title: A high-throughput screen to identify inhibitors of ATP homeostasis in non-replicating Mycobacterium tuberculosis.
abstract: Growing evidence suggests that the presence of a subpopulation of hypoxic non-replicating, phenotypically drug-tolerant mycobacteria is responsible for the prolonged duration of tuberculosis treatment. The discovery of new antitubercular agents active against this subpopulation may help in developing new strategies to shorten the time of tuberculosis therapy. Recently, the maintenance of a low level of bacterial respiration was shown to be a point of metabolic vulnerability in Mycobacterium tuberculosis. Here, we describe the development of a hypoxic model to identify compounds targeting mycobacterial respiratory functions and ATP homeostasis in whole mycobacteria. The model was adapted to 1,536-well plate format and successfully used to screen over 600,000 compounds. Approximately 800 compounds were confirmed to reduce intracellular ATP levels in a dose-dependent manner in Mycobacterium bovis BCG. One hundred and forty non-cytotoxic compounds with activity against hypoxic non-replicating M. tuberculosis were further validated. The resulting collection of compounds that disrupt ATP homeostasis in M. tuberculosis represents a valuable resource to decipher the biology of persistent mycobacteria.
authors.affiliation: Genomics Institute of the Novartis Research Foundation, San Diego, California 92121, United States.
authors.forename: Puiying A
authors.initials: PA
authors.lastname: Mak
authors.forename: Srinivasa P S
authors.initials: SP
authors.lastname: Rao
authors.forename: Mai
authors.initials: M
authors.lastname: Ping Tan
authors.forename: Xiuhua
authors.initials: X
authors.lastname: Lin
authors.forename: Jason
authors.initials: J
authors.lastname: Chyba
authors.forename: Joann
authors.initials: J
authors.lastname: Tay
authors.forename: Seow Hwee
authors.initials: SH
authors.lastname: Ng
authors.forename: Bee Huat
authors.initials: BH
authors.lastname: Tan
authors.forename: Joseph
authors.initials: J
authors.lastname: Cherian
authors.forename: Jeyaraj
authors.initials: J
authors.lastname: Duraiswamy
authors.forename: Pablo
authors.initials: P
authors.lastname: Bifani
authors.forename: Vivian
authors.initials: V
authors.lastname: Lim
authors.forename: Boon Heng
authors.initials: BH
authors.lastname: Lee
authors.forename: Ngai
authors.initials: N
authors.lastname: Ling Ma
authors.forename: David
authors.initials: D
authors.lastname: Beer
authors.forename: Pamela
authors.initials: P
authors.lastname: Thayalan
authors.forename: Kelli
authors.initials: K
authors.lastname: Kuhen
authors.forename: Arnab
authors.initials: A
authors.lastname: Chatterjee
authors.forename: Frantisek
authors.initials: F
authors.lastname: Supek
authors.forename: Richard
authors.initials: R
authors.lastname: Glynne
authors.forename: Jun
authors.initials: J
authors.lastname: Zheng
authors.forename: Helena I
authors.initials: HI
authors.lastname: Boshoff
authors.forename: Clifton E
authors.initials: CE
authors.lastname: Barry
authors.forename: Thomas
authors.initials: T
authors.lastname: Dick
authors.forename: Kevin
authors.initials: K
authors.lastname: Pethe
authors.forename: Luis R
authors.initials: LR
authors.lastname: Camacho
mesh_terms: D000255:Adenosine Triphosphate / Q000037:antagonists & inhibitors* / Q000502:physiology; D000818:Animals; D000995:Antitubercular Agents / Q000737:chemistry / Q000494:pharmacology*; D016466:CHO Cells; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D006224:Cricetinae; D003412:Cricetulus; D006367:HeLa Cells; D057166:High-Throughput Screening Assays / Q000379:methods*; D006706:Homeostasis / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D009163:Mycobacterium bovis / Q000187:drug effects / Q000254:growth & development; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000254:growth & development*
chemical_list: D000995:Antitubercular Agents; D000255:Adenosine Triphosphate