TY - JOUR
T1 - A novel recombinant cccDNA-based mouse model with long term maintenance of rcccDNA and antigenemia
AU - Wu, Min
AU - Wang, Cong
AU - Shi, Bisheng
AU - Fang, Zhong
AU - Qin, Boyin
AU - Zhou, Xiaohui
AU - Zhang, Xiaonan
AU - Yuan, Zhenghong
N1 - Funding Information:
We thank Prof. Qiang Deng (Fudan University, China) for his generous gift of prcccDNA plasmid and Prof. Xianghui Yu (Jilin University, China) for AAV8 vectors; we thank Ark Biosciences Company (Shanghai, China) for providing AKEX0007; we thank Zhuying Chen for her excellent technical assistance. This study is supported by the Innovation Program of Shanghai Municipal Education Commission ( 2017-01-07-00-07-E00057 ), National Science and Technology Major Project of China ( 2017ZX10302201001005 ), National Natural Science Foundation of China ( 81873962 , 81671998 , 91842309 ), Shanghai Science and Technology Commission ( 16411960100 ), Local Innovative and Research Teams Project of Guangdong Pearl River Talents Program ( 2017BT01S131 ).
Funding Information:
We thank Prof. Qiang Deng (Fudan University, China) for his generous gift of prcccDNA plasmid and Prof. Xianghui Yu (Jilin University, China) for AAV8 vectors; we thank Ark Biosciences Company (Shanghai, China) for providing AKEX0007; we thank Zhuying Chen for her excellent technical assistance. This study is supported by the Innovation Program of Shanghai Municipal Education Commission (2017-01-07-00-07-E00057), National Science and Technology Major Project of China (2017ZX10302201001005), National Natural Science Foundation of China (81873962, 81671998, 91842309), Shanghai Science and Technology Commission (16411960100), Local Innovative and Research Teams Project of Guangdong Pearl River Talents Program (2017BT01S131).
Publisher Copyright:
© 2020
PY - 2020/8
Y1 - 2020/8
N2 - The covalently closed circular DNA (cccDNA) of hepatitis B virus (HBV) is critical for viral persistence in vivo. The lack of reliable, characterized and convenient small animal models for studying cccDNA persistence has long been a bottleneck for basic and translational research on HBV cure. A mouse model that can maintain intrahepatic cccDNA is urgently needed. Through combining the Cre/loxP-mediated recombination and adeno-associated virus (AAV) vector delivery strategy, we establish a novel recombinant cccDNA (rcccDNA) mouse model. AAV-rcccDNA mice supported long-term maintenance of intrahepatic rcccDNA which could be easily detected by Southern blotting within 30 weeks after transduction. Quantitative PCR could detect the rcccDNA signal throughout the experiment duration (>51 weeks). Furthermore, rcccDNA supported persistent serum antigenemia (>72 weeks) and intrahepatic HBsAg and HBcAg expression (>51 weeks). Flow cytometry analysis and single-cell RNA sequencing showed that AAV-rcccDNA mice displayed a compromised CD8+ T cell response. Meanwhile, minimal intrahepatic inflammation and fibrosis were observed. Furthermore, three anti-HBV compounds, AKEX0007, a post-transcriptional inhibitor, Bay 41-4109, a capsid allosteric modulator, and Entecavir were assessed in this AAV-rcccDNA mouse model. The changes of viral markers by these drugs were consistent with their mode of action although neither of them diminished the level of rcccDNA. This mouse model recapitulated the immune tolerant state of HBV infection with long term maintenance of cccDNA and antigenemia, which will provide a suitable platform for studying cccDNA persistence and developing intervention strategies that would eventually break the tolerance and clear the virus.
AB - The covalently closed circular DNA (cccDNA) of hepatitis B virus (HBV) is critical for viral persistence in vivo. The lack of reliable, characterized and convenient small animal models for studying cccDNA persistence has long been a bottleneck for basic and translational research on HBV cure. A mouse model that can maintain intrahepatic cccDNA is urgently needed. Through combining the Cre/loxP-mediated recombination and adeno-associated virus (AAV) vector delivery strategy, we establish a novel recombinant cccDNA (rcccDNA) mouse model. AAV-rcccDNA mice supported long-term maintenance of intrahepatic rcccDNA which could be easily detected by Southern blotting within 30 weeks after transduction. Quantitative PCR could detect the rcccDNA signal throughout the experiment duration (>51 weeks). Furthermore, rcccDNA supported persistent serum antigenemia (>72 weeks) and intrahepatic HBsAg and HBcAg expression (>51 weeks). Flow cytometry analysis and single-cell RNA sequencing showed that AAV-rcccDNA mice displayed a compromised CD8+ T cell response. Meanwhile, minimal intrahepatic inflammation and fibrosis were observed. Furthermore, three anti-HBV compounds, AKEX0007, a post-transcriptional inhibitor, Bay 41-4109, a capsid allosteric modulator, and Entecavir were assessed in this AAV-rcccDNA mouse model. The changes of viral markers by these drugs were consistent with their mode of action although neither of them diminished the level of rcccDNA. This mouse model recapitulated the immune tolerant state of HBV infection with long term maintenance of cccDNA and antigenemia, which will provide a suitable platform for studying cccDNA persistence and developing intervention strategies that would eventually break the tolerance and clear the virus.
KW - Adeno-associated virus (AAV)
KW - Cre/loxP
KW - Hepatitis B virus (HBV)
KW - Mouse model
KW - Recombinant covalently closed circular DNA (rcccDNA)
UR - http://www.scopus.com/inward/record.url?scp=85086605078&partnerID=8YFLogxK
U2 - 10.1016/j.antiviral.2020.104826
DO - 10.1016/j.antiviral.2020.104826
M3 - Article
C2 - 32502604
SN - 1872-9096
VL - 180
SP - 1
EP - 10
JO - Antiviral Research
JF - Antiviral Research
M1 - 104826
ER -
