A comprehensive study of SARS-CoV-2 main protease (Mpro) inhibitor-resistant mutants selected in a VSV-based system.

Nirmatrelvir was the first protease inhibitor specifically developed against the SARS-CoV-2 main protease (3CLpro/Mpro) and licensed for clinical use. As SARS-CoV-2 continues to spread, variants resistant to nirmatrelvir and other currently available treatments are likely to arise. This study aimed...

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Main Authors: Francesco Costacurta, Andrea Dodaro, David Bante, Helge Schöppe, Ju-Yi Peng, Bernhard Sprenger, Xi He, Seyed Arad Moghadasi, Lisa Maria Egger, Jakob Fleischmann, Matteo Pavan, Davide Bassani, Silvia Menin, Stefanie Rauch, Laura Krismer, Anna Sauerwein, Anne Heberle, Toni Rabensteiner, Joses Ho, Reuben S Harris, Eduard Stefan, Rainer Schneider, Theresia Dunzendorfer-Matt, Andreas Naschberger, Dai Wang, Teresa Kaserer, Stefano Moro, Dorothee von Laer, Emmanuel Heilmann
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2024-09-01
Series:PLoS Pathogens
Online Access:https://doi.org/10.1371/journal.ppat.1012522
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author Francesco Costacurta
Andrea Dodaro
David Bante
Helge Schöppe
Ju-Yi Peng
Bernhard Sprenger
Xi He
Seyed Arad Moghadasi
Lisa Maria Egger
Jakob Fleischmann
Matteo Pavan
Davide Bassani
Silvia Menin
Stefanie Rauch
Laura Krismer
Anna Sauerwein
Anne Heberle
Toni Rabensteiner
Joses Ho
Reuben S Harris
Eduard Stefan
Rainer Schneider
Theresia Dunzendorfer-Matt
Andreas Naschberger
Dai Wang
Teresa Kaserer
Stefano Moro
Dorothee von Laer
Emmanuel Heilmann
author_facet Francesco Costacurta
Andrea Dodaro
David Bante
Helge Schöppe
Ju-Yi Peng
Bernhard Sprenger
Xi He
Seyed Arad Moghadasi
Lisa Maria Egger
Jakob Fleischmann
Matteo Pavan
Davide Bassani
Silvia Menin
Stefanie Rauch
Laura Krismer
Anna Sauerwein
Anne Heberle
Toni Rabensteiner
Joses Ho
Reuben S Harris
Eduard Stefan
Rainer Schneider
Theresia Dunzendorfer-Matt
Andreas Naschberger
Dai Wang
Teresa Kaserer
Stefano Moro
Dorothee von Laer
Emmanuel Heilmann
author_sort Francesco Costacurta
collection DOAJ
description Nirmatrelvir was the first protease inhibitor specifically developed against the SARS-CoV-2 main protease (3CLpro/Mpro) and licensed for clinical use. As SARS-CoV-2 continues to spread, variants resistant to nirmatrelvir and other currently available treatments are likely to arise. This study aimed to identify and characterize mutations that confer resistance to nirmatrelvir. To safely generate Mpro resistance mutations, we passaged a previously developed, chimeric vesicular stomatitis virus (VSV-Mpro) with increasing, yet suboptimal concentrations of nirmatrelvir. Using Wuhan-1 and Omicron Mpro variants, we selected a large set of mutants. Some mutations are frequently present in GISAID, suggesting their relevance in SARS-CoV-2. The resistance phenotype of a subset of mutations was characterized against clinically available protease inhibitors (nirmatrelvir and ensitrelvir) with cell-based, biochemical and SARS-CoV-2 replicon assays. Moreover, we showed the putative molecular mechanism of resistance based on in silico molecular modelling. These findings have implications on the development of future generation Mpro inhibitors, will help to understand SARS-CoV-2 protease inhibitor resistance mechanisms and show the relevance of specific mutations, thereby informing treatment decisions.
