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Molecular Insight into Mycobacterium tuberculosis Resistance to Nitrofuranyl Amides Gained through Metagenomics-like Analysis of Spontaneous Mutants

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Resumo:We performed synthesis of new nitrofuranyl amides and investigated their anti-TB activity and primary genetic response of mycobacteria through whole-genome sequencing (WGS) of spontaneous resistant mutants. The in vitro activity was assessed on reference strain Mycobacterium tuberculosis H37Rv. The most active compound 11 was used for in vitro selection of spontaneous resistant mutants. The same mutations in six genes were detected in bacterial cultures grown under increased concentrations of 11 (2×, 4×, 8× MIC). The mutant positions were presented as mixed wild type and mutant alleles while increasing the concentration of the compound led to the semi-proportional and significant increase in mutant alleles. The identified genes belong to different categories and pathways. Some of them were previously reported as mediating drug resistance or drug tolerance, and counteracting oxidative and nitrosative stress, in particular: Rv0224c, fbiC, iniA, and Rv1592c. Gene-set interaction analysis revealed a certain weak interaction for gene pairs Rv1592–Rv1639c and Rv1592–Rv0224c. To conclude, this study experimentally demonstrated a multifaceted primary genetic response of M. tuberculosis to the action of nitrofurans. All three 11-treated subcultures independently presented the same six SNPs, which suggests their non-random occurrence and likely causative relationship between compound action and possible resistance mechanism.
Autores principais:Mokrousov, Igor
Outros Autores:Slavchev, Ivaylo; Solovieva, Natalia; Dogonadze, Marine; Vyazovaya, Anna; Valcheva, Violeta; Masharsky, Aleksey; Belopolskaya, Olesya; Dimitrov, Simeon; Zhuravlev, Viacheslav; Portugal, Isabel; Perdigão, João; Dobrikov, Georgi M.
Assunto:nitrofuranyl amides mycobacterium tuberculosis spontaneous mutagenesis whole-genome sequencing
Ano:2022
País:Portugal
Tipo de documento:artigo
Tipo de acesso:acesso aberto
Instituição associada:Universidade de Lisboa
Idioma:inglês
Origem:Repositório da Universidade de Lisboa
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author Mokrousov, Igor
author2 Slavchev, Ivaylo
Solovieva, Natalia
Dogonadze, Marine
Vyazovaya, Anna
Valcheva, Violeta
Masharsky, Aleksey
Belopolskaya, Olesya
Dimitrov, Simeon
Zhuravlev, Viacheslav
Portugal, Isabel
Perdigão, João
Dobrikov, Georgi M.
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author_facet Mokrousov, Igor
Slavchev, Ivaylo
Solovieva, Natalia
Dogonadze, Marine
Vyazovaya, Anna
Valcheva, Violeta
Masharsky, Aleksey
Belopolskaya, Olesya
Dimitrov, Simeon
Zhuravlev, Viacheslav
Portugal, Isabel
Perdigão, João
Dobrikov, Georgi M.
author_role author
contributor_name_str_mv Repositório Científico de Acesso Aberto da ULisboa
country_str PT
creators_json_txt [{\"Person.name\":\"Mokrousov, Igor\",\"Person.identifier.orcid\":\"0000-0001-5924-0576\"},{\"Person.name\":\"Slavchev, Ivaylo\"},{\"Person.name\":\"Solovieva, Natalia\"},{\"Person.name\":\"Dogonadze, Marine\"},{\"Person.name\":\"Vyazovaya, Anna\"},{\"Person.name\":\"Valcheva, Violeta\"},{\"Person.name\":\"Masharsky, Aleksey\"},{\"Person.name\":\"Belopolskaya, Olesya\",\"Person.identifier.orcid\":\"0000-0003-2002-4192\"},{\"Person.name\":\"Dimitrov, Simeon\"},{\"Person.name\":\"Zhuravlev, Viacheslav\",\"Person.identifier.orcid\":\"0000-0001-6906-6225\"},{\"Person.name\":\"Portugal, Isabel\",\"Person.identifier.orcid\":\"0000-0002-7843-0006\"},{\"Person.name\":\"Perdigão, João\",\"Person.identifier.orcid\":\"0000-0002-0339-1305\"},{\"Person.name\":\"Dobrikov, Georgi M.\"}]
datacite.contributors.contributor.contributorName.fl_str_mv Repositório Científico de Acesso Aberto da ULisboa
datacite.creators.creator.creatorName.fl_str_mv Mokrousov, Igor
Slavchev, Ivaylo
Solovieva, Natalia
Dogonadze, Marine
Vyazovaya, Anna
Valcheva, Violeta
Masharsky, Aleksey
Belopolskaya, Olesya
Dimitrov, Simeon
Zhuravlev, Viacheslav
Portugal, Isabel
Perdigão, João
Dobrikov, Georgi M.
