PerSort Facilitates Characterization and Elimination of Persister Subpopulation in Mycobacteria

ABSTRACT Mycobacterium tuberculosis (MTB) generates phenotypic diversity to persist and survive the harsh conditions encountered during infection. MTB avoids immune effectors and antibacterial killing by entering into distinct physiological states. The surviving cells, persisters, are a major barrie...

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Autores principales:	Vivek Srinivas, Mario L. Arrieta-Ortiz, Amardeep Kaur, Eliza J. R. Peterson, Nitin S. Baliga
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	Mycobacterium phenotypic heterogeneity persisters antibiotic tolerance nutrient starvation Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/f5919ba2d5bc42b1ac69ed43c2af7584
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spelling	oai:doaj.org-article:f5919ba2d5bc42b1ac69ed43c2af75842021-12-02T18:15:47ZPerSort Facilitates Characterization and Elimination of Persister Subpopulation in Mycobacteria10.1128/mSystems.01127-202379-5077https://doaj.org/article/f5919ba2d5bc42b1ac69ed43c2af75842020-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSystems.01127-20https://doaj.org/toc/2379-5077ABSTRACT Mycobacterium tuberculosis (MTB) generates phenotypic diversity to persist and survive the harsh conditions encountered during infection. MTB avoids immune effectors and antibacterial killing by entering into distinct physiological states. The surviving cells, persisters, are a major barrier to the timely and relapse-free treatment of tuberculosis (TB). We present for the first time, PerSort, a method to isolate and characterize persisters in the absence of antibiotic or other pressure. We demonstrate the value of PerSort to isolate translationally dormant cells that preexisted in small numbers within Mycobacterium species cultures growing under optimal conditions but that dramatically increased in proportion under stress conditions. The translationally dormant subpopulation exhibited multidrug tolerance and regrowth properties consistent with those of persister cells. Furthermore, PerSort enabled single-cell transcriptional profiling that provided evidence that the translationally dormant persisters were generated through a variety of mechanisms, including vapC30, mazF, and relA/spoT overexpression. Finally, we demonstrate that notwithstanding the varied mechanisms by which the persister cells were generated, they converge on a similar low-oxygen metabolic state that was reversed through activation of respiration to rapidly eliminate persisters fostered under host-relevant stress conditions. We conclude that PerSort provides a new tool to study MTB persisters, enabling targeted strategies to improve and shorten the treatment of TB. IMPORTANCE Mycobacterium tuberculosis (MTB) persists and survives antibiotic treatments by generating phenotypically heterogeneous drug-tolerant subpopulations. The surviving cells, persisters, are a major barrier to the relapse-free treatment of tuberculosis (TB), which is already killing >1.8 million people every year and becoming deadlier with the emergence of multidrug-resistant strains. This study describes PerSort, a cell sorting method to isolate and characterize, without antibiotic treatment, translationally dormant persisters that preexist in small numbers within Mycobacterium cultures. Characterization of this subpopulation has discovered multiple mechanisms by which mycobacterial persisters emerge and unveiled the physiological basis for their dormant and multidrug-tolerant physiological state. This analysis has discovered that activating oxygen respiratory physiology using l-cysteine eliminates preexisting persister subpopulations, potentiating rapid antibiotic killing of mycobacteria under host-relevant stress. PerSort serves as a new tool to study MTB persisters for enabling targeted strategies to improve and shorten the treatment of TB.Vivek SrinivasMario L. Arrieta-OrtizAmardeep KaurEliza J. R. PetersonNitin S. BaligaAmerican Society for MicrobiologyarticleMycobacteriumphenotypic heterogeneitypersistersantibiotic tolerancenutrient starvationMicrobiologyQR1-502ENmSystems, Vol 5, Iss 6 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Mycobacterium phenotypic heterogeneity persisters antibiotic tolerance nutrient starvation Microbiology QR1-502
spellingShingle	Mycobacterium phenotypic heterogeneity persisters antibiotic tolerance nutrient starvation Microbiology QR1-502 Vivek Srinivas Mario L. Arrieta-Ortiz Amardeep Kaur Eliza J. R. Peterson Nitin S. Baliga PerSort Facilitates Characterization and Elimination of Persister Subpopulation in Mycobacteria
description	ABSTRACT Mycobacterium tuberculosis (MTB) generates phenotypic diversity to persist and survive the harsh conditions encountered during infection. MTB avoids immune effectors and antibacterial killing by entering into distinct physiological states. The surviving cells, persisters, are a major barrier to the timely and relapse-free treatment of tuberculosis (TB). We present for the first time, PerSort, a method to isolate and characterize persisters in the absence of antibiotic or other pressure. We demonstrate the value of PerSort to isolate translationally dormant cells that preexisted in small numbers within Mycobacterium species cultures growing under optimal conditions but that dramatically increased in proportion under stress conditions. The translationally dormant subpopulation exhibited multidrug tolerance and regrowth properties consistent with those of persister cells. Furthermore, PerSort enabled single-cell transcriptional profiling that provided evidence that the translationally dormant persisters were generated through a variety of mechanisms, including vapC30, mazF, and relA/spoT overexpression. Finally, we demonstrate that notwithstanding the varied mechanisms by which the persister cells were generated, they converge on a similar low-oxygen metabolic state that was reversed through activation of respiration to rapidly eliminate persisters fostered under host-relevant stress conditions. We conclude that PerSort provides a new tool to study MTB persisters, enabling targeted strategies to improve and shorten the treatment of TB. IMPORTANCE Mycobacterium tuberculosis (MTB) persists and survives antibiotic treatments by generating phenotypically heterogeneous drug-tolerant subpopulations. The surviving cells, persisters, are a major barrier to the relapse-free treatment of tuberculosis (TB), which is already killing >1.8 million people every year and becoming deadlier with the emergence of multidrug-resistant strains. This study describes PerSort, a cell sorting method to isolate and characterize, without antibiotic treatment, translationally dormant persisters that preexist in small numbers within Mycobacterium cultures. Characterization of this subpopulation has discovered multiple mechanisms by which mycobacterial persisters emerge and unveiled the physiological basis for their dormant and multidrug-tolerant physiological state. This analysis has discovered that activating oxygen respiratory physiology using l-cysteine eliminates preexisting persister subpopulations, potentiating rapid antibiotic killing of mycobacteria under host-relevant stress. PerSort serves as a new tool to study MTB persisters for enabling targeted strategies to improve and shorten the treatment of TB.
format	article
author	Vivek Srinivas Mario L. Arrieta-Ortiz Amardeep Kaur Eliza J. R. Peterson Nitin S. Baliga
author_facet	Vivek Srinivas Mario L. Arrieta-Ortiz Amardeep Kaur Eliza J. R. Peterson Nitin S. Baliga
author_sort	Vivek Srinivas
title	PerSort Facilitates Characterization and Elimination of Persister Subpopulation in Mycobacteria
title_short	PerSort Facilitates Characterization and Elimination of Persister Subpopulation in Mycobacteria
title_full	PerSort Facilitates Characterization and Elimination of Persister Subpopulation in Mycobacteria
title_fullStr	PerSort Facilitates Characterization and Elimination of Persister Subpopulation in Mycobacteria
title_full_unstemmed	PerSort Facilitates Characterization and Elimination of Persister Subpopulation in Mycobacteria
title_sort	persort facilitates characterization and elimination of persister subpopulation in mycobacteria
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/f5919ba2d5bc42b1ac69ed43c2af7584
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PerSort Facilitates Characterization and Elimination of Persister Subpopulation in Mycobacteria

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