Minimal-length Synthetic shRNAs Formulated with Lipid Nanoparticles are Potent Inhibitors of Hepatitis C Virus IRES-linked Gene Expression in Mice

• Published in: Molecular therapy. Nucleic acids Vol. 2; no. 9; p. e123
• Main Authors: Dallas, Anne, Ilves, Heini, Shorenstein, Joshua, Judge, Adam, Spitler, Ryan, Contag, Christopher, Wong, Suet Ping, Harbottle, Richard P, MacLachlan, Ian, Johnston, Brian H
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 2013; Elsevier Limited; Nature Publishing Group; Elsevier
• Subjects: HCV; lipid nanoparticles; Original; RNAi; shRNA; sshRNA; lipid nanoparticles; RNAi; HCV; PK; shRNA; sshRNA
• ISSN: 2162-2531; 2162-2531
• DOI: 10.1038/mtna.2013.50

We previously identified short synthetic shRNAs (sshRNAs) that target a conserved hepatitis C virus (HCV) sequence within the internal ribosome entry site (IRES) of HCV and potently inhibit HCV IRES-linked gene expression. To assess in vivo liver delivery and activity, the HCV-directed sshRNA, SG220 was formulated into lipid nanoparticles (LNP) and injected i.v. into mice whose livers supported stable HCV IRES-luciferase expression from a liver-specific promoter. After a single injection, RNase protection assays for the sshRNA and 3H labeling of a lipid component of the nanoparticles showed efficient liver uptake of both components and long-lasting survival of a significant fraction of the sshRNA in the liver. In vivo imaging showed a dose-dependent inhibition of luciferase expression (>90% 1 day after injection of 2.5 mg/kg sshRNA) with t1/2 for recovery of about 3 weeks. These results demonstrate the ability of moderate levels of i.v.-injected, LNP-formulated sshRNAs to be taken up by liver hepatocytes at a level sufficient to substantially suppress gene expression. Suppression is rapid and durable, suggesting that sshRNAs may have promise as therapeutic agents for liver indications.