Network analysis reveals differential metabolic functionality in antibiotic-resistantPseudomonas aeruginosa

AbstractMetabolic adaptations accompanying the development of antibiotic resistance in bacteria remain poorly understood. To interrogate this relationship, we profiled the growth of lab-evolved antibiotic-resistant lineages of the opportunistic pathogenPseudomonas aeruginosaacross 190 unique carbon sources. We semi-automatically calculated growth dynamics (maximum growth density, growth rate, and time to mid-exponential phase) of over 2,800 growth curves. These data revealed that the evolution of antibiotic resistance resulted in systems-level changes to growth dynamics and metabolic phenotype. Drug-resistant lineages predominantly displayed decreased growth relative to the ancestral lineage; however, resistant lineages occasionally displayed enhanced growth on certain carbon sources, indicating that adaption to drug can provide a growth advantage in certain environments. A genome-scale metabolic network reconstruction (GENRE) ofP. aeruginosastrain UCBPP-PA14 was paired with whole-genome sequencing data of one of the drug-evolved lineages to predict genes contributing to observed changes in metabolism. Finally, we experimentally validatedin silicopredictions to identify genes mutated in resistantP. aeruginosaaffecting loss of catabolic function. Our results build upon previous mechanistic knowledge of drug-induced metabolic adaptation and provide a framework for the identification of metabolic limitations in antibiotic-resistant pathogens. Robust drug-driven changes in bacterial metabolism have the potential to be exploited to select against antibiotic-resistant populations in chronic infections.