datacite.date.Accepted.fl_str_mv 2022-09-12T00:00:00Z
datacite.date.available.fl_str_mv 2023-08-17T17:03:52Z
datacite.date.embargoed.fl_str_mv 2023-08-17T17:03:52Z
datacite.rights.fl_str_mv http://purl.org/coar/access_right/c_abf2
datacite.subjects.subject.fl_str_mv nitrofuranyl amides
mycobacterium tuberculosis
spontaneous mutagenesis
whole-genome sequencing
datacite.titles.title.fl_str_mv Molecular Insight into Mycobacterium tuberculosis Resistance to Nitrofuranyl Amides Gained through Metagenomics-like Analysis of Spontaneous Mutants
dc.contributor.none.fl_str_mv Repositório Científico de Acesso Aberto da ULisboa
dc.creator.none.fl_str_mv Mokrousov, Igor
Slavchev, Ivaylo
Solovieva, Natalia
Dogonadze, Marine
Vyazovaya, Anna
Valcheva, Violeta
Masharsky, Aleksey
Belopolskaya, Olesya
Dimitrov, Simeon
Zhuravlev, Viacheslav
Portugal, Isabel
Perdigão, João
Dobrikov, Georgi M.
dc.date.Accepted.fl_str_mv 2022-09-12T00:00:00Z
dc.date.available.fl_str_mv 2023-08-17T17:03:52Z
dc.date.embargoed.fl_str_mv 2023-08-17T17:03:52Z
dc.format.none.fl_str_mv application/pdf
dc.identifier.none.fl_str_mv http://hdl.handle.net/10451/58917
dc.language.none.fl_str_mv eng
dc.publisher.none.fl_str_mv MDPI
dc.rights.cclincense.fl_str_mv http://creativecommons.org/licenses/by/4.0/
dc.rights.none.fl_str_mv http://purl.org/coar/access_right/c_abf2
dc.subject.none.fl_str_mv nitrofuranyl amides
mycobacterium tuberculosis
spontaneous mutagenesis
whole-genome sequencing
dc.title.fl_str_mv Molecular Insight into Mycobacterium tuberculosis Resistance to Nitrofuranyl Amides Gained through Metagenomics-like Analysis of Spontaneous Mutants
dc.type.none.fl_str_mv http://purl.org/coar/resource_type/c_6501
description We performed synthesis of new nitrofuranyl amides and investigated their anti-TB activity and primary genetic response of mycobacteria through whole-genome sequencing (WGS) of spontaneous resistant mutants. The in vitro activity was assessed on reference strain Mycobacterium tuberculosis H37Rv. The most active compound 11 was used for in vitro selection of spontaneous resistant mutants. The same mutations in six genes were detected in bacterial cultures grown under increased concentrations of 11 (2×, 4×, 8× MIC). The mutant positions were presented as mixed wild type and mutant alleles while increasing the concentration of the compound led to the semi-proportional and significant increase in mutant alleles. The identified genes belong to different categories and pathways. Some of them were previously reported as mediating drug resistance or drug tolerance, and counteracting oxidative and nitrosative stress, in particular: Rv0224c, fbiC, iniA, and Rv1592c. Gene-set interaction analysis revealed a certain weak interaction for gene pairs Rv1592–Rv1639c and Rv1592–Rv0224c. To conclude, this study experimentally demonstrated a multifaceted primary genetic response of M. tuberculosis to the action of nitrofurans. All three 11-treated subcultures independently presented the same six SNPs, which suggests their non-random occurrence and likely causative relationship between compound action and possible resistance mechanism.