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spelling doaj-art-8a2d120a7782489fb1d69e04556b84182025-08-20T02:14:43ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742024-09-01209e101252210.1371/journal.ppat.1012522A comprehensive study of SARS-CoV-2 main protease (Mpro) inhibitor-resistant mutants selected in a VSV-based system.Francesco CostacurtaAndrea DodaroDavid BanteHelge SchöppeJu-Yi PengBernhard SprengerXi HeSeyed Arad MoghadasiLisa Maria EggerJakob FleischmannMatteo PavanDavide BassaniSilvia MeninStefanie RauchLaura KrismerAnna SauerweinAnne HeberleToni RabensteinerJoses HoReuben S HarrisEduard StefanRainer SchneiderTheresia Dunzendorfer-MattAndreas NaschbergerDai WangTeresa KasererStefano MoroDorothee von LaerEmmanuel HeilmannNirmatrelvir was the first protease inhibitor specifically developed against the SARS-CoV-2 main protease (3CLpro/Mpro) and licensed for clinical use. As SARS-CoV-2 continues to spread, variants resistant to nirmatrelvir and other currently available treatments are likely to arise. This study aimed to identify and characterize mutations that confer resistance to nirmatrelvir. To safely generate Mpro resistance mutations, we passaged a previously developed, chimeric vesicular stomatitis virus (VSV-Mpro) with increasing, yet suboptimal concentrations of nirmatrelvir. Using Wuhan-1 and Omicron Mpro variants, we selected a large set of mutants. Some mutations are frequently present in GISAID, suggesting their relevance in SARS-CoV-2. The resistance phenotype of a subset of mutations was characterized against clinically available protease inhibitors (nirmatrelvir and ensitrelvir) with cell-based, biochemical and SARS-CoV-2 replicon assays. Moreover, we showed the putative molecular mechanism of resistance based on in silico molecular modelling. These findings have implications on the development of future generation Mpro inhibitors, will help to understand SARS-CoV-2 protease inhibitor resistance mechanisms and show the relevance of specific mutations, thereby informing treatment decisions.https://doi.org/10.1371/journal.ppat.1012522
spellingShingle Francesco Costacurta
Andrea Dodaro
David Bante
Helge Schöppe
Ju-Yi Peng
Bernhard Sprenger
Xi He
Seyed Arad Moghadasi
Lisa Maria Egger
Jakob Fleischmann
Matteo Pavan
Davide Bassani
Silvia Menin
Stefanie Rauch
Laura Krismer
Anna Sauerwein
Anne Heberle
Toni Rabensteiner
Joses Ho
Reuben S Harris
Eduard Stefan
Rainer Schneider
Theresia Dunzendorfer-Matt
Andreas Naschberger
Dai Wang
Teresa Kaserer
Stefano Moro
Dorothee von Laer
Emmanuel Heilmann
A comprehensive study of SARS-CoV-2 main protease (Mpro) inhibitor-resistant mutants selected in a VSV-based system.
PLoS Pathogens
title A comprehensive study of SARS-CoV-2 main protease (Mpro) inhibitor-resistant mutants selected in a VSV-based system.
title_full A comprehensive study of SARS-CoV-2 main protease (Mpro) inhibitor-resistant mutants selected in a VSV-based system.
title_fullStr A comprehensive study of SARS-CoV-2 main protease (Mpro) inhibitor-resistant mutants selected in a VSV-based system.
title_full_unstemmed A comprehensive study of SARS-CoV-2 main protease (Mpro) inhibitor-resistant mutants selected in a VSV-based system.
title_short A comprehensive study of SARS-CoV-2 main protease (Mpro) inhibitor-resistant mutants selected in a VSV-based system.
title_sort comprehensive study of sars cov 2 main protease mpro inhibitor resistant mutants selected in a vsv based system
url https://doi.org/10.1371/journal.ppat.1012522
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