Chemical Grammar of Accumulation in Drug Resistant Acinetobacter baumannii with Subcellular Resolution

Multidrug-resistant (MDR) PROJECT SUMMARY Acinetobacter baumannii (A. baumannii) is recognized as one of the most urgent global health threats due to its high resistance to nearly all available antibiotics. The rising prevalence of infections caused by this pathogen severely limits treatment options and poses a significant challenge to modern healthcare systems. The primary obstacle in treating A. baumannii infections lies in the bacterium’s unique outer membrane, which acts as a formidable barrier, preventing many antibiotics from reaching their intracellular targets. Despite the critical need for new antibiotics and adjuvants that can overcome this barrier, our understanding of the molecular rules that govern antibiotic accumulation into A. baumannii remains limited. This project aims to address this gap by developing and applying innovative high-throughput screening platforms designed to measure the accumulation of small molecules in A. baumannii with subcellular resolution. Our approach leverages two novel platforms: PAC-MAN (Peptidoglycan Accessibility Click-Mediated AssessmeNt) and CAC-MAN (Cytoplasmic Accessibility Click-Mediated AssessmeNt). These platforms will enable the rapid and detailed analysis of a large library of molecules across multiple A. baumannii strains, including drug-resistant variants. The data generated will be used to identify the chemical properties that facilitate molecular entry and accumulation within these bacterial cells. Additionally, we will conduct a large-scale screen to discover new outer membrane disruptors that can synergize with existing antibiotics, enhancing their efficacy against MDR A. baumannii. By integrating these findings with advanced machine learning algorithms, we aim to establish a comprehensive framework for predicting molecular permeability in A. baumannii. The results of this research will provide critical insights into the development of next-generation antibiotics and adjuvants, addressing a pressing need in the fight against antibiotic resistance and improving outcomes for patients suffering from these difficult-to-treat infections.