Bioinspired Antimicrobial Strategy: An Extremophile Deep Sea Peptide to Combat Cystic Fibrosis Infections Caused by Pseudomonas aeruginosa and Staphylococcus aureus

Cystic fibrosis (CF)-associated lung infections caused by Pseudomonas aeruginosa (P. aeruginosa) and Staphylococcus aureus (S. aureus) remain difficult to treat due to multidrug resistance and the redox instability of the pulmonary environment, which can impair antibiotic efficacy. In this study, we investigated alvinellacin (ALV), a disulfide-stabilized β-hairpin antimicrobial peptide (AMP) derived from the deep-sea polychaete Alvinella pompejana (A. pompejana), as a potential therapeutic agent naturally adapted to redox-fluctuating conditions. The antibacterial and antibiofilm activities of ALV were evaluated against multidrug-resistant (MDR) clinical isolates under CF-like reducing conditions (6 mM dithiothreitol (DTT)). Circular dichroism (CD) analysis showed that DTT did not alter the β-hairpin secondary structure of ALV, supporting its structural stability in CF-like environments. Mechanistic analyses included pore-forming assay, membrane interaction studies, scanning electron microscopy (SEM), lipid-binding assays, cytotoxicity testing, and resistance induction assays, while in vivo efficacy was assessed using the Galleria mellonella infection model. ALV demonstrated strong bactericidal activity that was maintained in the presence of NaCl or human serum. ALV did not induce bacterial resistance and effectively inhibited early-stage biofilm formation and disrupted preformed biofilms, including those of the clinical isolate, even under reducing conditions. The peptide showed selective permeabilization of bacterial membranes linked to its stronger affinity for bacterial membrane lipids and negligible interaction with host-like membranes, with no observed cytotoxicity. In vivo, ALV significantly improved survival in infected larvae. These findings highlight ALV as a promising redox-resilient antimicrobial candidate for treating MDR CF lung infections.