Adeno-Associated Virus Vector Mediated Delivery of the HBV Genome Induces Chronic Hepatitis B Virus Infection and Liver Fibrosis in Mice

Abstract

Liver cirrhosis and hepatocellular carcinomas are major health problems of chronic hepatitis B virus (HBV) infection. To date, rare model has reproduced liver fibrosis associated with long-term HBV infection which in turn has hindered both the understanding of HBV biology and the development of new treatment options. Here, using adeno-associated virus serotype 8 (AAV8) mediated delivery of a 1.2-kb HBV genome, we successfully generated a chronic HBV infectious mouse model that presents the associated liver fibrosis observed following human infection. After AAV8/HBV1.2 vector administration, mice demonstrated effective HBV replication and transcription which resulted in HBV antigen expression and viremia over 6 months. Although no obvious acute inflammatory response was noted, these mice still developed chronic liver disease and hepatic fibrogenesis as demonstrated by increased ground glass-like hepatocytes, an increasing trend of collagen deposition and upregulated fibrosis markers, including type I collagen, type III collagen, tissue inhibitor of metalloproteinase (TIMP), and transforming growth factor-β1(TGF-β1). Taken together, AAV-mediated HBV gene delivery to the mouse liver, induced HBV persistent infection accompanied by liver fibrosis which can serve as a model for investigating the precise mechanisms underlying liver fibrosis following chronic HBV infection as well as for the potential development of novel therapeutics.

Molecular cloning, characterization and expression analysis of TGF-β and receptor genes in the woodchuck model

“…In this study, upregulation of TGF-β1 and TGFBR2 expression in the liver of woodchucks with chronic WHV infection compared to naive controls was observed. This is consistent with the previous finding in humans that CHB patients had higher serum TGF-β1 concentrations (44,45) and increased liver TGFBR2 expression (44)(45)(46)(47)(48)compared to that in the healthy controls (42). Moreover, acute WHV infected woodchucks showed significantly higher TGF-β1 and TGF-β2 expression in the liver than that in the chronically infected woodchucks.…”

The evaluation of fibrotic effects of the hepatitis B virus pre-core in hepatic stellate cells

“…Similarly, prolonged infection of HBV in a mouse model may consequently develop fibrosis in liver (20). Hepatic HSCs accounted as the most important components in the establishment and progression of fibrosis.…”

Hepatitis B Virus X Protein Increases 8-Oxo-7,8-Dihydro-2ʹ-Deoxyguanosine (8-Oxodg) Level via Repressing MTH1/ MTH2 Expression in Hepatocytes

“…They were kept with a 12 h fluorescent dark/light cycle at 24 ± 2°C temperature and 50 ± 5% relative humidity, given a normal laboratory diet and free access to tap water. The plasmid pAAV/HBV1.2 containing a 1.2-kb HBV genome [27] was dissolved in 2 ml physiological saline and injected into the tail veins of mice within 5 seconds following the hydrodynamics-based transfection protocol. Negative control mice were injected with physiological saline only.…”

“…1B). To extrapolate the in vitro result, an HBV infectious mouse model was generated by hydrodynamic injection of a liver-targeted pAAV/HBV1.2 containing a 1.2-kb HBV genome into the tail veins of male C57BL/6 mice [27]. Three days after injection, the amount of 8-oxodG in the liver tissues and serum were measured that demonstrated a higher level in the HBV infectious mice than in the control mice (Fig.…”