(504d) Metabolite-Mediated Elimination of Bacterial Persisters by Aminoglycosides

Bacterial persistence is a phenomenon in which a sub-population of dormant cells (persisters) tolerates antibiotic treatment without being genetically resistant to treatment. Bacterial persisters have been suggested to be an important source of chronic and recurrent infections; however, there is currently no viable method for eliminating persisters. Using a metabolic systems approach, we found that specific metabolic stimuli facilitate killing of both Escherichia coli and Staphylococcus aureus persisters by aminoglycosides. These stimuli specifically caused potentiation by aminoglycoside (mistranslation inducing) antibiotics and did not induce observable potentiation by the fluoroquinolone (DNA damaging) or β?lactam (cell-wall damaging) classes of bactericidal antibiotics, suggesting that stimuli bolster a process specific to aminoglycosides, and do not revert persisters to a normal growth physiology. We demonstrate that potentiation can be explained by increased antibiotic uptake and, additionally, that the increased uptake requires proton-motive force (pmf) generation. Through perturbations to the E. coli metabolic network, we determined that the stimuli promote pmf in persisters by generating the reducing equivalent NADH, primarily from pyruvate dehydrogenase. This work establishes a metabolic-based strategy for eliminating bacterial persisters and demonstrates the importance of metabolic environment to antibiotic treatment.