Journal article
Hepatitis C virus variants resistant to macrocyclic NS3-4A inhibitors subvert IFN-β induction by efficient MAVS cleavage.
- Welsch C Department of Internal Medicine 1, Goethe University Hospital Frankfurt, D-60590 Frankfurt a.M., Germany; Max-Planck-Institute for Informatics, Computational Biology & Applied Algorithmics, D-66123 Saarbrücken, Germany.
- Haselow K Department of Internal Medicine 1, Goethe University Hospital Frankfurt, D-60590 Frankfurt a.M., Germany.
- Gouttenoire J Division of Gastroenterology and Hepatology, Centre Hospitalier Universitaire Vaudois, University of Lausanne, CH-1011 Lausanne, Switzerland.
- Schneider M Technical University Munich, Center for Integrated Protein Science Munich (CIPS(M)) and Department of Life Science, D-85354 Freising-Weihenstephan, Germany.
- Morikawa K Division of Gastroenterology and Hepatology, Centre Hospitalier Universitaire Vaudois, University of Lausanne, CH-1011 Lausanne, Switzerland.
- Martinez Y Department of Internal Medicine 1, Goethe University Hospital Frankfurt, D-60590 Frankfurt a.M., Germany.
- Susser S Department of Internal Medicine 1, Goethe University Hospital Frankfurt, D-60590 Frankfurt a.M., Germany.
- Sarrazin C Department of Internal Medicine 1, Goethe University Hospital Frankfurt, D-60590 Frankfurt a.M., Germany.
- Zeuzem S Department of Internal Medicine 1, Goethe University Hospital Frankfurt, D-60590 Frankfurt a.M., Germany.
- Antes I Technical University Munich, Center for Integrated Protein Science Munich (CIPS(M)) and Department of Life Science, D-85354 Freising-Weihenstephan, Germany.
- Moradpour D Division of Gastroenterology and Hepatology, Centre Hospitalier Universitaire Vaudois, University of Lausanne, CH-1011 Lausanne, Switzerland.
- Lange CM Department of Internal Medicine 1, Goethe University Hospital Frankfurt, D-60590 Frankfurt a.M., Germany. Electronic address: Christian.Lange@kgu.de.
- 2014-12-03
Published in:
- Journal of hepatology. - 2015
All-oral therapy
Cardif
Direct-acting antiviral agent
Innate immunity
RIG-I signaling
Adaptor Proteins, Signal Transducing
Antiviral Agents
Cell Culture Techniques
DEAD Box Protein 58
DEAD-box RNA Helicases
Drug Resistance, Viral
Genotype
Hepacivirus
Hepatitis C, Chronic
Humans
Interferon-beta
RNA Helicases
RNA, Viral
Serine Endopeptidases
Viral Nonstructural Proteins
English
BACKGROUND & AIMS
The hepatitis C virus (HCV) NS3-4A protease is essential for the HCV life cycle and a prime target of antiviral treatment strategies. Protease inhibitors, however, are limited by emergence of resistance-associated amino acid variants (RAVs). The capacity to cleave and inactivate mitochondrial antiviral-signaling protein (MAVS) in the RIG-I-signaling pathway is a cardinal feature of NS3-4A, by which HCV blocks induction of interferon-(IFN)-β, thereby promoting viral persistence. Here, we aimed to investigate the impact of NS3-4A RAVs on MAVS cleavage.
METHODS
The impact of NS3-4A RAVs on MAVS cleavage was assessed using immunoblot analyses, luciferase reporter assays and molecular dynamics simulations to study the underlying molecular principles. IFN-β was quantified in serum from patients with different NS3-4A RAVs.
RESULTS
We show that macrocyclic NS3-4A RAVS with substitutions at residue D168 of the protease result in an increased capacity of NS3-4A to cleave MAVS and suppress IFN-β induction compared with a comprehensive panel of RAVs and wild type HCV. Mechanistically, we show the reconstitution of a tight network of electrostatic interactions between protease and the peptide substrate that allows much stronger binding of MAVS to D168 RAVs than to the wild-type protease. Accordingly, we could show IFN-β serum levels to be lower in patients with treatment failure due to the selection of D168 variants compared to R155 RAVs.
CONCLUSIONS
Our data constitutes a proof of concept that the selection of RAVs against specific classes of direct antivirals can lead to the predominance of viral variants with possibly adverse pathogenic characteristics.
The hepatitis C virus (HCV) NS3-4A protease is essential for the HCV life cycle and a prime target of antiviral treatment strategies. Protease inhibitors, however, are limited by emergence of resistance-associated amino acid variants (RAVs). The capacity to cleave and inactivate mitochondrial antiviral-signaling protein (MAVS) in the RIG-I-signaling pathway is a cardinal feature of NS3-4A, by which HCV blocks induction of interferon-(IFN)-β, thereby promoting viral persistence. Here, we aimed to investigate the impact of NS3-4A RAVs on MAVS cleavage.
METHODS
The impact of NS3-4A RAVs on MAVS cleavage was assessed using immunoblot analyses, luciferase reporter assays and molecular dynamics simulations to study the underlying molecular principles. IFN-β was quantified in serum from patients with different NS3-4A RAVs.
RESULTS
We show that macrocyclic NS3-4A RAVS with substitutions at residue D168 of the protease result in an increased capacity of NS3-4A to cleave MAVS and suppress IFN-β induction compared with a comprehensive panel of RAVs and wild type HCV. Mechanistically, we show the reconstitution of a tight network of electrostatic interactions between protease and the peptide substrate that allows much stronger binding of MAVS to D168 RAVs than to the wild-type protease. Accordingly, we could show IFN-β serum levels to be lower in patients with treatment failure due to the selection of D168 variants compared to R155 RAVs.
CONCLUSIONS
Our data constitutes a proof of concept that the selection of RAVs against specific classes of direct antivirals can lead to the predominance of viral variants with possibly adverse pathogenic characteristics.
- Language
-
- English
- Open access status
- closed
- Identifiers
-
- DOI 10.1016/j.jhep.2014.11.009
- PMID 25463536
- Persistent URL
- https://sonar.ch/global/documents/930
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