Fadi Shehadeh, Biswajit Mishra, Raquel Ferrer-Espada, Anindya Basu, LewisOscar Felix, Charilaos Dellis, Narchonai Ganesan, Li Zhang, Andrew T. Martens, Youlian Goulev, M. C. Sherman, Johan Paulsson, Mandar T. Naik, Paul P. Sotiriadis, Eleftherios Mylonakis
Developing short, stable, and potent antimicrobial peptides is a promising strategy to combat antibiotic resistance and persistence. We present CAMPER (Constraint-driven AMP Engineering with Ranking), a mechanistic artificial intelligence framework that integrates machine learning with biophysical ranking to prioritize membrane-targeting peptides effective against persister and biofilm forms of methicillin-resistant Staphylococcus aureus. We apply CAMPER to identify WP-CAMPER1 (12mer) that kills S. aureus MW2 at a minimal inhibitory concentration of 4 µg/mL. A 2% topical WP-CAMPER1 formulation reduces S. aureus MW2 burden by 2.5 log10 (p < 0.0002) in a murine prophylactic skin infection model, while its D-enantiomer, WP-CAMPER1-d, achieves 1.37 log10 (p < 0.0001) reduction in an established biofilm infection model. Single-cell analysis using a high-throughput microfluidic system shows that WP-CAMPER1-d reduces exponential-phase persisters of S. aureus USA300, and, in a deep-seated murine thigh infection model, decreases stationary-phase S. aureus MW2 persisters by 1.6 log10 (p < 0.0001). This study introduces CAMPER, a mechanistic artificial intelligence platform for designing antimicrobial peptides targeting MRSA. CAMPER identified a stable peptide that eradicates MRSA biofilms and persister cells and was active in mouse infection models.