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eu_rights_str_mv openAccess
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fulltext.url.fl_str_mv https://repositorio.ulisboa.pt/bitstreams/0b731d5c-6c93-4f93-bbd2-cd1383bd8ad7/download
funding.funder.alternateName_str_mv FCT
FCT
funding.funder.identifier_str_mv http://doi.org/10.13039/501100001871
http://doi.org/10.13039/501100001871
funding.funder.name_str_mv Fundação para a Ciência e a Tecnologia
Fundação para a Ciência e a Tecnologia
funding.name_str_mv 6817 - DCRRNI ID
CEEC IND 2017
id ul_35e5c28414d4aa2700a439ad2a7e4fc2
identifier.url.fl_str_mv http://hdl.handle.net/10451/58917
instacron_str ul
institution Universidade de Lisboa
instname_str Universidade de Lisboa
language eng
network_acronym_str ul
network_name_str Repositório da Universidade de Lisboa
oai_identifier_str oai:repositorio.ulisboa.pt:10451/58917
organization_str_mv urn:organizationAcronym:ul
person_str_mv Mokrousov, Igor
Mokrousov, Igor
http://orcid.org/0000-0001-5924-0576
0000-0001-5924-0576
Slavchev, Ivaylo
Solovieva, Natalia
Dogonadze, Marine
Vyazovaya, Anna
Valcheva, Violeta
Masharsky, Aleksey
Belopolskaya, Olesya
Belopolskaya, Olesya
http://orcid.org/0000-0003-2002-4192
0000-0003-2002-4192
Dimitrov, Simeon
Zhuravlev, Viacheslav
Zhuravlev, Viacheslav
http://orcid.org/0000-0001-6906-6225
0000-0001-6906-6225
Portugal, Isabel
Portugal, Isabel
https://www.ciencia-id.pt/E310-5746-C205
E310-5746-C205
http://orcid.org/0000-0002-7843-0006
0000-0002-7843-0006
Perdigão, João
Perdigão, João
https://www.ciencia-id.pt/991C-F26A-9843
991C-F26A-9843
http://orcid.org/0000-0002-0339-1305
0000-0002-0339-1305
Dobrikov, Georgi M.
publishDate 2022
publisher.none.fl_str_mv MDPI
reponame_str Repositório da Universidade de Lisboa
repository_id_str urn:repositoryAcronym:ul
service_str_mv urn:repositoryAcronym:ul
spelling engMDPIpt_PTWe performed synthesis of new nitrofuranyl amides and investigated their anti-TB activity and primary genetic response of mycobacteria through whole-genome sequencing (WGS) of spontaneous resistant mutants. The in vitro activity was assessed on reference strain Mycobacterium tuberculosis H37Rv. The most active compound 11 was used for in vitro selection of spontaneous resistant mutants. The same mutations in six genes were detected in bacterial cultures grown under increased concentrations of 11 (2×, 4×, 8× MIC). The mutant positions were presented as mixed wild type and mutant alleles while increasing the concentration of the compound led to the semi-proportional and significant increase in mutant alleles. The identified genes belong to different categories and pathways. Some of them were previously reported as mediating drug resistance or drug tolerance, and counteracting oxidative and nitrosative stress, in particular: Rv0224c, fbiC, iniA, and Rv1592c. Gene-set interaction analysis revealed a certain weak interaction for gene pairs Rv1592–Rv1639c and Rv1592–Rv0224c. To conclude, this study experimentally demonstrated a multifaceted primary genetic response of M. tuberculosis to the action of nitrofurans. All three 11-treated subcultures independently presented the same six SNPs, which suggests their non-random occurrence and likely causative relationship between compound action and possible resistance mechanism.application/pdfpt_PTMolecular Insight into Mycobacterium tuberculosis Resistance to Nitrofuranyl Amides Gained through Metagenomics-like Analysis of Spontaneous MutantsPersonalMokrousov, IgorDSpacehttp://dspace.org/items/0ab00fcb-3049-4e7e-b995-8e45b21c43e4DSpacehttp://dspace.org/items/0ab00fcb-3049-4e7e-b995-8e45b21c43e4MokrousovIgorORCIDhttp://orcid.org0000-0001-5924-0576Researcher IDhttps://www.researcherid.comJ-3640-2014Scopus Author