Screening for inhibitor of episomal DNA identified dicumarol as a hepatitis B virus inhibitor

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  • Fumihiko Takeuchi, Kyoto University
  • ,
  • Sotaro Ikeda, Kyoto University
  • ,
  • Yuta Tsukamoto, Kyoto University, University Hospital Bonn
  • ,
  • Yoshikazu Iwasawa, Kyoto University
  • ,
  • Chen Qihao, Kyoto University
  • ,
  • Yukie Otakaki, Kyoto University
  • ,
  • Ouda Ryota, Kyoto University, Hokkaido University
  • ,
  • Wan Ling Yao, Kyoto University
  • ,
  • Ryo Narita
  • Hijikata Makoto, Kyoto University
  • ,
  • Koichi Watashi, National Institute of Infectious Diseases, Tokyo University of Science, Japan Science and Technology Agency
  • ,
  • Takaji Wakita, National Institute of Infectious Diseases
  • ,
  • Koh Takeuchi, National Institute of Advanced Industrial Science and Technology
  • ,
  • Kazuaki Chayama, Hiroshima University
  • ,
  • Amane Kogure, Kyoto University
  • ,
  • Hiroki Kato, Kyoto University, University Hospital Bonn
  • ,
  • Takashi Fujita, Kyoto University

Currently, there is no available therapy to eradicate hepatitis B virus (HBV) in chronically infected individuals. This is due to the difficulty in eliminating viral covalently closed circular (ccc) DNA, which is central to the gene expression and replication of HBV. We developed an assay system for nuclear circular DNA using an integration-deficient lentiviral vector. This vector produced non-integrated circular DNA in nuclei of infected cells. We engineered this vector to encode firefly luciferase to monitor the lentiviral episome DNA. We screened 3,840 chemicals by this assay for luciferase-reducing activity and identified dicumarol, which is known to have anticoagulation activity. We confirmed that dicumarol reduced lentiviral episome DNA. Furthermore, dicumarol inhibited HBV replication in cell culture using NTCP-expressing HepG2 and primary human hepatocytes. Dicumarol reduced intracellular HBV RNA, DNA, supernatant HBV antigens and DNA. We also found that dicumarol reduced the cccDNA level in HBV infected cells, but did not affect HBV adsorption/entry. This is a novel assay system for screening inhibitors targeting nuclear cccDNA and is useful for finding new antiviral substances for HBV.

OriginalsprogEngelsk
Artikelnummere0212233
TidsskriftPLOS ONE
Vol/bind14
Nummer2
ISSN1932-6203
DOI
StatusUdgivet - feb. 2019

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