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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)...
Autores principales: | , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Microbiology
2018
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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 |
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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 |
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