxicity in primates, establishing that the hepatotoxicity arises from highly conserved features of the RNA interference (RNAi) pathway. In the second generation drug, each promoter was re-engineered to reduce shRNA transcription to levels that circumvent toxicity but still inhibit replicon activity. In vivo testing of this modified construct in 18 NHPs showed conservation of hepatocyte transduction but complete elimination of hepatotoxicity, even with sustained shRNA expression for 50 days. These data support progression to a clinical study for treatment of HCV infection.
Databáze: OpenAIRE
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