TY - JOUR
T1 - Rifamycin action on RNA polymerase in antibiotictolerant Mycobacterium tuberculosis results in differentially detectable populations
AU - Saito, Kohta
AU - Warrier, Thulasi
AU - Somersan-Karakaya, Selin
AU - Kaminski, Lina
AU - Mi, Jianjie
AU - Jiang, Xiuju
AU - Park, Suna
AU - Shigyo, Kristi
AU - Gold, Ben
AU - Roberts, Julia
AU - Weber, Elaina
AU - Jacobs, William R.
AU - Nathan, Carl F.
N1 - Funding Information:
We thank Dr. Oksana Ocheretina and the staff of the GHESKIO laboratory in Port-Au-Prince, Haiti, for providing the clinical Mtb strain. We are also thankful to Prof. Michael Glickman at Memorial Sloan Kettering Cancer Center in New York for kindly providing the Mtb Erdman strain and Prof. Christopher Sassetti at the University of Massachusetts Medical School, Worcester, for sharing the Mtb H37Rv-UMass strain. We acknowledge Prof. Sabine Ehrt, Dr. Daniel W. Fitzgerald, and their respective teams for helpful input. We thank Ms. Clara Oromendia for statistical advice via the Clinical and Translational Science Center. This work was supported by the Tri- Institutional TB Research Unit (NIH Grant U19 AI111143-01), NIH Grant T32 AI007613 (to K.S.), NIH Grant K08AI108799 (to S.S.-K.), NIH Grant TBRU U19 AI111276 (to W.R.J. and L.K.), NIH Grant R01 AI026170 (to W.R.J. and L.K.), the Human Frontier Science Program Organization (L.K.), and the Milstein Program in Chemical Biology and Translational Medicine. The Department of Microbiology and Immunology is supported by the William Randolph Hearst Trust.
PY - 2017/6/13
Y1 - 2017/6/13
N2 - Mycobacterium tuberculosis (Mtb) encounters stresses during the pathogenesis and treatment of tuberculosis (TB) that can suppress replication of the bacteria and render them phenotypically tolerant to most available drugs. Where studied, the majority of Mtb in the sputum of most untreated subjects with active TB have been found to be nonreplicating by the criterion that they do not grow as colony-forming units (cfus) when plated on agar. However, these cells are viable because they grow when diluted in liquid media. A method for generating such "differentially detectable" (DD) Mtb in vitro would aid studies of the biology and drug susceptibility of this population, but lack of independent confirmation of reported methods has contributed to skepticism about their existence. Here, we identified confounding artifacts that, when avoided, allowed development of a reliable method of producing cultures of ≥90% DD Mtb in starved cells. We then characterized several drugs according to whether they contribute to the generation of DD Mtb or kill them. Of the agents tested, rifamycins led to DD Mtb generation, an effect lacking in a rifampin-resistant strain with a mutation in rpoB, which encodes the canonical rifampin target, the β subunit of RNA polymerase. In contrast, thioridazine did not generate DD Mtb from starved cells but killed those generated by rifampin.
AB - Mycobacterium tuberculosis (Mtb) encounters stresses during the pathogenesis and treatment of tuberculosis (TB) that can suppress replication of the bacteria and render them phenotypically tolerant to most available drugs. Where studied, the majority of Mtb in the sputum of most untreated subjects with active TB have been found to be nonreplicating by the criterion that they do not grow as colony-forming units (cfus) when plated on agar. However, these cells are viable because they grow when diluted in liquid media. A method for generating such "differentially detectable" (DD) Mtb in vitro would aid studies of the biology and drug susceptibility of this population, but lack of independent confirmation of reported methods has contributed to skepticism about their existence. Here, we identified confounding artifacts that, when avoided, allowed development of a reliable method of producing cultures of ≥90% DD Mtb in starved cells. We then characterized several drugs according to whether they contribute to the generation of DD Mtb or kill them. Of the agents tested, rifamycins led to DD Mtb generation, an effect lacking in a rifampin-resistant strain with a mutation in rpoB, which encodes the canonical rifampin target, the β subunit of RNA polymerase. In contrast, thioridazine did not generate DD Mtb from starved cells but killed those generated by rifampin.
KW - Differentially detectable
KW - Mycobacterium tuberculosis
KW - Phenotypic tolerance
KW - Rifampin
KW - Thioridazine
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U2 - 10.1073/pnas.1705385114
DO - 10.1073/pnas.1705385114
M3 - Article
C2 - 28559332
AN - SCOPUS:85020710199
SN - 0027-8424
VL - 114
SP - E4832-E4840
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 24
ER -