Characterization of the SARS-CoV-2 Mutation Pattern Generated In Vitro by the Antiviral Action of Lycorine

SARS-CoV-2 persists worldwide, driving the demand for effective antivirals that inhibit replication and limit the emergence of resistant variants. Lycorine, a non-nucleoside inhibitor of SARS-CoV-2 RNA-dependent RNA polymerase, exhibits antiviral activity without direct mutagenic effects. Here, we e...

Descripción completa

Detalles Bibliográficos
Autores principales: Maidana, Silvina Soledad, Romera, Sonia Alejandra, Marandino, Ana, Tau, Rocio Lucia, Shammas, Juan Manuel, Panzera, Yanina, Perez, Ruben
Formato: info:ar-repo/semantics/artículo
Lenguaje:Inglés
Publicado: MDPI 2025
Materias:
Acceso en línea:http://hdl.handle.net/20.500.12123/24774
https://www.mdpi.com/2673-8112/5/11/181
https://doi.org/10.3390/covid5110181
_version_ 1855039004612755456
author Maidana, Silvina Soledad
Romera, Sonia Alejandra
Marandino, Ana
Tau, Rocio Lucia
Shammas, Juan Manuel
Panzera, Yanina
Perez, Ruben
author_browse Maidana, Silvina Soledad
Marandino, Ana
Panzera, Yanina
Perez, Ruben
Romera, Sonia Alejandra
Shammas, Juan Manuel
Tau, Rocio Lucia
author_facet Maidana, Silvina Soledad
Romera, Sonia Alejandra
Marandino, Ana
Tau, Rocio Lucia
Shammas, Juan Manuel
Panzera, Yanina
Perez, Ruben
author_sort Maidana, Silvina Soledad
collection INTA Digital
description SARS-CoV-2 persists worldwide, driving the demand for effective antivirals that inhibit replication and limit the emergence of resistant variants. Lycorine, a non-nucleoside inhibitor of SARS-CoV-2 RNA-dependent RNA polymerase, exhibits antiviral activity without direct mutagenic effects. Here, we examine the occurrence of single-nucleotide variants (SNVs) and insertions/deletions (indels) in SARS-CoV-2 B.1.499 strain during serial passages in Vero cells, comparing lycorine-treated cultures (2.5 and 5 µg/mL) with untreated controls. Whole-genome sequencing was used to assess mutation patterns and frequencies. Lycorine-treated passages displayed greater variant diversity than controls, with fixed mutations mainly affecting non-structural proteins (Nsp3-F1375A, Nsp5-L50F, and Nsp14-G265D) and the envelope protein (E-S6L). A 15-nucleotide deletion in the spike gene (QTQTN motif) occurred in both groups but became fixed only in untreated passages, suggesting negative selection under lycorine pressure. Notably, the L50F mutation in Nsp5, previously linked to nirmatrelvir resistance, was found exclusively in lycorine-treated passages. Additionally, a 1-nucleotide deletion in the accessory gene ORF8, detected only under lycorine treatment, resulted in a frameshift mutation that added four amino acids, potentially altering the protein’s function. Overall, lycorine induces a distinct mutation profile, favoring replication-related variants while suppressing deleterious deletions. These findings suggest potential mechanisms of cross-resistance and highlight the importance of monitoring resistance during clinical use.
format info:ar-repo/semantics/artículo
id INTA24774
institution Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina)
language Inglés
publishDate 2025
publishDateRange 2025
publishDateSort 2025
publisher MDPI
publisherStr MDPI
record_format dspace
spelling INTA247742025-12-26T14:58:00Z Characterization of the SARS-CoV-2 Mutation Pattern Generated In Vitro by the Antiviral Action of Lycorine Maidana, Silvina Soledad Romera, Sonia Alejandra Marandino, Ana Tau, Rocio Lucia Shammas, Juan Manuel Panzera, Yanina Perez, Ruben Coronavirus del Síndrome Respiratorio Agudo Grave 2 Mutación Agentes Antivirales Severe Acute Respiratory Syndrome Coronavirus 2 Mutation Antiviral Agents SARS-CoV-2 Lycorine SARS-CoV-2 persists worldwide, driving the demand for effective antivirals that inhibit replication and limit the emergence of resistant variants. Lycorine, a non-nucleoside inhibitor of SARS-CoV-2 RNA-dependent RNA polymerase, exhibits antiviral activity without direct mutagenic effects. Here, we examine the occurrence of single-nucleotide variants (SNVs) and insertions/deletions (indels) in SARS-CoV-2 B.1.499 strain during serial passages in Vero cells, comparing lycorine-treated cultures (2.5 and 5 µg/mL) with untreated controls. Whole-genome sequencing was used to assess mutation patterns and frequencies. Lycorine-treated passages displayed greater variant diversity than controls, with fixed mutations mainly affecting non-structural proteins (Nsp3-F1375A, Nsp5-L50F, and Nsp14-G265D) and the envelope protein (E-S6L). A 15-nucleotide deletion