The A150V Polymorphism of Genotype 3 Hepatitis C Virus Polymerase Inhibits Interferon Alfa by Suppressing Protein Kinase R Activation
| Autor: | Swathi Rajagopal, Graham R. Foster, Meleri Jones, Peter A C Wing, Wing-Yiu Jason Lee |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
viruses Protein Kinase R Apoptosis G3 genotype 3 Hepacivirus Viral Nonstructural Proteins Virus Replication medicine.disease_cause eIF-2 Kinase chemistry.chemical_compound 0302 clinical medicine Interferon Tumor Cells Cultured Polymerase Original Research biology Liver Neoplasms Gastroenterology Hepatitis C 17-AAG geldanamycin analogue Real-time polymerase chain reaction HCV hepatitis C virus RNA Viral 030211 gastroenterology & hepatology medicine.drug Hepatitis C Virus FSC forward scatter Carcinoma Hepatocellular Hepatitis C virus PBS phosphate-buffered saline Alpha interferon SEM standard error of the mean Antiviral Agents PKR protein kinase R 03 medical and health sciences 2-AP 2-aminopurine medicine ISG interferon-stimulated gene Humans IFN interferon lcsh:RC799-869 NS5B Interferon alfa Polymorphism Genetic Hepatology ISGF3 interferon-stimulated factor 3 RT-qPCR reverse transcriptase quantitative polymerase chain reaction Interferon-alpha DMEM Dulbecco modified Eagle medium PE phycoerythrin RNA-Dependent RNA Polymerase WT wild-type Virology Protein kinase R Viral Replication 030104 developmental biology chemistry biology.protein dsRNA double-stranded RNA lcsh:Diseases of the digestive system. Gastroenterology Replicon |
| Zdroj: | Cellular and Molecular Gastroenterology and Hepatology Cellular and Molecular Gastroenterology and Hepatology, Vol 11, Iss 4, Pp 1163-1175 (2021) |
| ISSN: | 2352-345X |
| DOI: | 10.1016/j.jcmgh.2020.11.012 |
| Popis: | Background & Aims Despite recent advances in antiviral therapy for hepatitis C virus (HCV), a proportion of patients with genotype 3 (G3) HCV infection do not respond to current all oral treatment regimens. Genomic analyses have identified key polymorphisms correlating with increased resistance to direct-acting antivirals. We previously reported that amino acid polymorphisms (A150V and K206E) in the polymerase (NS5B) of G3 HCV reduce response to sofosbuvir. We now demonstrate that these polymorphisms alter the response to interferon alpha. Methods Quantitative polymerase chain reaction, immunofluorescence, luciferase activity assay, immunoblotting, and flow cytometry were used to study the antiviral effect of interferon (IFN) on DBN G3 HCV-infected cells and G3 HCV replicons. Results We show the presence of the A150V polymorphism markedly reduces the response to IFN alpha (IC50 of S52_WT = 1.162 IU/mL and IC50 of S52_A150V = 14.45 IU/mL, 12.4-fold difference). The induction of IFN-stimulated genes in A150V replicon cells is unaffected, but nuclear localization of active protein kinase R (PKR) is reduced. Blockade of PKR activity reduced the antiviral effect of IFN on wild-type replicons, whereas augmented PKR activation promoted the antiviral effect of IFN on A150V replicons. Furthermore, we show that impaired activation of PKR in A150V replicon cells diminishes cellular apoptosis. Conclusions These results demonstrate that polymorphisms reducing response rates to direct-acting antivirals may function beyond conferring drug resistance by modulating the intrinsic cellular antiviral response. |
| Databáze: | OpenAIRE |
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