PMID- 35012349 OWN - NLM STAT- MEDLINE DCOM- 20230228 LR - 20230307 IS - 2150-7511 (Electronic) VI - 13 IP - 1 DP - 2022 Feb 22 TI - The Role of Fatty Acid Metabolism in Drug Tolerance of Mycobacterium tuberculosis. PG - e0355921 LID - 10.1128/mbio.03559-21 [doi] LID - e03559-21 AB - Mycobacterium tuberculosis can cocatabolize a range of carbon sources. Fatty acids are among the carbons available inside the host's macrophages. Here, we investigated the metabolic changes of the fatty acid-induced dormancy-like state of M. tuberculosis and its involvement in the acquisition of drug tolerance. We conducted metabolomics profiling using a phosphoenolpyruvate carboxykinase (PEPCK)-deficient M. tuberculosis strain in an acetate-induced dormancy-like state, highlighting an overaccumulation of methylcitrate cycle (MCC) intermediates that correlates with enhanced drug tolerance against isoniazid and bedaquiline. Further metabolomics analyses of two M. tuberculosis mutants, an ICL knockdown (KD) strain and PrpD knockout (KO) strain, each lacking an MCC enzyme-isocitrate lyase (ICL) and 2-methylcitrate dehydratase (PrpD), respectively-were conducted after treatment with antibiotics. The ICL KD strain, which lacks the last enzyme of the MCC, showed an overaccumulation of MCC intermediates and a high level of drug tolerance. The PrpD KO strain, however, failed to accumulate MCC intermediates as it lacks the second step of the MCC and showed only a minor level of drug tolerance compared to the ICL KD mutant and its parental strain (CDC1551). Notably, addition of authentic 2-methylisocitrate, an MCC intermediate, improved the M. tuberculosis drug tolerance against antibiotics even in glycerol medium. Furthermore, wild-type M. tuberculosis displayed levels of drug tolerance when cultured in acetate medium significantly greater than those in glycerol medium. Taken together, the fatty acid-induced dormancy-like state remodels the central carbon metabolism of M. tuberculosis that is functionally relevant to acquisition of M. tuberculosis drug tolerance. IMPORTANCE Understanding the mechanisms underlying M. tuberculosis adaptive strategies to achieve drug tolerance is crucial for the identification of new targets and the development of new drugs. Here, we show that acetate medium triggers a drug-tolerant state in M. tuberculosis when challenged with antituberculosis (anti-TB) drugs. This carbon-induced drug-tolerant state is linked to an accumulation of the methylcitrate cycle (MCC) intermediates, whose role was previously known as a detox pathway for propionate metabolism. Three mutant strains with mutations in gluconeogenesis and MCC were used to investigate the correlation between drug tolerance and the accumulation of MCC metabolites. We herein report a new role of the MCC used to provide a survival advantage to M. tuberculosis as a species against both anti-TB drugs upon specific carbon sources. FAU - Quinonez, Camila G AU - Quinonez CG AUID- ORCID: 0000-0002-1316-2290 AD - Department of Molecular Microbiology and Immunology, Keck School of Medicine, University of Southern California, Los Angeles, California, USA. AD - Department of Life Sciences, Faculty of Science and Technology, University of Westminstergrid.12896.34, London, United Kingdom. FAU - Lee, Jae Jin AU - Lee JJ AD - Department of Molecular Microbiology and Immunology, Keck School of Medicine, University of Southern California, Los Angeles, California, USA. FAU - Lim, Juhyeon AU - Lim J AD - Department of Molecular Microbiology and Immunology, Keck School of Medicine, University of Southern California, Los Angeles, California, USA. FAU - Odell, Mark AU - Odell M AD - Department of Life Sciences, Faculty of Science and Technology, University of Westminstergrid.12896.34, London, United Kingdom. AD - School of Life Sciences, University of Lincoln, Lincoln, United Kingdom. FAU - Lawson, Christopher P AU - Lawson CP AD - Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclydegrid.11984.35, Glasgow, United Kingdom. FAU - Anyogu, Amararachukwu AU - Anyogu A AD - Department of Life Sciences, Faculty of Science and Technology, University of Westminstergrid.12896.34, London, United Kingdom. AD - School of Biomedical Sciences, University of West London, London, United Kingdom. FAU - Raheem, Saki AU - Raheem S AUID- ORCID: 0000-0003-1691-4483 AD - Department of Life Sciences, Faculty of Science and Technology, University of Westminstergrid.12896.34, London, United Kingdom. FAU - Eoh, Hyungjin AU - Eoh H AUID- ORCID: 0000-0001-8774-6400 AD - Department of Molecular Microbiology and Immunology, Keck School of Medicine, University of Southern California, Los Angeles, California, USA. LA - eng GR - R21 AI139386/AI/NIAID NIH HHS/United States GR - R56 AI143870/AI/NIAID NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20220111 PL - United States TA - mBio JT - mBio JID - 101519231 RN - PDC6A3C0OX (Glycerol) RN - 7440-44-0 (Carbon) RN - 0 (Tricarboxylic Acids) RN - 0 (Fatty Acids) RN - 0 (Acetates) SB - IM CIN - Microb Cell. 2022 Feb 28;9(5):123-125. PMID: 35647177 MH - Humans MH - *Mycobacterium tuberculosis/genetics MH - Glycerol/metabolism MH - Carbon/metabolism MH - Tricarboxylic Acids/metabolism MH - *Tuberculosis/microbiology MH - Fatty Acids/metabolism MH - Acetates/metabolism PMC - PMC8749430 OTO - NOTNLM OT - acetate OT - drug tolerance OT - fatty acids OT - metabolomics OT - methylcitrate cycle OT - tuberculosis COIS- The authors declare no conflict of interest. EDAT- 2022/01/12 06:00 MHDA- 2023/03/03 06:00 PMCR- 2022/01/11 CRDT- 2022/01/11 05:43 PHST- 2022/01/11 05:43 [entrez] PHST- 2022/01/12 06:00 [pubmed] PHST- 2023/03/03 06:00 [medline] PHST- 2022/01/11 00:00 [pmc-release] AID - 03559-21 [pii] AID - mbio.03559-21 [pii] AID - 10.1128/mbio.03559-21 [doi] PST - ppublish SO - mBio. 2022 Feb 22;13(1):e0355921. doi: 10.1128/mbio.03559-21. Epub 2022 Jan 11.