2. Academic Validation
  3. A Novel Mouse Model Harboring Hepatitis B Virus Covalently Closed Circular DNA

A Novel Mouse Model Harboring Hepatitis B Virus Covalently Closed Circular DNA

  • Cell Mol Gastroenterol Hepatol. 2022;13(4):1001-1017. doi: 10.1016/j.jcmgh.2021.11.011.
Zaichao Xu 1,  Li Zhao 1,  Youquan Zhong 1,  Chengliang Zhu 2,  Kaitao Zhao 1,  Yan Teng 1,  Xiaoming Cheng 3,  Qiang Chen 4,  Yuchen Xia 5
Affiliations

Affiliations

  • 1 State Key Laboratory of Virology and Hubei Province Key Laboratory of Allergy and Immunology, Institute of Medical Virology, School of Basic Medical Sciences, Wuhan University, Wuhan, China.
  • 2 Department of Clinical Laboratory, Renmin Hospital of Wuhan University, Wuhan, China.
  • 3 State Key Laboratory of Virology and Hubei Province Key Laboratory of Allergy and Immunology, Institute of Medical Virology, School of Basic Medical Sciences, Wuhan University, Wuhan, China; Wuhan University Center for Pathology and Molecular Diagnostics, Zhongnan Hospital of Wuhan University, Wuhan, China; Hubei Clinical Center and Key Laboratory of Intestinal and Colorectal Diseases, Wuhan, China.
  • 4 Department of Radiation and Medical Oncology, Medical Research Institute, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan, China; Frontier Science Center for Immunology and Metabolism, Medical Research Institute, Wuhan University, Wuhan, China.
  • 5 State Key Laboratory of Virology and Hubei Province Key Laboratory of Allergy and Immunology, Institute of Medical Virology, School of Basic Medical Sciences, Wuhan University, Wuhan, China. Electronic address: [email protected].
PMID: 34896285 DOI: 10.1016/j.jcmgh.2021.11.011
Abstract

Background and aims: The persistence of viral covalently closed circular DNA (cccDNA) is the major obstacle for Antiviral treatment against hepatitis B virus (HBV). Basic and translational studies are largely hampered due to the lack of feasible small animal models to support HBV cccDNA formation. The aim of this study is to establish a novel mouse model harboring cccDNA.

Methods: An adeno-associated virus (AAV) vector carrying a replication-deficient HBV1.04-fold genome (AAV-HBV1.04) was constructed. The linear HBV genome starts from nucleotide 403 and ends at 538, which results in the splitting of HBV surface and polymerase genes. Different HBV replication markers were evaluated for AAV-HBV1.04 plasmid-transfected cells, the AAV-HBV1.04 viral vector-transduced cells, and mice injected with the AAV-HBV1.04 viral vector.

Results: Compared with the previously reported AAV-HBV1.2 construct, direct transfection of AAV-HBV1.04 plasmid failed to produce hepatitis B surface antigen and progeny virus. Interestingly, AAV-HBV1.04 viral vector transduction could result in the formation of cccDNA and the production of all HBV replication markers in vitro and in vivo. The formation of cccDNA could be blocked by ATR (ataxia-telangiectasia and Rad3-related protein) inhibitors but not HBV reverse transcription inhibitor or capsid inhibitors. The AAV-HBV1.04 mouse supported long-term HBV replication and responded to Antiviral treatments.

Conclusions: This AAV-HBV1.04 mouse model can support HBV cccDNA formation through ATR-mediated DNA damage response. The de novo formed cccDNA but not the parental AAV vector can lead to the production of hepatitis B surface antigen and HBV progeny. This model will provide a unique platform for studying HBV cccDNA and developing novel antivirals against HBV Infection.

Keywords

Adeno-Associated Virus; Antiviral; HBV; Immune-Competent Mouse; cccDNA.

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