Inhibition of HBV replication and gene expression in vitro and in vivo with a single AAV vector delivering two shRNA molecules.
Hepatitis B virus (HBV) infection is highly prevalent worldwide. The major challenge for current antiviral treatment is the elevated drug resistance that occurs via rapid viral mutagenesis. In this study, we developed AAV vectors to simultaneously deliver two or three shRNAs targeting different HBV-related genes. These vectors showed markedly better antiviral effects than ones that delivered a single shRNA in vitro. A dual shRNA expression vector (AAV-157i/1694i), which simultaneously expressed two shRNAs targeted the S and X genes of HBV, reduced HBsAg, HBeAg and HBV DNA levels by 87+/-4, 80.3+/-2.6 and 86.2+/- 7% respectively, eight days post-transduction. In a mouse model of prophylactic treatment, HBsAg and HBeAg were reduced to undetectable levels and the serum HBV DNA level was reduced by at least 100 fold. These results indicate that AAV-157i/1694i generates potent anti-HBV effects and that the strategy of constructing multi-shRNA expression vectors may lead to enhanced anti-HBV efficacy and overcome the evading mechanism of the virus and thus the development of drug resistance. [BMB reports 2009; 42(1): 59-64].
Li
Zhi
Z
Department of Microbiology, The Chinese University of Hong Kong, Hong Kong, China.
He
Ming-Liang
ML
Yao
Hong
H
Dong
Qing-Ming
QM
Chen
Yang-Chao
YC
Chan
Chu-Yan
CY
Zheng
Bo-Jian
BJ
Yuen
Kwok-Yung
KY
Peng
Ying
Y
Sun
Qiang
Q
Yang
Xiao
X
Lin
Marie C
MC
Sung
Joseph J Y
JJ
Kung
Hsiang-Fu
HF
D000818:Animals; D002460:Cell Line; D004279:DNA, Viral / Q000235:genetics; D000229:Dependovirus / Q000235:genetics*; D015967:Gene Expression Regulation, Viral / Q000187:drug effects; D005822:Genetic Vectors / Q000235:genetics*; D006514:Hepatitis B Surface Antigens / Q000097:blood / Q000235:genetics; D006513:Hepatitis B e Antigens / Q000097:blood / Q000235:genetics; D006515:Hepatitis B virus / Q000187:drug effects / Q000235:genetics* / Q000502:physiology*; D006801:Humans; D008099:Liver / Q000473:pathology / Q000821:virology; D051379:Mice; D010957:Plasmids / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics* / Q000494:pharmacology; D014779:Virus Replication / Q000187:drug effects
D004279:DNA, Viral; D006514:Hepatitis B Surface Antigens; D006513:Hepatitis B e Antigens; D034741:RNA, Small Interfering