Article Text

Original research
Efficient acute and chronic infection of stem cell-derived hepatocytes by hepatitis C virus
  1. Arnaud Carpentier1,2,
  2. Julie Sheldon1,
  3. Florian W R Vondran3,4,
  4. Richard JP Brown1,5,
  5. Thomas Pietschmann1,2,4
  1. 1 Institute of Experimental Virology, Twincore, Hannover, Germany
  2. 2 Cluster of Excellence RESIST (EXC 2155), Hannover Medical School, Hannover, Germany
  3. 3 ReMediES, Department of General, Visceral and Transplant Surgery, Hannover Medical School, Hannover, Germany
  4. 4 German Centre for Infection Research (DZIF), partner site Hannover-Braunschweig, Germany
  5. 5 Division of Veterinary Medicine, Paul-Ehrlich-Institut, Langen, Germany
  1. Correspondence to Dr Arnaud Carpentier, Twincore, Hannover, Germany; arnaud.carpentier{at}twincore.de; Professor Thomas Pietschmann; thomas.pietschmann{at}twincore.de

Abstract

Objective and design Human stem cell-derived hepatocyte-like cells (HLCs) have shown high potential as authentic model for dissection of the HCV life cycle and virus-induced pathogenesis. However, modest HCV replication, possibly due to robust innate immune responses, limits their broader use. To overcome these limitations and to dissect the mechanisms responsible for control of HCV, we analysed expression of key components of the interferon (IFN) system in HLCs, assessed permissiveness for different HCV strains and blocked innate immune signalling by pharmacological intervention.

Results Transcriptional profiling revealed that HLCs constitutively express messenger RNA of RLRs, and members of the IFN pathway. Moreover, HLCs upregulated IFNs and canonical interferon-regulated genes (IRGs) upon transfection with the double-stranded RNA mimic poly(I:C). Infection of HLCs with Jc1-HCVcc produced only limited viral progeny. In contrast, infection with p100, a Jc1-derived virus population with enhanced replication fitness and partial resistance to IFN, resulted in robust yet transient viraemia. Viral titres declined concomitant with a peak of IRG induction. Addition of ruxolitinib, a JAK/STAT inhibitor, permitted chronic infection and raised p100 infectious virus titres to 1×105 FFU/mL. IRGs expression profiling in infected HLCs revealed a landscape of HCV-dependent transcriptional changes similar to HCV-infected primary human hepatocytes, but distinct from Huh-7.5 cells. Withdrawal of ruxolitinib restored innate immune responses and resulted in HCV clearance.

Conclusion This authentic human cell model is well suited to examine acute and chronic host-HCV interactions, particularly IFN-triggered antiviral effector functions and mechanisms of innate immune control of HCV infection.

  • stem cells
  • hepatocyte
  • HCV
  • immune response
  • interferon
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Footnotes

  • Contributors AC and TP designed the study concept, analysed data and wrote the manuscript. AC also performed experiments. JS and RB performed experiments, analysed data and edited the manuscript. FWRV provided essential reagents and edited the manuscript.

  • Funding This study is funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation)—Project ID 158989968—SFB900, subprojects A6 and under the Germany’s Excellence Strategy—EXC 2155 'RESIST'—Project ID 39087428.

  • Disclaimer The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript.

  • Competing interests None declared.

  • Patient and public involvement Patients and/or the public were not involved in the design, conduct, reporting or dissemination plans of this research.

  • Patient consent for publication Not required.

  • Provenance and peer review Not commissioned; externally peer reviewed.

  • Data availability statement Data are available upon reasonable request to: Dr Arnaud Carpentier, arnaud.carpentier@twincore.de, ORCID identifier 0000-0002-6994-4689; Professor Thomas Pietschmann, thomas.pietschmann@twincore.de, ORCID identifier 0000-0001-6789-4422.

    RNA-seq data are publicly available through the NIH GEO platform (https://www.ncbi.nlm.nih.gov/geo/): GEO accession GSE132606 and GSE132548.

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