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EGFR and EphA2 are host factors for hepatitis C virus entry and possible targets for antiviral therapy

Abstract

Hepatitis C virus (HCV) is a major cause of liver disease, but therapeutic options are limited and there are no prevention strategies. Viral entry is the first step of infection and requires the cooperative interaction of several host cell factors. Using a functional RNAi kinase screen, we identified epidermal growth factor receptor and ephrin receptor A2 as host cofactors for HCV entry. Blocking receptor kinase activity by approved inhibitors broadly impaired infection by all major HCV genotypes and viral escape variants in cell culture and in a human liver chimeric mouse model in vivo. The identified receptor tyrosine kinases (RTKs) mediate HCV entry by regulating CD81–claudin-1 co-receptor associations and viral glycoprotein–dependent membrane fusion. These results identify RTKs as previously unknown HCV entry cofactors and show that tyrosine kinase inhibitors have substantial antiviral activity. Inhibition of RTK function may constitute a new approach for prevention and treatment of HCV infection.

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Figure 1: EGFR is a cofactor for HCV entry.
Figure 2: Inhibition of EGFR activation by kinase inhibitors reduces HCV entry and infection.
Figure 3: Modulation of HCV entry by EGFR ligands and an EGFR-specific antibody.
Figure 4: EGFR mediates HCV entry at postbinding steps by promoting CD81-CLDN1 co-receptor interactions and membrane fusion.
Figure 5: Functional role of EGFR in viral cell-to-cell transmission and spread.
Figure 6: Erlotinib modulates HCV kinetics and inhibits infection in vivo.

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Acknowledgements

This work was supported by the European Research Council (ERC-2008-AdG-233130-HEPCENT), INTERREG-IV-Rhin Supérieur-FEDER-Hepato-Regio-Net 2009, Agence Nationale de la Recherche (ANR-05-CEXC-008), Agence Nationale de Recherche sur le Sida 2008/354, Région Alsace, Institut National du Cancer, the Institut de Génétique et de Biologie Moléculaire et Cellulaire, Institut National de la Santé et de la Recherche Médicale, Centre National de la Recherche Scientifique, Université de Strasbourg, the US National Insititutes of Health (1K08DE020139-01A1), the UK Medical Research Council and the Wellcome Trust. We acknowledge A.-L. Morand, L. Froidevaux, A. Weiss, L. Poidevin, S. Durand and E. Soulier for excellent technical work. We thank R. Bartenschlager (University of Heidelberg) for providing Jc1 and Luc-Jc1 expression vectors, J. Ball (University of Nottingham) for UKN2A.2.4, UKN3A1.28 and UKN4.21.16 expression vectors, T. Wakita (National Institute of Infectious Diseases, Japan) for JFH1 constructs, C.M. Rice (The Rockefeller University) for Huh7.5 cells, F.V. Chisari (The Scripps Research Institute) for Huh7.5.1 cells, E. Harlow (Harvard University) for CDC2 expression plasmids and M. Tanaka (Hamamatsu University) for EphA2 expression plasmids.

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J.L., M.B.Z. and T.F.B. wrote the manuscript. J.L., M.B.Z., F.X., D.L., F.-L.C., J.A.M., and T.F.B. designed experiments and analyzed data. J.L., M.B.Z., F.X., C.T., I.F., L.Z., C.D., C.J.M., M.T., S.G., C.R., M.N.Z., D.L. and J.F. performed experiments. S.M.R., T.P., A.H.P., P.P. and M.D. contributed essential reagents. W.R. and O.P. performed bioinformatic analyses. J.L., B.F. and L.B. implemented and coordinated the siRNA screen. T.F.B. designed and supervised the project.

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Correspondence to Thomas F Baumert.

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Lupberger, J., Zeisel, M., Xiao, F. et al. EGFR and EphA2 are host factors for hepatitis C virus entry and possible targets for antiviral therapy. Nat Med 17, 589–595 (2011). https://doi.org/10.1038/nm.2341

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