Hepatitis B virus inhibition in mice by lentiviral vector mediated short hairpin RNA.
BACKGROUND
Chronic hepatitis B virus (HBV) infection is an important cause of cirrhosis and hepatocellular carcinoma. The major challenges for current therapies are the low efficacy of current drugs and the occurrence of drug resistant HBV mutations. RNA interference (RNAi) of virus-specific genes offers the possibility of developing a new anti-HBV therapy. Recent reports have shown that lentiviral vectors based on HIV-1 are promising gene delivery vehicles due to their ability to integrate transgenes into non-dividing cells. Herein, a lentivirus-based RNAi system was developed to drive expression and delivery of HBV-specific short hairpin RNA (shRNA) in a mouse model for HBV replication.


METHODS
Hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) in the sera of the mice were analyzed by quantitative sandwich enzyme linked immunosorbent assay (ELISA) technique, hepatitis B core antigen (HBcAg) and HBsAg in the livers of the mice were detected by immunohistochemical assay, HBV DNA and HBV mRNA were measured by fluorogenic quantitative polymerase chain reaction (FQ-PCR) and quantitative real-time PCR respectively.


RESULTS
Co-injection of HBV plasmids together with the lentivirus targeting HBV shRNA induced an RNAi response. Secreted HBsAg was reduced by 89% in mouse serum, and HBeAg was also significantly inhibited, immunohistochemical detection of HBcAg or HBsAg in the liver tissues also revealed substantial reduction. Lentiviral mediated shRNA caused a significant suppression in the levels of viral mRNA and DNA synthesis compared to the control group.


CONCLUSIONS
Lentivirus-based RNAi can be used to suppress HBV replication in vivo, it might become a potential therapeutic strategy for treating HBV and other viral infections.
Liver Transplantation Center of the First Affiliated Hospital, Nanjing Medical University, 300 Guangzhou Road, Nanjing, Jiangsu 210029, PR China. szwam@nuaa.edu.cn
Lei
L
Deng
Guoqiang
G
Li
Lisen
L
Xi
Aihong
A
Yin
Yun
Y
Gao
Wei
W
You
Xuehao
X
Wang
Beicheng
B
Sun
D000818:Animals; D004279:DNA, Viral / Q000235:genetics; D004195:Disease Models, Animal; D005260:Female; D005822:Genetic Vectors / Q000235:genetics / Q000627:therapeutic use*; D006512:Hepatitis B Core Antigens / Q000378:metabolism; D006514:Hepatitis B Surface Antigens / Q000378:metabolism; D006513:Hepatitis B e Antigens / Q000378:metabolism; D006515:Hepatitis B virus / Q000235:genetics / Q000502:physiology*; D019694:Hepatitis B, Chronic / Q000628:therapy; D016086:Lentivirus / Q000235:genetics*; D008099:Liver / Q000378:metabolism / Q000821:virology; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D034622:RNA Interference / Q000502:physiology*; D012367:RNA, Viral / Q000235:genetics; D014779:Virus Replication / Q000235:genetics / Q000502:physiology*
D004279:DNA, Viral; D006512:Hepatitis B Core Antigens; D006514:Hepatitis B Surface Antigens; D006513:Hepatitis B e Antigens; D012367:RNA, Viral