High content screening identifies decaprenyl-phosphoribose 2′ epimerase as a target for intracellular antimycobacterial inhibitors

Thierry Christophe*, Mary Jackson, Kyoung Jeon Hee, Denis Fenistein, Monica Contreras-Dominguez, Jaeseung Kim, Auguste Genovesio, Jean Philippe Carralot, Fanny Ewann, Eun Hye Kim, Sae Yeon Lee, Sunhee Kang, Min Jung Seo, Jung Park Eun, Henrieta Škovierová, Ha Pham, Giovanna Riccardi, Nam Ji Youn, Laurent Marsollier, Marie KempfMarie Laure Joly-Guillou, Taegwon Oh, Kyung Shin Won, Zaesung No, Ulf Nehrbass, Roland Brosch, Stewart T. Cole, Priscille Brodin

*Corresponding author for this work

Research output: Contribution to journalArticleResearchpeer-review

249 Citations (Scopus)

Abstract

A critical feature of Mycobacterium tuberculosis, the causative agent of human tuberculosis (TB), is its ability to survive and multiply within macrophages, making these host cells an ideal niche for persisting microbes. Killing the intracellular tubercle bacilli is a key requirement for efficient tuberculosis treatment, yet identifying potent inhibitors has been hampered by labor-intensive techniques and lack of validated targets. Here, we present the development of a phenotypic cell-based assay that uses automated confocal fluorescence microscopy for high throughput screening of chemicals that interfere with the replication of M. tuberculosis within macrophages. Screening a library of 57,000 small molecules led to the identification of 135 active compounds with potent intracellular anti-mycobacterial efficacy and no host cell toxicity. Among these, the dinitrobenzamide derivatives (DNB) showed high activity against M. tuberculosis, including extensively drug resistant (XDR) strains. More importantly, we demonstrate that incubation of M. tuberculosis with DNB inhibited the formation of both lipoarabinomannan and arabinogalactan, attributable to the inhibition of decaprenyl-phospho-arabinose synthesis catalyzed by the decaprenyl-phosphoribose 29 epimerase DprE1/DprE2. Inhibition of this new target will likely contribute to new therapeutic solutions against emerging XDR-TB. Beyond validating the high throughput/content screening approach, our results open new avenues for finding the next generation of antimicrobials.

Original languageEnglish
Article numbere1000645
JournalPLoS Pathogens
Volume5
Issue number10
DOIs
Publication statusPublished - Oct 2009
Externally publishedYes

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