Minimal Gene Signatures Enable High-Accuracy Prediction of Antibiotic Resistance in

Antimicrobial resistance (AMR) in Pseudomonas aeruginosa poses a critical global health challenge, with current diagnostics relying on slow, culture-based methods. Here, we present a ML framework leveraging transcriptomic data to predict antibiotic resistance with high accuracy. We applied a genetic algorithm to 414 clinical isolates to identify minimal, highly predictive gene sets (~35-40 genes) distinguishing resistant from susceptible strains for meropenem, ciprofloxacin, tobramycin, and ceftazidime. Automated ML classifiers trained on these sets achieved accuracies of 96-99% on test data (F1 scores: 0.93-0.99), surpassing clinical deployment thresholds. Multiple distinct, non-overlapping gene subsets exhibited comparable performance, indicating that resistance acquisition broadly impacts the expression of diverse regulatory and metabolic genes. Comparison with known resistance markers from CARD and operon annotations revealed a substantial number of previously unannotated clusters, highlighting significant knowledge gaps in current AMR understanding. Mapping these genes onto independently modulated gene sets (iModulons) revealed transcriptional adaptations across diverse genetic regions. Overall, this study presents a streamlined machine-learning workflow for transcriptomic data and offers a pathway toward rapid diagnostics and personalized treatment strategies against AMR.