Molecular Insight into <em>Mycobacterium tuberculosis</em> Resistance to Nitrofuranyl Amides Gained through Metagenomics-like Analysis of Spontaneous Mutants
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 <i>Mycobacterium tuberculosis<...
| Main Authors: | , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-09-01
|
| Series: | Pharmaceuticals |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1424-8247/15/9/1136 |
| _version_ | 1797483766289006592 |
|---|---|
| author | Igor Mokrousov Ivaylo Slavchev Natalia Solovieva Marine Dogonadze Anna Vyazovaya Violeta Valcheva Aleksey Masharsky Olesya Belopolskaya Simeon Dimitrov Viacheslav Zhuravlev Isabel Portugal João Perdigão Georgi M. Dobrikov |
| author_facet | Igor Mokrousov Ivaylo Slavchev Natalia Solovieva Marine Dogonadze Anna Vyazovaya Violeta Valcheva Aleksey Masharsky Olesya Belopolskaya Simeon Dimitrov Viacheslav Zhuravlev Isabel Portugal João Perdigão Georgi M. Dobrikov |
| author_sort | Igor Mokrousov |
| collection | DOAJ |
| 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 <i>Mycobacterium tuberculosis</i> H37Rv. The most active compound <b>11</b> 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 <b>11</b> (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: <i>Rv0224c</i>, <i>fbiC</i>, <i>iniA</i>, and <i>Rv1592c</i>. Gene-set interaction analysis revealed a certain weak interaction for gene pairs <i>Rv1592–Rv1639c</i> and <i>Rv1592–Rv0224c</i>. To conclude, this study experimentally demonstrated a multifaceted primary genetic response of <i>M. tuberculosis</i> to the action of nitrofurans. All three <b>11</b>-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. |
| first_indexed | 2024-03-09T22:51:46Z |
| format | Article |
| id | doaj.art-dfe2d0d6f8744416ae081ad8f58b94f8 |
| institution | Directory Open Access Journal |
| issn | 1424-8247 |
| language | English |
| last_indexed | 2024-03-09T22:51:46Z |
| publishDate | 2022-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceuticals |
| spelling | doaj.art-dfe2d0d6f8744416ae081ad8f58b94f82023-11-23T18:19:18ZengMDPI AGPharmaceuticals1424-82472022-09-01159113610.3390/ph15091136Molecular Insight into <em>Mycobacterium tuberculosis</em> Resistance to Nitrofuranyl Amides Gained through Metagenomics-like Analysis of Spontaneous MutantsIgor Mokrousov0Ivaylo Slavchev1Natalia Solovieva2Marine Dogonadze3Anna Vyazovaya4Violeta Valcheva5Aleksey Masharsky6Olesya Belopolskaya7Simeon Dimitrov8Viacheslav Zhuravlev9Isabel Portugal10João Perdigão11Georgi M. Dobrikov12Laboratory of Molecular Epidemiology and Evolutionary Genetics, St. Petersburg Pasteur Institute, 197101 St. Petersburg, RussiaInstitute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Siences, Acad. G. Bonchev Street, bl. 9, 1113 Sofia, BulgariaSt. Petersburg Research Institute of Phthisiopulmonology, 191036 St. Petersburg, RussiaSt. Petersburg Research Institute of Phthisiopulmonology, 191036 St. Petersburg, RussiaLaboratory of Molecular Epidemiology and Evolutionary Genetics, St. Petersburg Pasteur Institute, 197101 St. Petersburg, RussiaThe Stephan Angeloff Institute of Microbiology, Bulgarian Academy of Sciences, Acad. G. Bonchev Street, bl. 26, 1113 Sofia, BulgariaResource Center “Bio-bank Center”, Research Park of St. Petersburg State University, 198504 St. Petersburg, RussiaResource Center “Bio-bank Center”, Research Park of St. Petersburg State University, 198504 St. Petersburg, RussiaThe Stephan Angeloff Institute of Microbiology, Bulgarian Academy of Sciences, Acad. G. Bonchev Street, bl. 26, 1113 Sofia, BulgariaSt. Petersburg Research Institute of Phthisiopulmonology, 191036 St. Petersburg, RussiaiMed.ULisboa–Instituto de Investigação do Medicamento, Faculdade de Farmácia, Universidade de Lisboa, 1649004 Lisbon, PortugaliMed.ULisboa–Instituto de Investigação do Medicamento, Faculdade de Farmácia, Universidade de Lisboa, 1649004 Lisbon, PortugalInstitute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Siences, Acad. G. Bonchev Street, bl. 9, 1113 Sofia, BulgariaWe 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 <i>Mycobacterium tuberculosis</i> H37Rv. The most active compound <b>11</b> 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 <b>11</b> (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: <i>Rv0224c</i>, <i>fbiC</i>, <i>iniA</i>, and <i>Rv1592c</i>. Gene-set interaction analysis revealed a certain weak interaction for gene pairs <i>Rv1592–Rv1639c</i> and <i>Rv1592–Rv0224c</i>. To conclude, this study experimentally demonstrated a multifaceted primary genetic response of <i>M. tuberculosis</i> to the action of nitrofurans. All three <b>11</b>-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.https://www.mdpi.com/1424-8247/15/9/1136nitrofuranyl amides<i>mycobacterium tuberculosis</i>spontaneous mutagenesiswhole-genome sequencing |
