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Influenza A virus polymerase acidic protein E23G/K substitutions weaken key baloxavir drug-binding contacts with minimal impact on replication and transmission

Baloxavir marboxil (BXM) is approved for treating uncomplicated influenza. The active metabolite baloxavir acid (BXA) inhibits cap-dependent endonuclease activity of the influenza virus polymerase acidic protein (PA), which is necessary for viral transcription. Treatment-emergent E23G or E23K (E23G/...

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Autores principales: Jones, Jeremy C., Zagribelnyy, Bogdan, Pascua, Philippe Noriel Q., Bezrukov, Dmitry S., Barman, Subrata, Okda, Faten, Webby, Richard J., Ivanenkov, Yan A., Govorkova, Elena A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9312377/
https://www.ncbi.nlm.nih.gov/pubmed/35830486
http://dx.doi.org/10.1371/journal.ppat.1010698
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author Jones, Jeremy C.
Zagribelnyy, Bogdan
Pascua, Philippe Noriel Q.
Bezrukov, Dmitry S.
Barman, Subrata
Okda, Faten
Webby, Richard J.
Ivanenkov, Yan A.
Govorkova, Elena A.
author_facet Jones, Jeremy C.
Zagribelnyy, Bogdan
Pascua, Philippe Noriel Q.
Bezrukov, Dmitry S.
Barman, Subrata
Okda, Faten
Webby, Richard J.
Ivanenkov, Yan A.
Govorkova, Elena A.
author_sort Jones, Jeremy C.
collection PubMed
description Baloxavir marboxil (BXM) is approved for treating uncomplicated influenza. The active metabolite baloxavir acid (BXA) inhibits cap-dependent endonuclease activity of the influenza virus polymerase acidic protein (PA), which is necessary for viral transcription. Treatment-emergent E23G or E23K (E23G/K) PA substitutions have been implicated in reduced BXA susceptibility, but their effect on virus fitness and transmissibility, their synergism with other BXA resistance markers, and the mechanisms of resistance have been insufficiently studied. Accordingly, we generated point mutants of circulating seasonal influenza A(H1N1)pdm09 and A(H3N2) viruses carrying E23G/K substitutions. Both substitutions caused 2- to 13-fold increases in the BXA EC(50). EC(50)s were higher with E23K than with E23G and increased dramatically (138- to 446-fold) when these substitutions were combined with PA I38T, the dominant BXA resistance marker. E23G/K-substituted viruses exhibited slightly impaired replication in MDCK and Calu-3 cells, which was more pronounced with E23K. In ferret transmission experiments, all viruses transmitted to direct-contact and airborne-transmission animals, with only E23K+I38T viruses failing to infect 100% of animals by airborne transmission. E23G/K genotypes were predominantly stable during transmission events and through five passages in vitro. Thermostable PA–BXA interactions were weakened by E23G/K substitutions and further weakened when combined with I38T. In silico modeling indicated this was caused by E23G/K altering the placement of functionally important Tyr24 in the endonuclease domain, potentially decreasing BXA binding but at some cost to the virus. These data implicate E23G/K, alone or combined with I38T, as important markers of reduced BXM susceptibility, and such mutants could emerge and/or transmit among humans.
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spelling pubmed-93123772022-07-26 Influenza A virus polymerase acidic protein E23G/K substitutions weaken key baloxavir drug-binding contacts with minimal impact on replication and transmission Jones, Jeremy C. Zagribelnyy, Bogdan Pascua, Philippe Noriel Q. Bezrukov, Dmitry S. Barman, Subrata Okda, Faten Webby, Richard J. Ivanenkov, Yan A. Govorkova, Elena A. PLoS Pathog Research Article Baloxavir marboxil (BXM) is approved for treating uncomplicated influenza. The active metabolite baloxavir acid (BXA) inhibits cap-dependent endonuclease activity of the influenza virus polymerase acidic protein (PA), which is necessary for viral transcription. Treatment-emergent E23G or E23K (E23G/K) PA substitutions have been implicated in reduced BXA susceptibility, but their effect on virus fitness and transmissibility, their synergism with other BXA resistance markers, and the mechanisms of resistance have been insufficiently studied. Accordingly, we generated point mutants of circulating seasonal influenza A(H1N1)pdm09 and A(H3N2) viruses carrying E23G/K substitutions. Both substitutions caused 2- to 13-fold increases in the BXA EC(50). EC(50)s were higher with E23K than with E23G and increased dramatically (138- to 446-fold) when these substitutions were combined with PA I38T, the dominant BXA resistance marker. E23G/K-substituted viruses exhibited slightly impaired replication in MDCK and Calu-3 cells, which was more pronounced with E23K. In ferret transmission experiments, all viruses transmitted to direct-contact and airborne-transmission animals, with only E23K+I38T viruses failing to infect 100% of animals by airborne transmission. E23G/K genotypes were predominantly stable during transmission events and through five passages in vitro. Thermostable PA–BXA interactions were weakened by E23G/K substitutions and further weakened when combined with I38T. In silico modeling indicated this was caused by E23G/K altering the placement of functionally important Tyr24 in the endonuclease domain, potentially decreasing BXA binding but at some cost to the virus. These data implicate E23G/K, alone or combined with I38T, as important markers of reduced BXM susceptibility, and such mutants could emerge and/or transmit among humans. Public Library of Science 2022-07-13 /pmc/articles/PMC9312377/ /pubmed/35830486 http://dx.doi.org/10.1371/journal.ppat.1010698 Text en © 2022 Jones et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Jones, Jeremy C.
Zagribelnyy, Bogdan
Pascua, Philippe Noriel Q.
Bezrukov, Dmitry S.
Barman, Subrata
Okda, Faten
Webby, Richard J.
Ivanenkov, Yan A.
Govorkova, Elena A.
Influenza A virus polymerase acidic protein E23G/K substitutions weaken key baloxavir drug-binding contacts with minimal impact on replication and transmission
title Influenza A virus polymerase acidic protein E23G/K substitutions weaken key baloxavir drug-binding contacts with minimal impact on replication and transmission
title_full Influenza A virus polymerase acidic protein E23G/K substitutions weaken key baloxavir drug-binding contacts with minimal impact on replication and transmission
title_fullStr Influenza A virus polymerase acidic protein E23G/K substitutions weaken key baloxavir drug-binding contacts with minimal impact on replication and transmission
title_full_unstemmed Influenza A virus polymerase acidic protein E23G/K substitutions weaken key baloxavir drug-binding contacts with minimal impact on replication and transmission
title_short Influenza A virus polymerase acidic protein E23G/K substitutions weaken key baloxavir drug-binding contacts with minimal impact on replication and transmission
title_sort influenza a virus polymerase acidic protein e23g/k substitutions weaken key baloxavir drug-binding contacts with minimal impact on replication and transmission
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9312377/
https://www.ncbi.nlm.nih.gov/pubmed/35830486
http://dx.doi.org/10.1371/journal.ppat.1010698
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