Research Papers: Immunology:
Development of hepatoma-derived, bidirectional oval-like cells as a model to study host interactions with hepatitis C virus during differentiation
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Masahiko Ito1, Suofeng Sun1, Takasuke Fukuhara2, Ryosuke Suzuki3, Miho Tamai4, Toyohiko Yamauchi5, Kenji Nakashima1, Yoh-ichi Tagawa4, Shigetoshi Okazaki6, Yoshiharu Matsuura2, Takaji Wakita3 and Tetsuro Suzuki1
1 Department of Virology and Parasitology, Hamamatsu University School of Medicine, Shizuoka, Japan
2 Department of Molecular Virology, Research Institute for Microbial Diseases, Osaka University, Osaka, Japan
3 Department of Virology II, National Institute of Infectious Diseases, Tokyo, Japan
4 School of Life Science and Technology, Tokyo Institute of Technology, Kanagawa, Japan
5 Central Research Laboratory, Hamamatsu Photonics K.K., Shizuoka, Japan
6 Department of Medical Spectroscopy, Hamamatsu University School of Medicine, Shizuoka, Japan
Tetsuro Suzuki, email:
Keywords: hepatitis C virus, epigenetic reprogramming, hepatoma cell, oval cell, miR200a, Immunology and Microbiology Section, Immune response, Immunity
Received: May 16, 2017 Accepted: June 28, 2017 Published: July 08, 2017
Directed differentiation of human stem cells including induced pluripotent stem cells into hepatic cells potentially leads to acquired susceptibility to hepatitis C virus (HCV) infection. However, cellular determinants that change their expression during cell reprogramming or hepatic differentiation and are pivotal for supporting the HCV life cycle remain unclear. In this study, by introducing a set of reprogramming factors, we established HuH-7-derived oval-like cell lines, Hdo-17 and -23, which possess features of bipotential liver precursors. Upon induction of hepatocyte differentiation, expression of mature hepatocyte markers and hepatoblast markers in cells increased and decreased, respectively. In contrast, in response to cholangiocytic differentiation induction, gene expression of epithelium markers increased and cells formed round cysts with a central luminal space. Hdo cells lost their susceptibility to HCV infection and viral RNA replication. Hepatic differentiation of Hdo cells potentially led to recovery of permissiveness to HCV RNA replication. Gene expression profiling showed that most host-cell factors known to be involved in the HCV life cycle, except CD81, are expressed in Hdo cells comparable to HuH-7 cells. HCV pseudoparticle infectivity was significantly but partially recovered by ectopic expression of CD81, suggesting possible involvement of additional unidentified factors in HCV entry. In addition, we identified miR200a-3p, which is highly expressed in Hdo cells and stem cells but poorly expressed in differentiated cells and mature hepatocytes, as a novel negative regulator of HCV replication. In conclusion, our results showed that epigenetic reprogramming of human hepatoma cells potentially changes their permissivity to HCV.
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