Uncovering nitroxoline activity spectrum, mode of action and resistance across Gram-negative bacteria

Abstract

Nitroxoline is a bacteriostatic quinoline antibiotic, considered a metal chelator inhibiting the activity of RNA-polymerase1. Its clinical indications are limited to uncomplicated urinary tract infections (UTIs), with a clinical susceptibility breakpoint only available forEscherichia coli2. By testing > 1,000 clinical isolates, here we demonstrate a much broader activity spectrum and species-specific bactericidal activity, including multidrug-resistant Gram-negative bacteria for which therapeutic options are limited due to resistance. By combining systematic genetic and proteomic approaches with direct measurement of intracellular metals, we dissect nitroxoline perturbation of metal homeostasis and unveil additional effects on bacterial physiology. We show that nitroxoline affects outer membrane integrity, synergizing with large-scaffold antibiotics and resensitizing colistin-resistant Enterobacteriaceaein vitroandin vivo. We further characterise resistance mechanisms acrossE. coli,Acinetobacter baumanniiandKlebsiella pneumoniae, recapitulating knownE. coliresistance determinants and uncovering novel and conserved mechanisms across species, demonstrating their common effect on nitroxoline efflux.