in the spike gene (QTQTN motif) occurred in both groups but became fixed only in untreated passages, suggesting negative selection under lycorine pressure. Notably, the L50F mutation in Nsp5, previously linked to nirmatrelvir resistance, was found exclusively in lycorine-treated passages. Additionally, a 1-nucleotide deletion in the accessory gene ORF8, detected only under lycorine treatment, resulted in a frameshift mutation that added four amino acids, potentially altering the protein’s function. Overall, lycorine induces a distinct mutation profile, favoring replication-related variants while suppressing deleterious deletions. These findings suggest potential mechanisms of cross-resistance and highlight the importance of monitoring resistance during clinical use. Instituto de Virología Fil: Maidana, Silvina Soledad. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; Argentina Fil: Maidana, Silvina Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Maidana, Silvina Soledad. Universidad del Salvador. Facultad de Ciencias Agrarias y Veterinarias. Instituto de Investigación Veterinaria; Argentina Fil: Romera, Sonia Alejandra. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; Argentina Fil: Romera, Sonia Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Romera, Sonia Alejandra. Universidad del Salvador. Facultad de Ciencias Agrarias y Veterinarias. Instituto de Investigación Veterinaria; Argentina Fil: Marandino, Ana. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; Uruguay Fil: Tau, Rocio Lucia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; Argentina Fil: Tau, Rocio Lucia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Shammas, Juan Manuel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; Argentina Fil: Shammas, Juan Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Panzera, Yanina. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; Uruguay Fil: Pérez, Ruben. Universidad de la República. Facultad de Ciencias. Instituto de Biología. Sección Genética Evolutiva; Uruguay 2025-12-26T14:54:52Z 2025-12-26T14:54:52Z 2025-10 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/24774 https://www.mdpi.com/2673-8112/5/11/181 2673-8112 https://doi.org/10.3390/covid5110181 eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf MDPI COVID 5 (11) : 181. (October 2025)
spellingShingle Coronavirus del Síndrome Respiratorio Agudo Grave 2
Mutación
Agentes Antivirales
Severe Acute Respiratory Syndrome Coronavirus 2
Mutation
Antiviral Agents
SARS-CoV-2
Lycorine
Maidana, Silvina Soledad
Romera, Sonia Alejandra
Marandino, Ana
Tau, Rocio Lucia
Shammas, Juan Manuel
Panzera, Yanina
Perez, Ruben
Characterization of the SARS-CoV-2 Mutation Pattern Generated In Vitro by the Antiviral Action of Lycorine
title Characterization of the SARS-CoV-2 Mutation Pattern Generated In Vitro by the Antiviral Action of Lycorine
title_full Characterization of the SARS-CoV-2 Mutation Pattern Generated In Vitro by the Antiviral Action of Lycorine
title_fullStr Characterization of the SARS-CoV-2 Mutation Pattern Generated In Vitro by the Antiviral Action of Lycorine
title_full_unstemmed Characterization of the SARS-CoV-2 Mutation Pattern Generated In Vitro by the Antiviral Action of Lycorine
title_short Characterization of the SARS-CoV-2 Mutation Pattern Generated In Vitro by the Antiviral Action of Lycorine
title_sort characterization of the sars cov 2 mutation pattern generated in vitro by the antiviral action of lycorine
topic Coronavirus del Síndrome Respiratorio Agudo Grave 2
Mutación
Agentes Antivirales
Severe Acute Respiratory Syndrome Coronavirus 2
Mutation
Antiviral Agents
SARS-CoV-2
Lycorine
url http://hdl.handle.net/20.500.12123/24774
https://www.mdpi.com/2673-8112/5/11/181
https://doi.org/10.3390/covid5110181
work_keys_str_mv AT maidanasilvinasoledad characterizationofthesarscov2mutationpatterngeneratedinvitrobytheantiviralactionoflycorine
AT romerasoniaalejandra characterizationofthesarscov2mutationpatterngeneratedinvitrobytheantiviralactionoflycorine
AT marandinoana characterizationofthesarscov2mutationpatterngeneratedinvitrobytheantiviralactionoflycorine
AT taurociolucia characterizationofthesarscov2mutationpatterngeneratedinvitrobytheantiviralactionoflycorine
AT shammasjuanmanuel characterizationofthesarscov2mutationpatterngeneratedinvitrobytheantiviralactionoflycorine
AT panzerayanina characterizationofthesarscov2mutationpatterngeneratedinvitrobytheantiviralactionoflycorine
AT perezruben characterizationofthesarscov2mutationpatterngeneratedinvitrobytheantiviralactionoflycorine