A dual-targeting lipid nanoparticle delivery system for precision phage therapy against multidrug-resistant Gram-negative bacteria

The escalating crisis of antimicrobial resistance, particularly among Gram-negative bacteria, demands innovative therapeutic strategies. Here, we present a dual-targeting lipid nanoparticle-chimeric phage (AbLCP) system engineered to combat multidrug-resistant (MDR) Gram-negative bacteria. Our design integrates a genetically modified M13 phage displaying anti-lipopolysaccharide (LPS) antibodies with polymyxin B conjugation, which is further encapsulated within anti-LPS antibody-functionalized lipid nanoparticles. This system overcomes the fundamental limitations of conventional phage therapy, enabling efficient cellular internalization, endosomal escape, and potent intracellular bactericidal activity. In murine models of Pseudomonas aeruginosa infection, AbLCP achieved superior bacterial clearance and up to 10³-fold reduction in bacterial burden at a 10³-fold lower effective dose than free polymyxin B, with 67-fold selective accumulation at infection sites, and attenuated systemic inflammation and preservation of gut microbiota. Mechanistically, AbLCP neutralized LPS to suppress inflammation and induced a lethal bacterial stress response. This work establishes a transformative nanophage platform that merges synthetic biology with nanomedicine, offering a promising strategy against MDR bacterial infections.