IDhttps://www.scopus.com7003916739Slavchev, IvayloSolovieva, NataliaDogonadze, MarineVyazovaya, AnnaValcheva, VioletaMasharsky, AlekseyPersonalBelopolskaya, OlesyaDSpacehttp://dspace.org/items/dac967ad-f99e-4ba9-8962-5adae8765bbfDSpacehttp://dspace.org/items/dac967ad-f99e-4ba9-8962-5adae8765bbfBelopolskayaOlesyaORCIDhttp://orcid.org0000-0003-2002-4192Researcher IDhttps://www.researcherid.comG-9066-2017Scopus Author IDhttps://www.scopus.com55250034500Dimitrov, SimeonPersonalZhuravlev, ViacheslavDSpacehttp://dspace.org/items/0660b61c-4a04-4548-afc3-b84e5e5a1882DSpacehttp://dspace.org/items/0660b61c-4a04-4548-afc3-b84e5e5a1882ZhuravlevViacheslavORCIDhttp://orcid.org0000-0001-6906-6225Researcher IDhttps://www.researcherid.comH-1223-2017Scopus Author IDhttps://www.scopus.com57204401142PersonalPortugal, IsabelDSpacehttp://dspace.org/items/4484da69-843c-4a20-af02-b3c111f84324DSpacehttp://dspace.org/items/4484da69-843c-4a20-af02-b3c111f84324PortugalIsabelCiência IDhttps://www.ciencia-id.ptE310-5746-C205ORCIDhttp://orcid.org0000-0002-7843-0006PersonalPerdigão, JoãoDSpacehttp://dspace.org/items/26de40fc-fc1d-4ff9-a242-3bb398e3ac1eDSpacehttp://dspace.org/items/26de40fc-fc1d-4ff9-a242-3bb398e3ac1eLucas Mota PerdigãoJoão RubenCiência IDhttps://www.ciencia-id.pt991C-F26A-9843ORCIDhttp://orcid.org0000-0002-0339-1305Scopus Author IDhttps://www.scopus.com24723211800Dobrikov, Georgi M.HostingInstitutionOrganizationalRepositório Científico de Acesso Aberto da ULisboae-mailmailto:repositorio@reitoria.ulisboa.ptrepositorio@reitoria.ulisboa.ptISSNIsPartOf1424-8247DOIIsPartOf10.3390/ph150911362023-08-17T17:03:52Z2022-09-122023-01-02T16:43:56Z2022-09-12T00:00:00ZHandlehttp://hdl.handle.net/10451/58917http://purl.org/coar/access_right/c_abf2open accessnitrofuranyl amidesmycobacterium tuberculosisspontaneous mutagenesiswhole-genome sequencing2535951 bytesFundação para a Ciência e a TecnologiaResearch Institute for Medicines6817 - DCRRNI IDCrossref Funder IDhttp://doi.org/10.13039/501100001871Fundação para a Ciência e a TecnologiaNot AvailableCEEC IND 2017Crossref Funder IDhttp://doi.org/10.13039/501100001871literaturehttp://purl.org/coar/resource_type/c_6501journal article2022-09-12http://creativecommons.org/licenses/by/4.0/http://purl.org/coar/access_right/c_abf2application/pdffulltexthttps://repositorio.ulisboa.pt/bitstreams/0b731d5c-6c93-4f93-bbd2-cd1383bd8ad7/downloadPharmaceuticals1591136
spellingShingle Molecular Insight into Mycobacterium tuberculosis Resistance to Nitrofuranyl Amides Gained through Metagenomics-like Analysis of Spontaneous Mutants
Mokrousov, Igor
nitrofuranyl amides
mycobacterium tuberculosis
spontaneous mutagenesis
whole-genome sequencing
status SINGLETON
subject.fl_str_mv nitrofuranyl amides
mycobacterium tuberculosis
spontaneous mutagenesis
whole-genome sequencing
title Molecular Insight into Mycobacterium tuberculosis Resistance to Nitrofuranyl Amides Gained through Metagenomics-like Analysis of Spontaneous Mutants
title_full Molecular Insight into Mycobacterium tuberculosis Resistance to Nitrofuranyl Amides Gained through Metagenomics-like Analysis of Spontaneous Mutants
title_fullStr Molecular Insight into Mycobacterium tuberculosis Resistance to Nitrofuranyl Amides Gained through Metagenomics-like Analysis of Spontaneous Mutants
title_full_unstemmed Molecular Insight into Mycobacterium tuberculosis Resistance to Nitrofuranyl Amides Gained through Metagenomics-like Analysis of Spontaneous Mutants
title_short Molecular Insight into Mycobacterium tuberculosis Resistance to Nitrofuranyl Amides Gained through Metagenomics-like Analysis of Spontaneous Mutants
title_sort Molecular Insight into Mycobacterium tuberculosis Resistance to Nitrofuranyl Amides Gained through Metagenomics-like Analysis of Spontaneous Mutants
topic nitrofuranyl amides
mycobacterium tuberculosis
spontaneous mutagenesis
whole-genome sequencing
topic_facet nitrofuranyl amides
mycobacterium tuberculosis
spontaneous mutagenesis
whole-genome sequencing
url http://hdl.handle.net/10451/58917
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