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

作者：Maria L. Agostini, Erica L. Andres, Amy Sims, Rachel L. Graham, Timothy P. Sheahan, Xiaotao Lu, Everett Clinton Smith, James Brett Case, Joy Y. Feng, Robert Jordan, Adrian S. Ray, Tomáš Cihlář, Dustin S. Siegel, Richard L. Mackman, Michael O. Clarke, Ralph S. Baric, Mark R. Denison · 发表于：mBio · 年份：2018 · DOI：10.1128/mbio.00221-18 · 被引用次数：1519 · 研究领域：SARS-CoV-2 and COVID-19 Research、Animal Virus Infections Studies、Viral gastroenteritis research and epidemiology

ABSTRACT 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 e...