Cargando…

Coronavirus Susceptibility to the Antiviral Remdesivir (GS-5734) Is Mediated by the Viral Polymerase and the Proofreading Exoribonuclease

Emerging coronaviruses (CoVs) cause severe disease in humans, but no approved therapeutics are available. The CoV nsp14 exoribonuclease (ExoN) has complicated development of antiviral nucleosides due to its proofreading activity. We recently reported that the nucleoside analogue GS-5734 (remdesivir)...

Descripción completa

Detalles Bibliográficos
Autores principales: Agostini, Maria L., Andres, Erica L., Sims, Amy C., Graham, Rachel L., Sheahan, Timothy P., Lu, Xiaotao, Smith, Everett Clinton, Case, James Brett, Feng, Joy Y., Jordan, Robert, Ray, Adrian S., Cihlar, Tomas, Siegel, Dustin, Mackman, Richard L., Clarke, Michael O., Baric, Ralph S., Denison, Mark R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Microbiology 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5844999/
https://www.ncbi.nlm.nih.gov/pubmed/29511076
http://dx.doi.org/10.1128/mBio.00221-18
_version_ 1783305337120489472
author Agostini, Maria L.
Andres, Erica L.
Sims, Amy C.
Graham, Rachel L.
Sheahan, Timothy P.
Lu, Xiaotao
Smith, Everett Clinton
Case, James Brett
Feng, Joy Y.
Jordan, Robert
Ray, Adrian S.
Cihlar, Tomas
Siegel, Dustin
Mackman, Richard L.
Clarke, Michael O.
Baric, Ralph S.
Denison, Mark R.
author_facet Agostini, Maria L.
Andres, Erica L.
Sims, Amy C.
Graham, Rachel L.
Sheahan, Timothy P.
Lu, Xiaotao
Smith, Everett Clinton
Case, James Brett
Feng, Joy Y.
Jordan, Robert
Ray, Adrian S.
Cihlar, Tomas
Siegel, Dustin
Mackman, Richard L.
Clarke, Michael O.
Baric, Ralph S.
Denison, Mark R.
author_sort Agostini, Maria L.
collection PubMed
description Emerging coronaviruses (CoVs) cause severe disease in humans, but no approved therapeutics are available. The CoV nsp14 exoribonuclease (ExoN) has complicated development of antiviral nucleosides due to its proofreading activity. We recently reported that the nucleoside analogue GS-5734 (remdesivir) potently inhibits human and zoonotic CoVs in vitro and in a severe acute respiratory syndrome coronavirus (SARS-CoV) mouse model. However, studies with GS-5734 have not reported resistance associated with GS-5734, nor do we understand the action of GS-5734 in wild-type (WT) proofreading CoVs. Here, we show that GS-5734 inhibits murine hepatitis virus (MHV) with similar 50% effective concentration values (EC(50)) as SARS-CoV and Middle East respiratory syndrome coronavirus (MERS-CoV). Passage of WT MHV in the presence of the GS-5734 parent nucleoside selected two mutations in the nsp12 polymerase at residues conserved across all CoVs that conferred up to 5.6-fold resistance to GS-5734, as determined by EC(50). The resistant viruses were unable to compete with WT in direct coinfection passage in the absence of GS-5734. Introduction of the MHV resistance mutations into SARS-CoV resulted in the same in vitro resistance phenotype and attenuated SARS-CoV pathogenesis in a mouse model. Finally, we demonstrate that an MHV mutant lacking ExoN proofreading was significantly more sensitive to GS-5734. Combined, the results indicate that GS-5734 interferes with the nsp12 polymerase even in the setting of intact ExoN proofreading activity and that resistance can be overcome with increased, nontoxic concentrations of GS-5734, further supporting the development of GS-5734 as a broad-spectrum therapeutic to protect against contemporary and emerging CoVs.
format Online
Article
Text
id pubmed-5844999
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher American Society for Microbiology
record_format MEDLINE/PubMed
spelling pubmed-58449992018-03-21 Coronavirus Susceptibility to the Antiviral Remdesivir (GS-5734) Is Mediated by the Viral Polymerase and the Proofreading Exoribonuclease Agostini, Maria L. Andres, Erica L. Sims, Amy C. Graham, Rachel L. Sheahan, Timothy P. Lu, Xiaotao Smith, Everett Clinton Case, James Brett Feng, Joy Y. Jordan, Robert Ray, Adrian S. Cihlar, Tomas Siegel, Dustin Mackman, Richard L. Clarke, Michael O. Baric, Ralph S. Denison, Mark R. mBio Research Article Emerging coronaviruses (CoVs) cause severe disease in humans, but no approved therapeutics are available. The CoV nsp14 exoribonuclease (ExoN) has complicated development of antiviral nucleosides due to its proofreading activity. We recently reported that the nucleoside analogue GS-5734 (remdesivir) potently inhibits human and zoonotic CoVs in vitro and in a severe acute respiratory syndrome coronavirus (SARS-CoV) mouse model. However, studies with GS-5734 have not reported resistance associated with GS-5734, nor do we understand the action of GS-5734 in wild-type (WT) proofreading CoVs. Here, we show that GS-5734 inhibits murine hepatitis virus (MHV) with similar 50% effective concentration values (EC(50)) as SARS-CoV and Middle East respiratory syndrome coronavirus (MERS-CoV). Passage of WT MHV in the presence of the GS-5734 