Carbon Sources Tune Antibiotic Susceptibility in Pseudomonas aeruginosa via Tricarboxylic Acid Cycle Control

Sylvain Meylan; Caroline B M Porter; Jason H Yang; Peter Belenky; Arnaud Gutierrez; Michael A Lobritz; Jihye Park; Sun H Kim; Samuel M Moskowitz; James J Collins

doi:10.1016/j.chembiol.2016.12.015

Carbon Sources Tune Antibiotic Susceptibility in Pseudomonas aeruginosa via Tricarboxylic Acid Cycle Control

Cell Chem Biol. 2017 Feb 16;24(2):195-206. doi: 10.1016/j.chembiol.2016.12.015. Epub 2017 Jan 19.

Authors

Affiliations

¹ Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA; Department of Biological Engineering, Institute for Medical Engineering & Science, Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Division of Infectious Diseases, Brigham and Women's Hospital, Boston, MA 02115, USA; Division of Infectious Diseases, Massachusetts General Hospital, Boston, MA 02114, USA.
² Department of Biological Engineering, Institute for Medical Engineering & Science, Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
³ Department of Molecular Microbiology and Immunology, Brown University, Providence, RI 02912, USA.
⁴ Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA; Department of Biological Engineering, Institute for Medical Engineering & Science, Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Division of Infectious Diseases, Massachusetts General Hospital, Boston, MA 02114, USA.
⁵ Department of Pediatrics, Massachusetts General Hospital, Boston, MA 02114, USA.
⁶ Department of Pediatrics, Massachusetts General Hospital, Boston, MA 02114, USA; Department of Pediatrics, Harvard Medical School, Boston, MA 02115, USA.
⁷ Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA; Department of Biological Engineering, Institute for Medical Engineering & Science, Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Harvard-MIT Program, Health Sciences and Technology, Cambridge, MA 02139, USA. Electronic address: jimjc@mit.edu.

Abstract

Metabolically dormant bacteria present a critical challenge to effective antimicrobial therapy because these bacteria are genetically susceptible to antibiotic treatment but phenotypically tolerant. Such tolerance has been attributed to impaired drug uptake, which can be reversed by metabolic stimulation. Here, we evaluate the effects of central carbon metabolite stimulations on aminoglycoside sensitivity in the pathogen Pseudomonas aeruginosa. We identify fumarate as a tobramycin potentiator that activates cellular respiration and generates a proton motive force by stimulating the tricarboxylic acid (TCA) cycle. In contrast, we find that glyoxylate induces phenotypic tolerance by inhibiting cellular respiration with acetyl-coenzyme A diversion through the glyoxylate shunt, despite drug import. Collectively, this work demonstrates that TCA cycle activity is important for both aminoglycoside uptake and downstream lethality and identifies a potential strategy for potentiating aminoglycoside treatment of P. aeruginosa infections.

Keywords: LC-MS metabolomics; Pseudomonas aeruginosa; TCA cycle; aminoglycoside susceptibility; biochemical persistence; electron transport chain; fumarate; glyoxylate; respiration.

MeSH terms

Anti-Bacterial Agents / chemistry
Anti-Bacterial Agents / pharmacology*
Biofilms / drug effects
Carbon / metabolism*
Citric Acid Cycle / drug effects*
Microbial Sensitivity Tests
Pseudomonas aeruginosa / drug effects*
Pseudomonas aeruginosa / metabolism

Substances

Anti-Bacterial Agents
Carbon

Grants and funding

K99 GM118907/GM/NIGMS NIH HHS/United States