Uridine as a potentiator of aminoglycosides through activation of carbohydrate transporters

Aminoglycosides (AGs) are broad-spectrum antibiotics effective against Gram-negative bacteria, with uptake dependent on membrane potential. However, the mechanisms of AG entry remain incompletely understood. Here, we identify a previously undescribed uptake pathway via carbohydrate transporters in E. coli. By deleting or overexpressing 26 carbohydrate transporters, we found that 18 facilitated AG uptake, a mechanism conserved across several Gram-negative ESKAPEE pathogens. Using fluorescent-labeled AGs and flow cytometry, we quantified differential uptake. To enhance AG efficacy, we screened 198 carbon sources for their ability to induce transporter expression using a cmtA-gfp fusion. Uridine emerged as a strong inducer of cmtA and 12 additional AG-importing transporters. Coadministration of uridine considerably improved AG efficacy against clinical and resistant E. coli strains by enhancing drug uptake. This combination also improved outcomes in human blood ex vivo and in a murine urinary tract infection model. Given uridine's clinical safety, it holds promise as an adjuvant to potentiate AG treatment against multidrug-resistant infections.