Transcriptomic interplay betweenAcinetobacter baumannii, human macrophage and polymyxin

Abstract

ABSTRACTOptimization of antibiotic therapy has been hindered by our dearth of understanding on the mechanism of the host-pathogen-drug interactions. Here, we employed dual RNA-sequencing to examine transcriptomic perturbations in response to polymyxin B in a co-culture infection model ofAcinetobacter baumanniiand human macrophages. Our findings revealed that polymyxin B treatment induced significant transcriptomic response in macrophage-interactingA. baumannii, exacerbating bacterial oxidative stress, disrupting metal homeostasis, affecting osmoadaptation, triggering stringent stress response, and influencing pathogenic factors. Moreover, infected macrophages adapt heme catabolism, coagulation cascade, and hypoxia-inducible signaling to confront bacterial invasion. DisruptingrcnB,ompW, andtraR/dksAgenes inA. baumanniiimpairs metal homeostasis, osmotic stress defense and stringent responses, thereby enhancing antibacterial killing by polymyxin. These findings shed light on the global stress adaptations at the network level during host-pathogen-drug interactions, revealing promising therapeutic targets for further investigation.IMPORTANCEIn the context of the development of bacterial resistance during the course of antibiotic therapy, the role of macrophages in shaping bacterial response to antibiotic killing remains enigmatic. Herein we employed dual RNA-sequencing and anin vitrotripartite model to delve into the unexplored transcriptional networks of theAcinetobacter baumannii-macrophage-polymyxin axis. Our findings uncovered the potential synergy between macrophages and polymyxin B which appear to act in co-operation to disrupt multiple stress tolerance mechanisms inA. baumannii. Notably, we discovered the critical roles of bacterial nickel/cobalt homeostasis (rcnBfamily), osmotic stress defense (ompWfamily), and stringent response regulator (traR/dksAC4-type zinc finger) in tolerating the last-line antibiotic polymyxin B. Our findings may lead to potential targets for the development of novel therapeutics against the problematic pathogenA. baumannii.