title: High content screening identifies decaprenyl-phosphoribose 2' epimerase as a target for intracellular antimycobacterial inhibitors.
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 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.
authors.affiliation: Screening Technologies and Pharmacology, Institut Pasteur Korea, Bundang-gu, Seongnam-si, Gyeonggi-do, Korea.
authors.forename: Thierry
authors.initials: T
authors.lastname: Christophe
authors.forename: Mary
authors.initials: M
authors.lastname: Jackson
authors.forename: Hee Kyoung
authors.initials: HK
authors.lastname: Jeon
authors.forename: Denis
authors.initials: D
authors.lastname: Fenistein
authors.forename: Monica
authors.initials: M
authors.lastname: Contreras-Dominguez
authors.forename: Jaeseung
authors.initials: J
authors.lastname: Kim
authors.forename: Auguste
authors.initials: A
authors.lastname: Genovesio
authors.forename: Jean-Philippe
authors.initials: JP
authors.lastname: Carralot
authors.forename: Fanny
authors.initials: F
authors.lastname: Ewann
authors.forename: Eun Hye
authors.initials: EH
authors.lastname: Kim
authors.forename: Sae Yeon
authors.initials: SY
authors.lastname: Lee
authors.forename: Sunhee
authors.initials: S
authors.lastname: Kang
authors.forename: Min Jung
authors.initials: MJ
authors.lastname: Seo
authors.forename: Eun Jung
authors.initials: EJ
authors.lastname: Park
authors.forename: Henrieta
authors.initials: H
authors.lastname: Skovierová
authors.forename: Ha
authors.initials: H
authors.lastname: Pham
authors.forename: Giovanna
authors.initials: G
authors.lastname: Riccardi
authors.forename: Ji Youn
authors.initials: JY
authors.lastname: Nam
authors.forename: Laurent
authors.initials: L
authors.lastname: Marsollier
authors.forename: Marie
authors.initials: M
authors.lastname: Kempf
authors.forename: Marie-Laure
authors.initials: ML
authors.lastname: Joly-Guillou
authors.forename: Taegwon
authors.initials: T
authors.lastname: Oh
authors.forename: Won Kyung
authors.initials: WK
authors.lastname: Shin
authors.forename: Zaesung
authors.initials: Z
authors.lastname: No
authors.forename: Ulf
authors.initials: U
authors.lastname: Nehrbass
authors.forename: Roland
authors.initials: R
authors.lastname: Brosch
authors.forename: Stewart T
authors.initials: ST
authors.lastname: Cole
authors.forename: Priscille
authors.initials: P
authors.lastname: Brodin
mesh_terms: D000818:Animals; D000904:Antibiotics, Antitubercular / Q000494:pharmacology*; D001549:Benzamides / Q000494:pharmacology; D048708:Cell Growth Processes / Q000187:drug effects; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D055808:Drug Discovery / Q000379:methods*; D006801:Humans; D008264:Macrophages / Q000382:microbiology; D051379:Mice; D008826:Microbial Sensitivity Tests; D018613:Microscopy, Confocal / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000201:enzymology; D025341:Principal Component Analysis; D019998:Racemases and Epimerases / Q000037:antagonists & inhibitors*; D015203:Reproducibility of Results; D013329:Structure-Activity Relationship; D014376:Tuberculosis / Q000188:drug therapy / Q000382:microbiology
chemical_list: D000904:Antibiotics, Antitubercular; D001549:Benzamides; D019998:Racemases and Epimerases