Nano-GlycOTAC eradicates Mycobacterium tuberculosis biofilms and rectifies iron metabolism to augment host antimicrobial immunity

Cellulose, as a critical structural component of bacterial biofilms, forms a natural barrier that hinders drug penetration and immune cell invasion, and contributes to persistent tuberculosis (TB) infections and antibiotic resistance. Although Generally Recognized as Safe (GRAS)-affirmed cellulase can hydrolyze cellulose matrix, its clinical application is limited by poor biological availability and anti-biofilm activity. Herein, we developed Nano-GlycOTAC technology, constructing a nano-glycolytic targeting chimera (lactoferrin-cellulase nanoparticles, LCNP) composed of biofilm-binding lactoferrin and cellulase to eradicate biofilm-encased Mycobacterium tuberculosis ( Mtb ). LCNP effectively adhered to biofilms via lactoferrin, released cellulase in response to glutathione (GSH) to degrade cellulose, and subsequently disrupted biofilm and exposed the pathogen. Concurrently, the iron-binding capacity of LCNP blocked Mtb iron uptake and promoted macrophage M1 polarization, thereby enhancing host immunity for bacterial clearance. Consequently, LCNP treatment completely eliminated pulmonary hemorrhage and inflammation in bacteria-infected mice, robustly enhancing the therapeutic efficacy and biosafety of cellulase. In addition, LCNP significantly enhanced biofilm degradation and achieved significant reduction of bacterial load in ex vivo patient samples, including sputum, bronchoalveolar lavage fluid (BALF) and pus. The Nano-GlycOTAC approach combined biofilm disruption, metabolism rectification and immunity activation to normalize Mtb metabolism and remodel host immunity. It emerges as a powerful platform for regulating bacteria fate and activity, and combating biofilm-associated and refractory bacterial infections.