Global Assessment of <named-content content-type="genus-species">Mycobacterium avium</named-content> subsp. <italic toggle="yes">hominissuis</italic> Genetic Requirement for Growth and Virulence
ABSTRACT Nontuberculous mycobacterial infections caused by the opportunistic pathogen Mycobacterium avium subsp. hominissuis (MAH) are currently receiving renewed attention due to increased incidence combined with difficult treatment. Insights into the disease-causing mechanisms of this species have...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2019-12-01
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| Series: | mSystems |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/mSystems.00402-19 |
| _version_ | 1818722233147719680 |
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| author | Marte S. Dragset Thomas R. Ioerger Maja Loevenich Markus Haug Niruja Sivakumar Anne Marstad Pere Joan Cardona Geir Klinkenberg Eric J. Rubin Magnus Steigedal Trude H. Flo |
| author_facet | Marte S. Dragset Thomas R. Ioerger Maja Loevenich Markus Haug Niruja Sivakumar Anne Marstad Pere Joan Cardona Geir Klinkenberg Eric J. Rubin Magnus Steigedal Trude H. Flo |
| author_sort | Marte S. Dragset |
| collection | DOAJ |
| description | ABSTRACT Nontuberculous mycobacterial infections caused by the opportunistic pathogen Mycobacterium avium subsp. hominissuis (MAH) are currently receiving renewed attention due to increased incidence combined with difficult treatment. Insights into the disease-causing mechanisms of this species have been hampered by difficulties in genetic manipulation of the bacteria. Here, we identified and sequenced a highly transformable, virulent MAH clinical isolate susceptible to high-density transposon mutagenesis, facilitating global gene disruption and subsequent investigation of MAH gene function. By transposon insertion sequencing (TnSeq) of this strain, we defined the MAH genome-wide genetic requirement for virulence and in vitro growth and organized ∼3,500 identified transposon mutants for hypothesis-driven research. The majority (96%) of the genes we identified as essential for MAH in vitro had a mutual ortholog in the related and highly virulent Mycobacterium tuberculosis (Mtb). However, passaging our library through a mouse model of infection revealed a substantial number (54% of total hits) of novel virulence genes. More than 97% of the MAH virulence genes had a mutual ortholog in Mtb. Finally, we validated novel genes required for successful MAH infection: one encoding a probable major facilitator superfamily (MFS) transporter and another encoding a hypothetical protein located in the immediate vicinity of six other identified virulence genes. In summary, we provide new, fundamental insights into the underlying genetic requirement of MAH for growth and host infection. IMPORTANCE Pulmonary disease caused by nontuberculous mycobacteria is increasing worldwide. The majority of these infections are caused by the Mycobacterium avium complex (MAC), whereof >90% are due to Mycobacterium avium subsp. hominissuis (MAH). Treatment of MAH infections is currently difficult, with a combination of antibiotics given for at least 12 months. To control MAH by improved therapy, prevention, and diagnostics, we need to understand the underlying mechanisms of infection. Here, we provide crucial insights into MAH’s global genetic requirements for growth and infection. We find that the vast majority of genes required for MAH growth and virulence (96% and 97%, respectively) have mutual orthologs in the tuberculosis-causing pathogen M. tuberculosis (Mtb). However, we also find growth and virulence genes specific to MAC species. Finally, we validate novel mycobacterial virulence factors that might serve as future drug targets for MAH-specific treatment or translate to broader treatment of related mycobacterial diseases. |
| first_indexed | 2024-12-17T20:51:22Z |
| format | Article |
| id | doaj.art-7001547c94cf460eb494a3b5fa7ce7b6 |
| institution | Directory Open Access Journal |
| issn | 2379-5077 |
| language | English |
| last_indexed | 2024-12-17T20:51:22Z |
| publishDate | 2019-12-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mSystems |
| spelling | doaj.art-7001547c94cf460eb494a3b5fa7ce7b62022-12-21T21:33:01ZengAmerican Society for MicrobiologymSystems2379-50772019-12-014610.1128/mSystems.00402-19Global Assessment of <named-content content-type="genus-species">Mycobacterium avium</named-content> subsp. <italic toggle="yes">hominissuis</italic> Genetic Requirement for Growth and VirulenceMarte S. Dragset0Thomas R. Ioerger1Maja Loevenich2Markus Haug3Niruja Sivakumar4Anne Marstad5Pere Joan Cardona6Geir Klinkenberg7Eric J. Rubin8Magnus Steigedal9Trude H. Flo10Centre of Molecular Inflammation Research, Norwegian University of Science and Technology, Trondheim, NorwayDepartment of Computer Science and Engineering, Texas A&M University, College Station, Texas, USACentre of Molecular Inflammation Research, Norwegian University of Science and Technology, Trondheim, NorwayCentre of Molecular Inflammation Research, Norwegian University of Science and Technology, Trondheim, NorwayCentre of Molecular Inflammation Research, Norwegian University of Science and Technology, Trondheim, NorwayCentre of Molecular Inflammation Research, Norwegian University of Science and Technology, Trondheim, NorwayTuberculosis