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A Multi-stress Model for High Throughput Screening Against Non-replicating Mycobacterium tuberculosis

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Mycobacteria Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1285))

Abstract

Models of non-replication help us understand the biology of persistent Mycobacterium tuberculosis. High throughput screening (HTS) against non-replicating M. tuberculosis may lead to identification of tool compounds that affect pathways on which bacterial survival depends in such states, and to development of drugs that can overcome phenotypic tolerance to conventional antimycobacterial agents, which are mostly active against replicating M. tuberculosis. We describe a multi-stress model of non-replication that mimics some of the microenvironmental conditions that M. tuberculosis faces in the host as adapted for HTS. The model includes acidic pH, mild hypoxia, a flux of nitric oxide and other reactive nitrogen intermediates arising from nitrite at low pH, and low concentrations of a fatty acid (butyrate) as a carbon source.

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Acknowledgements

We are very grateful for the numerous contributions towards the development of the non-replicating assay and critical reading of this chapter: Julia Roberts, Yan Ling, Selin Somersan-Karakaya, Landys Lopez-Quezada, David Little, Maneesh Pingle, Kristin Burns-Huang, and Alfonso Mendoza-Losana, Maria Martinez-Hoyos, and Manuel Marin-Amieva at GSK in Tres Cantos, Spain. This work was supported by the TB Drug Accelerator Program of the Bill and Melinda Gates Foundation and by the Abby and Howard P. Milstein Program in Translational Medicine. The Department of Microbiology and Immunology is supported by the William Randolph Hearst Foundation.

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Authors and Affiliations

  1. Department of Microbiology and Immunology, Weill Cornell Medical College, Belfer 1126, 413 East 69th St., New York, NY, 10065, USA

    Ben Gold

  2. Department of Microbiology and Immunology, Weill Cornell Medical College, Belfer 1150, 413 East 69th St., New York, NY, 10065, USA

    Thulasi Warrier

  3. Department of Microbiology and Immunology, Weill Cornell Medical College, 1300 York Avenue, New York, NY, 10065, USA

    Carl Nathan

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  1. Ben Gold
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  2. Thulasi Warrier
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  3. Carl Nathan
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Correspondence to Ben Gold .

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Editors and Affiliations

  1. Infectious Disease Research Inst (IDRI), Seattle, Washington, USA

    Tanya Parish

  2. Infectious Disease Research Inst (IDRI), Seattle, Washington, USA

    David M. Roberts

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Gold, B., Warrier, T., Nathan, C. (2015). A Multi-stress Model for High Throughput Screening Against Non-replicating Mycobacterium tuberculosis . In: Parish, T., Roberts, D. (eds) Mycobacteria Protocols. Methods in Molecular Biology, vol 1285. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2450-9_18

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  • DOI: https://doi.org/10.1007/978-1-4939-2450-9_18

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2449-3

  • Online ISBN: 978-1-4939-2450-9

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