parent nucleoside selected two mutations in the nsp12 polymerase at residues conserved across all CoVs that conferred up to 5.6-fold resistance to GS-5734, as determined by EC(50). The resistant viruses were unable to compete with WT in direct coinfection passage in the absence of GS-5734. Introduction of the MHV resistance mutations into SARS-CoV resulted in the same in vitro resistance phenotype and attenuated SARS-CoV pathogenesis in a mouse model. Finally, we demonstrate that an MHV mutant lacking ExoN proofreading was significantly more sensitive to GS-5734. Combined, the results indicate that GS-5734 interferes with the nsp12 polymerase even in the setting of intact ExoN proofreading activity and that resistance can be overcome with increased, nontoxic concentrations of GS-5734, further supporting the development of GS-5734 as a broad-spectrum therapeutic to protect against contemporary and emerging CoVs. American Society for Microbiology 2018-03-06 /pmc/articles/PMC5844999/ /pubmed/29511076 http://dx.doi.org/10.1128/mBio.00221-18 Text en Copyright © 2018 Agostini et al. https://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Research Article
Agostini, Maria L.
Andres, Erica L.
Sims, Amy C.
Graham, Rachel L.
Sheahan, Timothy P.
Lu, Xiaotao
Smith, Everett Clinton
Case, James Brett
Feng, Joy Y.
Jordan, Robert
Ray, Adrian S.
Cihlar, Tomas
Siegel, Dustin
Mackman, Richard L.
Clarke, Michael O.
Baric, Ralph S.
Denison, Mark R.
Coronavirus Susceptibility to the Antiviral Remdesivir (GS-5734) Is Mediated by the Viral Polymerase and the Proofreading Exoribonuclease
title Coronavirus Susceptibility to the Antiviral Remdesivir (GS-5734) Is Mediated by the Viral Polymerase and the Proofreading Exoribonuclease
title_full Coronavirus Susceptibility to the Antiviral Remdesivir (GS-5734) Is Mediated by the Viral Polymerase and the Proofreading Exoribonuclease
title_fullStr Coronavirus Susceptibility to the Antiviral Remdesivir (GS-5734) Is Mediated by the Viral Polymerase and the Proofreading Exoribonuclease
title_full_unstemmed Coronavirus Susceptibility to the Antiviral Remdesivir (GS-5734) Is Mediated by the Viral Polymerase and the Proofreading Exoribonuclease
title_short Coronavirus Susceptibility to the Antiviral Remdesivir (GS-5734) Is Mediated by the Viral Polymerase and the Proofreading Exoribonuclease
title_sort coronavirus susceptibility to the antiviral remdesivir (gs-5734) is mediated by the viral polymerase and the proofreading exoribonuclease
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5844999/
https://www.ncbi.nlm.nih.gov/pubmed/29511076
http://dx.doi.org/10.1128/mBio.00221-18
work_keys_str_mv AT agostinimarial coronavirussusceptibilitytotheantiviralremdesivirgs5734ismediatedbytheviralpolymeraseandtheproofreadingexoribonuclease
AT andreserical coronavirussusceptibilitytotheantiviralremdesivirgs5734ismediatedbytheviralpolymeraseandtheproofreadingexoribonuclease
AT simsamyc coronavirussusceptibilitytotheantiviralremdesivirgs5734ismediatedbytheviralpolymeraseandtheproofreadingexoribonuclease
AT grahamrachell coronavirussusceptibilitytotheantiviralremdesivirgs5734ismediatedbytheviralpolymeraseandtheproofreadingexoribonuclease
AT sheahantimothyp coronavirussusceptibilitytotheantiviralremdesivirgs5734ismediatedbytheviralpolymeraseandtheproofreadingexoribonuclease
AT luxiaotao coronavirussusceptibilitytotheantiviralremdesivirgs5734ismediatedbytheviralpolymeraseandtheproofreadingexoribonuclease
AT smitheverettclinton coronavirussusceptibilitytotheantiviralremdesivirgs5734ismediatedbytheviralpolymeraseandtheproofreadingexoribonuclease
AT casejamesbrett coronavirussusceptibilitytotheantiviralremdesivirgs5734ismediatedbytheviralpolymeraseandtheproofreadingexoribonuclease
AT fengjoyy coronavirussusceptibilitytotheantiviralremdesivirgs5734ismediatedbytheviralpolymeraseandtheproofreadingexoribonuclease
AT jordanrobert coronavirussusceptibilitytotheantiviralremdesivirgs5734ismediatedbytheviralpolymeraseandtheproofreadingexoribonuclease
AT rayadrians coronavirussusceptibilitytotheantiviralremdesivirgs5734ismediatedbytheviralpolymeraseandtheproofreadingexoribonuclease
AT cihlartomas coronavirussusceptibilitytotheantiviralremdesivirgs5734ismediatedbytheviralpolymeraseandtheproofreadingexoribonuclease
AT siegeldustin coronavirussusceptibilitytotheantiviralremdesivirgs5734ismediatedbytheviralpolymeraseandtheproofreadingexoribonuclease
AT mackmanrichardl coronavirussusceptibilitytotheantiviralremdesivirgs5734ismediatedbytheviralpolymeraseandtheproofreadingexoribonuclease
AT clarkemichaelo coronavirussusceptibilitytotheantiviralremdesivirgs5734ismediatedbytheviralpolymeraseandtheproofreadingexoribonuclease
AT baricralphs coronavirussusceptibilitytotheantiviralremdesivirgs5734ismediatedbytheviralpolymeraseandtheproofreadingexoribonuclease
AT denisonmarkr coronavirussusceptibilitytotheantiviralremdesivirgs5734ismediatedbytheviralpolymeraseandtheproofreadingexoribonuclease