Initial Hepatitis C Virus Infection of Adult Hepatocytes Triggers a Temporally Structured Transcriptional Program Containing Diverse Pro- and Antiviral Elements

• Published in: Journal of virology Vol. 95; no. 10
• Main Authors: Tegtmeyer, Birthe, Vieyres, Gabrielle, Todt, Daniel, Lauber, Chris, Ginkel, Corinne, Engelmann, Michael, Herrmann, Maike, Pfaller, Christian K, Vondran, Florian W. R, Broering, Ruth, Vafadarnejad, Ehsan, Saliba, Antoine-Emmanuel, Puff, Christina, Baumgärtner, Wolfgang, Miskey, Csaba, Ivics, Zoltán, Steinmann, Eike, Pietschmann, Thomas, Brown, Richard J. P
• Format: Journal Article
• Language: English
• Published: 1752 N St., N.W., Washington, DC American Society for Microbiology 26.04.2021
• Subjects: Cellular Response to Infection; IFN regulatory factor 1 (IRF1); translational shutoff; hepatitis C virus (HCV); RNA-seq; primary human hepatocytes; eIF2 signaling; IFN signaling
• ISSN: 0022-538X; 1098-5514
• DOI: 10.1128/JVI.00245-21

Transcriptional profiling provides global snapshots of virus-mediated cellular reprogramming, which can simultaneously encompass pro- and antiviral components. To determine early transcriptional signatures associated with hepatitis C virus (HCV) infection of authentic target cells, we performed ex vivo infections of adult primary human hepatocytes (PHHs) from seven donors. Longitudinal sampling identified minimal gene dysregulation at 6 h postinfection (hpi). In contrast, at 72 hpi, massive increases in the breadth and magnitude of HCV-induced gene dysregulation were apparent, affecting gene classes associated with diverse biological processes. Comparison with HCV-induced transcriptional dysregulation in Huh-7.5 cells identified limited overlap between the two systems. Of note, in PHHs, HCV infection initiated broad upregulation of canonical interferon (IFN)-mediated defense programs, limiting viral RNA replication and abrogating virion release. We further find that the constitutive expression of IRF1 in PHHs maintains a steady-state antiviral program in the absence of infection, which can additionally reduce HCV RNA translation and replication. We also detected infection-induced downregulation of ∼90 genes encoding components of the eIF2 translation initiation complex and ribosomal subunits in PHHs, consistent with a signature of translational shutoff. As HCV polyprotein translation occurs independently of the eIF2 complex, this process is likely proviral: only translation initiation of host transcripts is arrested. The combination of antiviral intrinsic and inducible immunity, balanced against proviral programs, including translational arrest, maintains HCV replication at a low level in PHHs. This may ultimately keep HCV under the radar of extrahepatocyte immune surveillance while initial infection is established, promoting tolerance, preventing clearance, and facilitating progression to chronicity. IMPORTANCE Acute HCV infections are often asymptomatic and therefore frequently undiagnosed. We endeavored to recreate this understudied phase of HCV infection using explanted PHHs and monitored host responses to initial infection. We detected temporally distinct virus-induced perturbations in the transcriptional landscape, which were initially narrow but massively amplified in breadth and magnitude over time. At 72 hpi, we detected dysregulation of diverse gene programs, concurrently promoting both virus clearance and virus persistence. On the one hand, baseline expression of IRF1 combined with infection-induced upregulation of IFN-mediated effector genes suppresses virus propagation. On the other, we detect transcriptional signatures of host translational inhibition, which likely reduces the processing of IFN-regulated gene transcripts and facilitates virus survival. Together, our data provide important insights into constitutive and virus-induced transcriptional programs in PHHs and identify simultaneous antagonistic dysregulation of pro- and antiviral programs, which may facilitate host tolerance and promote viral persistence.