Inhibition of hepatitis B virus in mice by RNA interference.
Hepatitis B virus (HBV) infection substantially increases the risk of chronic liver disease and hepatocellular carcinoma in humans. RNA interference (RNAi) of virus-specific genes has emerged as a potential antiviral mechanism. Here we show that RNAi can be applied to inhibit production of HBV replicative intermediates in cell culture and in immunocompetent and immunodeficient mice transfected with an HBV plasmid. Cotransfection with plasmids expressing short hairpin RNAs (shRNAs) homologous to HBV mRNAs induced an RNAi response. Northern and Southern analyses of mouse liver RNA and DNA showed substantially reduced levels of HBV RNAs and replicated HBV genomes upon RNAi treatment. Secreted HBV surface antigen (HBsAg) was reduced by 94.2% in cell culture and 84.5% in mouse serum, whereas immunohistochemical detection of HBV core antigen (HBcAg) revealed >99% reduction in stained hepatocytes upon RNAi treatment. Thus, RNAi effectively inhibited replication initiation in cultured cells and mammalian liver, showing that such an approach could be useful in the treatment of viral diseases.
Department of Pediatrics, Stanford University School of Medicine, 300 Pasteur Drive, Room G305, Stanford, California, USA.
Anton P
AP
McCaffrey
Hiroyuki
H
Nakai
Kusum
K
Pandey
Zan
Z
Huang
Felix H
FH
Salazar
Hui
H
Xu
Stefan F
SF
Wieland
Patricia L
PL
Marion
Mark A
MA
Kay
D000818:Animals; D002460:Cell Line; D005260:Female; D015967:Gene Expression Regulation, Viral / Q000235:genetics; D006512:Hepatitis B Core Antigens / Q000096:biosynthesis / Q000187:drug effects; D006515:Hepatitis B virus / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development*; D008099:Liver / Q000187:drug effects / Q000821:virology; D051379:Mice; D008807:Mice, Inbred BALB C / Q000235:genetics; D008810:Mice, Inbred C57BL / Q000235:genetics; D016688:Mice, Inbred NOD / Q000235:genetics; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000494:pharmacology*; D012367:RNA, Viral / Q000494:pharmacology*; D014779:Virus Replication / Q000187:drug effects* / Q000235:genetics
D006512:Hepatitis B Core Antigens; D012330:RNA, Double-Stranded; D012367:RNA, Viral