Hepatitis B virus inhibition in mice by lentiviral vector mediated short hairpin RNA

Abstract

BackgroundChronic 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.MethodsHepatitis 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.ResultsCo-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.ConclusionLentivirus-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 cell-targeted delivery of therapeutic molecules

“…Oligonucleotides (siRNA, microRNA and antisense nucleotide), interferon and peptide aptamers conjugated to synthetic nanoparticles (Chandra et al, 2012;Ryoo et al, 2012), viral vectors and antibodies that can bind to virusinfected hepatocytes (Deng et al, 2009;Ji et al, 2012;Sakurai et al, 2012;Shapira et al, 2012;Yang et al, 2010;Zhang et al, 2009) have been used to suppress virus replication and viral protein synthesis. Viral entry can be blocked by inhibiting the docking of viruses to the surface of hepatocytes using peptides or antibodies (Liu et al, 2012a;Matsumura et al, 2009;Wong-Staal et al, 2010).…”

“…Methods aimed at suppressing virus replication and viral protein synthesis have been developed recently to treat hepatitis (Chandra et al, 2012;Deng et al, 2009;Ji et al, 2012;Ryoo et al, 2012;Sakurai et al, 2012;Yang et al, 2010;Zhang et al, 2009), block virus entry into hepatocytes (Liu et el., 2012a;Matsumura et al, 2009;Wong-Staal et al, 2010) and remove virus-infected cells using cytotoxic agents (Shapira et al, 2012).…”

Expression of CHODL in hepatocellular carcinoma affects invasion and migration of liver cancer cells

“…Chemically synthesized small hairpin RNAs delivered to target cells are known to be transient, lasting only 3–4 days, with low transfection efficiency (16,22). Lentiviral vectors can achieve 70% gene transfer efficacy in both dividing and nondividing hepatocarcinoma cells over a 16-h transduction period with a multiplicity of infection of 1 (4,21,23).…”

Strategies to prevent and reverse liver fibrosis in humans and laboratory animals

“…A lentivirusbased RNAi system was used to deliver HBV-specific short hairpin RNA (shRNA) in a mouse model to suppress HBV replication. This effect was characterized by reduced HBsAg and HBeAg in the serum, suggesting this is a potential therapeutic strategy for treating viral infections (Deng et al 2009;McCaffrey et al 2003).…”