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: Puiying A Mak, Srinivasa P S Rao, Mai Ping Tan, Xiuhua Lin, Jason Chyba, Joann Tay, Seow Hwee Ng, Bee Huat Tan, Joseph Cherian, Jeyaraj Duraiswamy, Pablo Bifani, Vivian Lim, Boon Heng Lee, Ngai Ling Ma, David Beer, Pamela Thayalan, Kelli Kuhen, Arnab Chatterjee, Frantisek Supek, Richard Glynne, Jun Zheng, Helena I Boshoff, Clifton E Barry 3rd, Thomas Dick, Kevin Pethe, Luis R Camacho
• Journal: ACS chemical biology (ACS Chem Biol) Vol. 7 Issue 7 Pg. 1190-7 (Jul 20 2012) ISSN: 1554-8937 [Electronic] United States
• PMID: 22500615 (Publication Type: Journal Article, Research Support, N.I.H., Intramural, Research Support, Non-U.S. Gov't)

Chemical References

• Antitubercular Agents
• Adenosine Triphosphate

Topics

• Adenosine Triphosphate (antagonists & inhibitors, physiology)
• Animals
• Antitubercular Agents (chemistry, pharmacology)
• CHO Cells
• Cell Survival (drug effects, physiology)
• Cricetinae
• Cricetulus
• HeLa Cells
• High-Throughput Screening Assays (methods)
• Homeostasis (drug effects, physiology)
• Humans
• Mycobacterium bovis (drug effects, growth & development)
• Mycobacterium tuberculosis (drug effects, growth & development)