title: Enhanced respiration prevents drug tolerance and drug resistance inMycobacterium tuberculosis.
abstract: Persistence, manifested as drug tolerance, represents a significant obstacle to global tuberculosis control. The bactericidal drugs isoniazid and rifampicin kill greater than 99% of exponentially growingMycobacterium tuberculosis(Mtb) cells, but the remaining cells are persisters, cells with decreased metabolic rate, refractory to killing by these drugs, and able to generate drug-resistant mutants. We discovered that the combination of cysteine or other small thiols with either isoniazid or rifampicin prevents the formation of drug-tolerant and drug-resistant cells inMtbcultures. This effect was concentration- and time-dependent, relying on increased oxygen consumption that triggered enhanced production of reactive oxygen species. In infected murine macrophages, the addition ofN-acetylcysteine to isoniazid treatment potentiated the killing ofMtbFurthermore, we demonstrate that the addition of small thiols toMtbdrug treatment shifted the menaquinol/menaquinone balance toward a reduced state that stimulatesMtbrespiration and converts persister cells to metabolically active cells. This prevention of both persister cell formation and drug resistance leads ultimately to mycobacterial cell death. Strategies to enhance respiration and initiate oxidative damage should improve tuberculosis chemotherapies.
authors.affiliation: Howard Hughes Medical Institute, Albert Einstein College of Medicine, Bronx, NY 10461.|Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY 10461.
authors.forename: Catherine
authors.initials: C
authors.lastname: Vilchèze
authors.affiliation: Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY 10461.
authors.forename: Travis
authors.initials: T
authors.lastname: Hartman
authors.affiliation: Howard Hughes Medical Institute, Albert Einstein College of Medicine, Bronx, NY 10461.|Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY 10461.
authors.forename: Brian
authors.identifier: 0000-0003-0880-4487
authors.initials: B
authors.lastname: Weinrick
authors.affiliation: Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY 10461.
authors.forename: Paras
authors.initials: P
authors.lastname: Jain
authors.affiliation: Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY 10461.
authors.forename: Torin R
authors.initials: TR
authors.lastname: Weisbrod
authors.affiliation: Gray Box Biology LLC, New York, NY 10027.
authors.forename: Lawrence W
authors.initials: LW
authors.lastname: Leung
authors.affiliation: Department of Medicine, Center for Emerging and Reemerging Pathogens, New Jersey Medical School, Newark, NJ 07103.
authors.forename: Joel S
authors.initials: JS
authors.lastname: Freundlich
authors.affiliation: Howard Hughes Medical Institute, Albert Einstein College of Medicine, Bronx, NY 10461; jacobsw@hhmi.org.|Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY 10461.
authors.forename: William R
authors.initials: WR
authors.lastname: Jacobs
mesh_terms: D000818:Animals; D000995:Antitubercular Agents / Q000494:pharmacology*; D002460:Cell Line; D053960:DNA Breaks; D024881:Drug Resistance, Bacterial / Q000502:physiology*; D007538:Isoniazid; D008264:Macrophages / Q000378:metabolism / Q000382:microbiology; D051379:Mice; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000502:physiology; D010101:Oxygen Consumption / Q000502:physiology*; D017382:Reactive Oxygen Species; D012293:Rifampin
chemical_list: D000995:Antitubercular Agents; D017382:Reactive Oxygen Species; D007538:Isoniazid; D012293:Rifampin
keywords: drug resistance; mycobacterial persister; oxygen consumption; thiol