| spellingShingle | Igor Mokrousov Ivaylo Slavchev Natalia Solovieva Marine Dogonadze Anna Vyazovaya Violeta Valcheva Aleksey Masharsky Olesya Belopolskaya Simeon Dimitrov Viacheslav Zhuravlev Isabel Portugal João Perdigão Georgi M. Dobrikov Molecular Insight into <em>Mycobacterium tuberculosis</em> Resistance to Nitrofuranyl Amides Gained through Metagenomics-like Analysis of Spontaneous Mutants Pharmaceuticals nitrofuranyl amides <i>mycobacterium tuberculosis</i> spontaneous mutagenesis whole-genome sequencing |
| title | Molecular Insight into <em>Mycobacterium tuberculosis</em> Resistance to Nitrofuranyl Amides Gained through Metagenomics-like Analysis of Spontaneous Mutants |
| title_full | Molecular Insight into <em>Mycobacterium tuberculosis</em> Resistance to Nitrofuranyl Amides Gained through Metagenomics-like Analysis of Spontaneous Mutants |
| title_fullStr | Molecular Insight into <em>Mycobacterium tuberculosis</em> Resistance to Nitrofuranyl Amides Gained through Metagenomics-like Analysis of Spontaneous Mutants |
| title_full_unstemmed | Molecular Insight into <em>Mycobacterium tuberculosis</em> Resistance to Nitrofuranyl Amides Gained through Metagenomics-like Analysis of Spontaneous Mutants |
| title_short | Molecular Insight into <em>Mycobacterium tuberculosis</em> Resistance to Nitrofuranyl Amides Gained through Metagenomics-like Analysis of Spontaneous Mutants |
| title_sort | molecular insight into em mycobacterium tuberculosis em resistance to nitrofuranyl amides gained through metagenomics like analysis of spontaneous mutants |
| topic | nitrofuranyl amides <i>mycobacterium tuberculosis</i> spontaneous mutagenesis whole-genome sequencing |
| url | https://www.mdpi.com/1424-8247/15/9/1136 |
| work_keys_str_mv | AT igormokrousov molecularinsightintoemmycobacteriumtuberculosisemresistancetonitrofuranylamidesgainedthroughmetagenomicslikeanalysisofspontaneousmutants AT ivayloslavchev molecularinsightintoemmycobacteriumtuberculosisemresistancetonitrofuranylamidesgainedthroughmetagenomicslikeanalysisofspontaneousmutants AT nataliasolovieva molecularinsightintoemmycobacteriumtuberculosisemresistancetonitrofuranylamidesgainedthroughmetagenomicslikeanalysisofspontaneousmutants AT marinedogonadze molecularinsightintoemmycobacteriumtuberculosisemresistancetonitrofuranylamidesgainedthroughmetagenomicslikeanalysisofspontaneousmutants AT annavyazovaya molecularinsightintoemmycobacteriumtuberculosisemresistancetonitrofuranylamidesgainedthroughmetagenomicslikeanalysisofspontaneousmutants AT violetavalcheva molecularinsightintoemmycobacteriumtuberculosisemresistancetonitrofuranylamidesgainedthroughmetagenomicslikeanalysisofspontaneousmutants AT alekseymasharsky molecularinsightintoemmycobacteriumtuberculosisemresistancetonitrofuranylamidesgainedthroughmetagenomicslikeanalysisofspontaneousmutants AT olesyabelopolskaya molecularinsightintoemmycobacteriumtuberculosisemresistancetonitrofuranylamidesgainedthroughmetagenomicslikeanalysisofspontaneousmutants AT simeondimitrov molecularinsightintoemmycobacteriumtuberculosisemresistancetonitrofuranylamidesgainedthroughmetagenomicslikeanalysisofspontaneousmutants AT viacheslavzhuravlev molecularinsightintoemmycobacteriumtuberculosisemresistancetonitrofuranylamidesgainedthroughmetagenomicslikeanalysisofspontaneousmutants AT isabelportugal molecularinsightintoemmycobacteriumtuberculosisemresistancetonitrofuranylamidesgainedthroughmetagenomicslikeanalysisofspontaneousmutants AT joaoperdigao molecularinsightintoemmycobacteriumtuberculosisemresistancetonitrofuranylamidesgainedthroughmetagenomicslikeanalysisofspontaneousmutants AT georgimdobrikov molecularinsightintoemmycobacteriumtuberculosisemresistancetonitrofuranylamidesgainedthroughmetagenomicslikeanalysisofspontaneousmutants |