Abstract
BackgroundThe sodium taurocholate cotransporting polypeptide (NTCP) is a functional receptor for hepatitis B virus (HBV). NTCP-reconstituted human hepatoma cells support HBV infection, but the infection is suboptimal and no apparent HBV spread has been observed in this system.ResultsWe found that NTCP-reconstituted HepG2 cells were highly susceptible to HBV infection after cells were cultured in a commercial human inducible pluripotent stem cell (iPSC)-derived hepatocyte maintenance medium (HMM). The enhanced HBV infection coincided with increased NTCP expression, and was observed in six different clones of HepG2-NTCP cells. Promoter assays indicated that HMM activated the cytomegalovirus immediate-early (IE) promoter that drives the NTCP expression in the HepG2-NTCP cells. RNA-Seq analysis revealed that HMM upregulated multiple metabolic pathways. Despite highly upregulated NTCP expression by HMM, no obvious HBV spread was observed even in the presence of PEG 8000.ConclusionsOur data suggest that this particular medium could be used to enhance HBV infection in NTCP-reconstituted hepatocytes in vitro.
Highlights
Hepatitis B virus (HBV) infection leads to a wide spectrum of liver diseases ranging from chronic hepatitis, cirrhosis, and hepatocellular carcinoma over several decades [1]
HMM enhances hepatitis B virus (HBV) infection at the entry step In a process of evaluating the utility of induced pluripotent cell-derived hepatocyte-like cells (HLCs) in studying HBV infection, we serendipitously found that a commercial hepatocyte maintenance medium (HMM) designed to maintain inducible pluripotent stem cell (iPSC)-HLCs appeared to enhance HBV infection of HepG2-NTCP cells when compared to the regular medium D10/2%DMSO (DMEM supplemented with 10% fetal bovine serum (FBS) and 2% DMSO; Fig. 1A)
HBV Infection of these cells strictly depended on the transgenic NTCP, as adding during virus inoculation with cyclosporin A, a known HBV entry inhibitor targeting NTCP [37,38,39], resulted in a great reduction in the number of infected cells
Summary
Hepatitis B virus (HBV) infection leads to a wide spectrum of liver diseases ranging from chronic hepatitis, cirrhosis, and hepatocellular carcinoma over several decades [1]. The chronic hepatitis B prevalence was estimated around 3.5% worldwide in 2016, and a total of 257 million people are chronically infected with HBV [2]. HBV is an enveloped virus and a member of Hepadnaviridae family. It contains a partially double-stranded relaxed circular DNA (rcDNA) genome of approximate 3200 bp in length [7]. HBV rcDNA is transported to the nucleus and converted to a covalently closed circular DNA (cccDNA) [8, 9]. The sodium taurocholate cotransporting polypeptide (NTCP) is a functional receptor for hepatitis B virus (HBV). NTCP-reconstituted human hepatoma cells support HBV infection, but the infection is suboptimal and no apparent HBV spread has been observed in this system
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