Study of multiple genetic variations caused by persistent hepatitis C virus replication in long-term cell culture

Nobuyuki Kato, Youki Ueda, Hiroe Sejima, Weilin Gu, Shinya Satoh, Hiromichi Dansako, Masanori Ikeda, Kunitada Shimotohno

Research output: Contribution to journalArticle

Abstract

The most characteristic feature of the hepatitis C virus (HCV) genome in patients with chronic hepatitis C is its remarkable variability and diversity. To better understand this feature, we performed genetic analysis of HCV replicons recovered from two human hepatoma HuH-7-derived cell lines after 1, 3, 5, 7, and 9 years in culture: The cell lines 50-1 and sO harbored HCV 1B-1 and O strain-derived HCV replicons established in 2002 and 2003, respectively. The results revealed that genetic variations in both replicons accumulated in a time-dependent manner at a constant rate despite the maintenance of moderate diversity (less than 1.8% difference) between the clones and that the mutation rate in the 50-1 and sO replicons was 2.5 and 2.9 × 10−3 base substitutions/site/year, respectively. We found that the genetic distance of both replicons increased from 7.9% to 10.5% after 9 years in culture. In addition, we observed that the guanine + cytosine (GC) content of both replicon RNAs increased in a time-dependent manner, as observed in our previous studies. Finally, we demonstrated that the high sensitivity of both replicons to direct-acting antivirals was maintained even after 9 years in culture. Our results suggest that long-term cultured HCV replicon-harboring cells are a useful model for understanding the variability and diversity of the HCV genome and the drug sensitivity of HCV in patients with chronic hepatitis C.

Original languageEnglish
JournalArchives of Virology
DOIs
Publication statusAccepted/In press - Jan 1 2019

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Replicon
Virus Replication
Hepacivirus
Cell Culture Techniques
Chronic Hepatitis C
Genome
Cell Line
Base Composition
Mutation Rate
Antiviral Agents
Hepatocellular Carcinoma
Clone Cells
Maintenance
RNA

ASJC Scopus subject areas

  • Virology

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Study of multiple genetic variations caused by persistent hepatitis C virus replication in long-term cell culture. / Kato, Nobuyuki; Ueda, Youki; Sejima, Hiroe; Gu, Weilin; Satoh, Shinya; Dansako, Hiromichi; Ikeda, Masanori; Shimotohno, Kunitada.

In: Archives of Virology, 01.01.2019.

Research output: Contribution to journalArticle

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