title: Therapeutic inhibition of hepatitis B virus surface antigen expression by RNA interference.
abstract: RNA interference (RNAi) mediated inhibition of virus-specific genes has emerged as a potential therapeutic strategy against virus induced diseases. Human hepatitis B virus (HBV) surface antigen (HBsAg) has proven to be a significant risk factor in HBV induced liver diseases, and an increasing number of mutations in HBsAg are known to enhance the difficulty in therapeutic interventions. The key challenge for achieving effective gene silencing in particular for the purpose of the therapeutics is primarily based on the effectiveness and specificity of the RNAi targeting sequence. To explore the therapeutic potential of RNAi on HBV induced diseases in particular resulted from aberrant or persistent expression of HBsAg, we have especially screened and identified the most potent and specific RNAi targeting sequence that directly mediated inhibition of the HBsAg expression. Using an effective DNA vector-based shRNA expression system, we have screened 10 RNAi targeting sequences (HBsAg-1 to 10) that were chosen from HBsAg coding region, in particular the major S region, and have identified four targeting sequences that could mediate sequence specific inhibition of the HBsAg expression. Among these four shRNAs, an extremely potent and highly sequence specific HBsAg-3 shRNA was found to inhibit HBsAg expression in mouse HBV model. The inhibition was not only preventive in cotransfection experiments, but also had therapeutic effect as assessed by post-treatment protocols. Moreover, this HBsAg-3 shRNA also exhibited a great potency of inhibition in transgenic mice that constitutively expressed HBsAg. These results indicate that HBsAg-3 shRNA can be considered as a powerful therapeutic agent on HBsAg induced diseases.
authors.affiliation: Institute of Basic Medical Sciences, National Cheng Kung University Medical College, 1, University Road, Tainan 701, Taiwan, ROC.
authors.forename: Tsung-Lin
authors.initials: TL
authors.lastname: Cheng
authors.forename: Wen-Wei
authors.initials: WW
authors.lastname: Chang
authors.forename: Ih-Jen
authors.initials: IJ
authors.lastname: Su
authors.forename: Ming-Derg
authors.initials: MD
authors.lastname: Lai
authors.forename: Wenya
authors.initials: W
authors.lastname: Huang
authors.forename: Huan-Yao
authors.initials: HY
authors.lastname: Lei
authors.forename: Wen-Tsan
authors.initials: WT
authors.lastname: Chang
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D006509:Hepatitis B / Q000628:therapy*; D006514:Hepatitis B Surface Antigens / Q000235:genetics* / Q000378:metabolism; D006515:Hepatitis B virus / Q000235:genetics / Q000502:physiology; D051379:Mice; D008807:Mice, Inbred BALB C; D008822:Mice, Transgenic; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000737:chemistry; D014158:Transcription, Genetic; D014779:Virus Replication
chemical_list: D006514:Hepatitis B Surface Antigens; D034741:RNA, Small Interfering