High Content Screening Identifies Decaprenyl-Phosphoribose 2′ Epimerase as a Target for Intracellular Antimycobacterial Inhibitors

PLoS Pathogens

Priscille Brodin¹  Fanny Ewann¹  Monica Contreras-Dominguez¹  Jean-Philippe Carralot¹  Eun Hye Kim¹  Giovanna Riccardi²  Stewart T. Cole³  Marie Kempf⁴  Marie-Laure Joly-Guillou⁴  Laurent Marsollier⁴  Auguste Genovesio⁵  Denis Fenistein⁵  Roland Brosch⁶  Taegwon Oh⁷  Won Kyung Shin⁷  Jaeseung Kim⁸  Sae Yeon Lee⁸  Sunhee Kang⁸  Zaesung No⁸  Min Jung Seo⁸  Eun Jung Park⁸  Ha Pham⁹  Mary Jackson⁹  Henrieta Škovierová⁹  Thierry Christophe^1,10  Ulf Nehrbass^1,10  Ji Youn Nam^1,10  Hee Kyoung Jeon^1,10 

[1] Biology of Intracellular Pathogens Inserm Avenir Group, Institut Pasteur Korea, Bundang-gu, Seongnam-si, Gyeonggi-do, Korea;Dipartimento di Genetica e Microbiologia, Università degli Studi di Pavia, Pavia, Italy;Global Health Institute, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland;Groupe d'Etude des Interactions Hôte Pathogène, Université d'Angers, Angers, France;Image Mining, Institut Pasteur Korea, Bundang-gu, Seongnam-si, Gyeonggi-do, Korea;Institut Pasteur, Integrated Mycobacterial Pathogenomics, Paris, France;International Tuberculosis Research Center, Masan, Korea;Medicinal Chemistry, Institut Pasteur Korea, Bundang-gu, Seongnam-si, Gyeonggi-do, Korea;Mycobacteria Research Laboratories, Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado, United States of America;Screening Technologies and Pharmacology, Institut Pasteur Korea, Bundang-gu, Seongnam-si, Gyeonggi-do, Korea

关键词: Mycobacterium tuberculosis;    Macrophages;    Library screening;    Intracellular pathogens;    Host cells;    Cell walls;    Antibacterials;    Tuberculosis;   

DOI  :  10.1371/journal.ppat.1000645

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

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 2′ 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.

【 授权许可】

CC BY