title: Carbon Sources Tune Antibiotic Susceptibility in Pseudomonas aeruginosa via Tricarboxylic Acid Cycle Control.
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.
authors.affiliation: 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.
authors.forename: Sylvain
authors.initials: S
authors.lastname: Meylan
authors.affiliation: 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.
authors.forename: Caroline B M
authors.initials: CBM
authors.lastname: Porter
authors.affiliation: 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.
authors.forename: Jason H
authors.initials: JH
authors.lastname: Yang
authors.affiliation: Department of Molecular Microbiology and Immunology, Brown University, Providence, RI 02912, USA.
authors.forename: Peter
authors.initials: P
authors.lastname: Belenky
authors.affiliation: 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.
authors.forename: Arnaud
authors.initials: A
authors.lastname: Gutierrez
authors.affiliation: 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.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Lobritz
authors.affiliation: Department of Pediatrics, Massachusetts General Hospital, Boston, MA 02114, USA.
authors.forename: Jihye
authors.initials: J
authors.lastname: Park
authors.affiliation: Department of Pediatrics, Massachusetts General Hospital, Boston, MA 02114, USA.
authors.forename: Sun H
authors.initials: SH
authors.lastname: Kim
authors.affiliation: Department of Pediatrics, Massachusetts General Hospital, Boston, MA 02114, USA; Department of Pediatrics, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Samuel M
authors.initials: SM
authors.lastname: Moskowitz
authors.affiliation: 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.
authors.forename: James J
authors.initials: JJ
authors.lastname: Collins
mesh_terms: D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000494:pharmacology*; D018441:Biofilms / Q000187:drug effects; D002244:Carbon / Q000378:metabolism*; D002952:Citric Acid Cycle / Q000187:drug effects*; D008826:Microbial Sensitivity Tests; D011550:Pseudomonas aeruginosa / Q000187:drug effects* / Q000378:metabolism
chemical_list: D000900:Anti-Bacterial Agents; D002244:Carbon
keywords: LC-MS metabolomics; Pseudomonas aeruginosa; TCA cycle; aminoglycoside susceptibility; biochemical persistence; electron transport chain; fumarate; glyoxylate; respiration