Research Unit, Germans Trias i Pujol Research Institute, Badalona, Barcelona, SpainDepartment of Biotechnology and Nanomedicine, SINTEF Materials and Chemistry, Trondheim, NorwayDepartment of Immunology and Infectious Diseases, Harvard T. H. Chan School of Public Health, Boston, Massachusetts, USACentre of Molecular Inflammation Research, Norwegian University of Science and Technology, Trondheim, NorwayCentre of Molecular Inflammation Research, Norwegian University of Science and Technology, Trondheim, NorwayABSTRACT Nontuberculous mycobacterial infections caused by the opportunistic pathogen Mycobacterium avium subsp. hominissuis (MAH) are currently receiving renewed attention due to increased incidence combined with difficult treatment. Insights into the disease-causing mechanisms of this species have been hampered by difficulties in genetic manipulation of the bacteria. Here, we identified and sequenced a highly transformable, virulent MAH clinical isolate susceptible to high-density transposon mutagenesis, facilitating global gene disruption and subsequent investigation of MAH gene function. By transposon insertion sequencing (TnSeq) of this strain, we defined the MAH genome-wide genetic requirement for virulence and in vitro growth and organized ∼3,500 identified transposon mutants for hypothesis-driven research. The majority (96%) of the genes we identified as essential for MAH in vitro had a mutual ortholog in the related and highly virulent Mycobacterium tuberculosis (Mtb). However, passaging our library through a mouse model of infection revealed a substantial number (54% of total hits) of novel virulence genes. More than 97% of the MAH virulence genes had a mutual ortholog in Mtb. Finally, we validated novel genes required for successful MAH infection: one encoding a probable major facilitator superfamily (MFS) transporter and another encoding a hypothetical protein located in the immediate vicinity of six other identified virulence genes. In summary, we provide new, fundamental insights into the underlying genetic requirement of MAH for growth and host infection. IMPORTANCE Pulmonary disease caused by nontuberculous mycobacteria is increasing worldwide. The majority of these infections are caused by the Mycobacterium avium complex (MAC), whereof >90% are due to Mycobacterium avium subsp. hominissuis (MAH). Treatment of MAH infections is currently difficult, with a combination of antibiotics given for at least 12 months. To control MAH by improved therapy, prevention, and diagnostics, we need to understand the underlying mechanisms of infection. Here, we provide crucial insights into MAH’s global genetic requirements for growth and infection. We find that the vast majority of genes required for MAH growth and virulence (96% and 97%, respectively) have mutual orthologs in the tuberculosis-causing pathogen M. tuberculosis (Mtb). However, we also find growth and virulence genes specific to MAC species. Finally, we validate novel mycobacterial virulence factors that might serve as future drug targets for MAH-specific treatment or translate to broader treatment of related mycobacterial diseases.https://journals.asm.org/doi/10.1128/mSystems.00402-19conditionally required genesMycobacterium aviumMycobacterium tuberculosistransposon insertion sequencingvirulence genes |
| spellingShingle | Marte S. Dragset Thomas R. Ioerger Maja Loevenich Markus Haug Niruja Sivakumar Anne Marstad Pere Joan Cardona Geir Klinkenberg Eric J. Rubin Magnus Steigedal Trude H. Flo Global Assessment of <named-content content-type="genus-species">Mycobacterium avium</named-content> subsp. <italic toggle="yes">hominissuis</italic> Genetic Requirement for Growth and Virulence mSystems conditionally required genes Mycobacterium avium Mycobacterium tuberculosis transposon insertion sequencing virulence genes |
| title | Global Assessment of <named-content content-type="genus-species">Mycobacterium avium</named-content> subsp. <italic toggle="yes">hominissuis</italic> Genetic Requirement for Growth and Virulence |
| title_full | Global Assessment of <named-content content-type="genus-species">Mycobacterium avium</named-content> subsp. <italic toggle="yes">hominissuis</italic> Genetic Requirement for Growth and Virulence |
| title_fullStr | Global Assessment of <named-content content-type="genus-species">Mycobacterium avium</named-content> subsp. <italic toggle="yes">hominissuis</italic> Genetic Requirement for Growth and Virulence |
| title_full_unstemmed | Global Assessment of <named-content content-type="genus-species">Mycobacterium avium</named-content> subsp. <italic toggle="yes">hominissuis</italic> Genetic Requirement for Growth and Virulence |
| title_short | Global Assessment of <named-content content-type="genus-species">Mycobacterium avium</named-content> subsp. <italic toggle="yes">hominissuis</italic> Genetic Requirement for Growth and Virulence |
| title_sort | global assessment of named content content type genus species mycobacterium avium named content subsp italic toggle yes hominissuis italic genetic requirement for growth and virulence |
| topic | conditionally required genes Mycobacterium avium Mycobacterium tuberculosis transposon insertion sequencing virulence genes |
| url | https://journals.asm.org/doi/10.1128/mSystems.00402-19 |
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