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Identification of the Niemann-Pick C1–like 1 cholesterol absorption receptor as a new hepatitis C virus entry factor

Abstract

Hepatitis C virus (HCV) is a leading cause of liver disease worldwide. With 170 million individuals infected and current interferon-based treatment having toxic side effects and marginal efficacy, more effective antivirals are crucially needed1. Although HCV protease inhibitors were just approved by the US Food and Drug Administration (FDA), optimal HCV therapy, analogous to HIV therapy, will probably require a combination of antivirals targeting multiple aspects of the viral lifecycle. Viral entry represents a potential multifaceted target for antiviral intervention; however, to date, FDA-approved inhibitors of HCV cell entry are unavailable. Here we show that the cellular Niemann-Pick C1–like 1 (NPC1L1) cholesterol uptake receptor is an HCV entry factor amendable to therapeutic intervention. Specifically, NPC1L1 expression is necessary for HCV infection, as silencing or antibody-mediated blocking of NPC1L1 impairs cell culture–derived HCV (HCVcc) infection initiation. In addition, the clinically available FDA-approved NPC1L1 antagonist ezetimibe2,3 potently blocks HCV uptake in vitro via a virion cholesterol–dependent step before virion-cell membrane fusion. Moreover, ezetimibe inhibits infection by all major HCV genotypes in vitro and in vivo delays the establishment of HCV genotype 1b infection in mice with human liver grafts. Thus, we have not only identified NPC1L1 as an HCV cell entry factor but also discovered a new antiviral target and potential therapeutic agent.

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Figure 1: NPC1L1 has a role in HCVcc infection.
Figure 2: Ezetimibe-mediated inhibition of NPC1L1 reduces HCV entry at a post-binding, prefusion step.
Figure 3: NPC1L1-mediated HCV cell entry is cholesterol dependent.
Figure 4: Ezetimibe delays the establishment of HCV infection in hepatic xenorepopulated mice.

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Acknowledgements

We thank T. Wakita (National Institute of Infectious Diseases, Japan) for JFH1-based plasmids, F. Chisari (The Scripps Research Institute) for Huh7 cells, J. Buhk (Copenhagen University Hospital Hepatitis C Program) for JFH-1–based intergenotypic HCV clones, Y. Ioannou (Mount Sinai School of Medicine) for the antibody (Bsn4052) against NPC1L1, D. Burton and M. Law (The Scripps Research Institute) for the antibody (C1) against the HCV glycoprotein E2 and C. Rice (The Rockefeller Institute) for the antibody (E910) against the HCV nonstructural protein NS5A. We would also like to thank P. Corcoran for outstanding technical assistance, H. Dahari for assistance with statistical analyses and T. Layden and S. Cotler for editing the manuscript. We also thank J. Graves of the University of Illinois at Chicago (UIC) Research Resources Center Flow Cytometry laboratory and K. Ma of the UIC Research Resources Center Confocal Microscopy laboratory for technical assistance. This work was supported by the US National Institutes of Health Public Health Service grants R01-AI070827 and R03-AI085226 (S.L.U.), the American Cancer Society Research Scholar Grant RSG-09-076-01 (S.L.U.), the UIC Center for Clinical and Translational Science NIH grant UL1RR029879, the UIC Council to Support Gastrointestinal and Liver Disease and a grant from the Ministry of Education, Culture, Sports, Science and Technology-Japan, Ministry of Health, Labour and Welfare-Japan (K.C.).

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Contributions

B.S. made the initial discovery. B.S. and S.L.U. designed the project, analyzed the results and wrote the manuscript. B.S., N.B., D.N.M., S.H., K.A.M. and X.Y. performed experimental work. B.S., S.L.U., M.I. and K.C. designed the hepatic xenorepopulation mouse experiments, and N.H. performed the in vivo studies. W.A.A. was involved in the initial conception of the project and provided valuable expertise.

Corresponding authors

Correspondence to Bruno Sainz Jr or Susan L Uprichard.

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Competing interests

B.S. and S.L.U. declare a competing financial interest in the form of a patent application to the US Patent Office entitled "Methods for treatment of hepatitis C" (Appl. No. 61/093,549; filed September 2, 2008 and Appl. No. 61/169,899; filed April 16, 2009), which, in part, is based on and related to the findings described within the manuscript.

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Sainz, B., Barretto, N., Martin, D. et al. Identification of the Niemann-Pick C1–like 1 cholesterol absorption receptor as a new hepatitis C virus entry factor. Nat Med 18, 281–285 (2012). https://doi.org/10.1038/nm.2581

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