Synthetic Lethality Reveals Mechanisms of <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> Resistance to β-Lactams

ABSTRACT Most β-lactam antibiotics are ineffective against Mycobacterium tuberculosis due to the microbe’s innate resistance. The emergence of multidrug-resistant (MDR) and extensively drug-resistant (XDR) strains has prompted interest to repurpose this class of drugs. To identify the genetic determ...

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Autores principales:	Shichun Lun, David Miranda, Andre Kubler, Haidan Guo, Mariama C. Maiga, Kathryn Winglee, Shaaretha Pelly, William R. Bishai
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2014
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/e2d9cd97723049a59e324f884a5cd039
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spelling	oai:doaj.org-article:e2d9cd97723049a59e324f884a5cd0392021-11-15T15:45:54ZSynthetic Lethality Reveals Mechanisms of <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> Resistance to β-Lactams10.1128/mBio.01767-142150-7511https://doaj.org/article/e2d9cd97723049a59e324f884a5cd0392014-10-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01767-14https://doaj.org/toc/2150-7511ABSTRACT Most β-lactam antibiotics are ineffective against Mycobacterium tuberculosis due to the microbe’s innate resistance. The emergence of multidrug-resistant (MDR) and extensively drug-resistant (XDR) strains has prompted interest to repurpose this class of drugs. To identify the genetic determinants of innate β-lactam resistance, we carried out a synthetic lethality screen on a transposon mutant library for susceptibility to imipenem, a carbapenem β-lactam antibiotic. Mutations in 74 unique genes demonstrated synthetic lethality. The majority of mutations were in genes associated with cell wall biosynthesis. A second quantitative real-time PCR (qPCR)-based synthetic lethality screen of randomly selected mutants confirmed the role of cell wall biosynthesis in β-lactam resistance. The global transcriptional response of the bacterium to β-lactams was investigated, and changes in levels of expression of cell wall biosynthetic genes were identified. Finally, we validated these screens in vivo using the MT1616 transposon mutant, which lacks a functional acyl-transferase gene. Mice infected with the mutant responded to β-lactam treatment with a 100-fold decrease in bacillary lung burden over 4 weeks, while the numbers of organisms in the lungs of mice infected with wild-type bacilli proliferated. These findings reveal a road map of genes required for β-lactam resistance and validate synthetic lethality screening as a promising tool for repurposing existing classes of licensed, safe, well-characterized antimicrobials against tuberculosis. IMPORTANCE The global emergence of multidrug-resistant and extensively drug-resistant M. tuberculosis strains has threatened public health worldwide, yet the pipeline of new tuberculosis drugs under development remains limited. One strategy to cope with the urgent need for new antituberculosis agents is to repurpose existing, approved antibiotics. The carbapenem class of β-lactam antibiotics has been proposed as one such class of drugs. Our study identifies molecular determinants of innate resistance to β-lactam drugs in M. tuberculosis, and we demonstrate that functional loss of one of these genes enables successful treatment of M. tuberculosis with β-lactams in the mouse model.Shichun LunDavid MirandaAndre KublerHaidan GuoMariama C. MaigaKathryn WingleeShaaretha PellyWilliam R. BishaiAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 5, Iss 5 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Shichun Lun David Miranda Andre Kubler Haidan Guo Mariama C. Maiga Kathryn Winglee Shaaretha Pelly William R. Bishai Synthetic Lethality Reveals Mechanisms of <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> Resistance to β-Lactams
description	ABSTRACT Most β-lactam antibiotics are ineffective against Mycobacterium tuberculosis due to the microbe’s innate resistance. The emergence of multidrug-resistant (MDR) and extensively drug-resistant (XDR) strains has prompted interest to repurpose this class of drugs. To identify the genetic determinants of innate β-lactam resistance, we carried out a synthetic lethality screen on a transposon mutant library for susceptibility to imipenem, a carbapenem β-lactam antibiotic. Mutations in 74 unique genes demonstrated synthetic lethality. The majority of mutations were in genes associated with cell wall biosynthesis. A second quantitative real-time PCR (qPCR)-based synthetic lethality screen of randomly selected mutants confirmed the role of cell wall biosynthesis in β-lactam resistance. The global transcriptional response of the bacterium to β-lactams was investigated, and changes in levels of expression of cell wall biosynthetic genes were identified. Finally, we validated these screens in vivo using the MT1616 transposon mutant, which lacks a functional acyl-transferase gene. Mice infected with the mutant responded to β-lactam treatment with a 100-fold decrease in bacillary lung burden over 4 weeks, while the numbers of organisms in the lungs of mice infected with wild-type bacilli proliferated. These findings reveal a road map of genes required for β-lactam resistance and validate synthetic lethality screening as a promising tool for repurposing existing classes of licensed, safe, well-characterized antimicrobials against tuberculosis. IMPORTANCE The global emergence of multidrug-resistant and extensively drug-resistant M. tuberculosis strains has threatened public health worldwide, yet the pipeline of new tuberculosis drugs under development remains limited. One strategy to cope with the urgent need for new antituberculosis agents is to repurpose existing, approved antibiotics. The carbapenem class of β-lactam antibiotics has been proposed as one such class of drugs. Our study identifies molecular determinants of innate resistance to β-lactam drugs in M. tuberculosis, and we demonstrate that functional loss of one of these genes enables successful treatment of M. tuberculosis with β-lactams in the mouse model.
format	article
author	Shichun Lun David Miranda Andre Kubler Haidan Guo Mariama C. Maiga Kathryn Winglee Shaaretha Pelly William R. Bishai
author_facet	Shichun Lun David Miranda Andre Kubler Haidan Guo Mariama C. Maiga Kathryn Winglee Shaaretha Pelly William R. Bishai
author_sort	Shichun Lun
title	Synthetic Lethality Reveals Mechanisms of <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> Resistance to β-Lactams
title_short	Synthetic Lethality Reveals Mechanisms of <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> Resistance to β-Lactams
title_full	Synthetic Lethality Reveals Mechanisms of <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> Resistance to β-Lactams
title_fullStr	Synthetic Lethality Reveals Mechanisms of <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> Resistance to β-Lactams
title_full_unstemmed	Synthetic Lethality Reveals Mechanisms of <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> Resistance to β-Lactams
title_sort	synthetic lethality reveals mechanisms of <named-content content-type="genus-species">mycobacterium tuberculosis</named-content> resistance to β-lactams
publisher	American Society for Microbiology
publishDate	2014
url	https://doaj.org/article/e2d9cd97723049a59e324f884a5cd039
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Synthetic Lethality Reveals Mechanisms of <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> Resistance to β-Lactams

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