Molecular response to the non-lytic peptide bac7 (1-35) triggers disruption of Klebsiella pneumoniae biofilm
Klebsiella pneumoniae is becoming increasingly difficult to treat as multidrug-resistant (MDR) strains become more prevalent. The formation of biofilm heightens this threat by embedding bacterial cells in a polysaccharide-rich matrix that limits antibiotic penetration. Here we dissect the anti-biofilm bovine host-defense cathelicidin peptide fragment bac7 (1-35), exploring its anti-biofilm mechanism, evaluating its ability to curb dissemination of hypervirulent K. pneumoniae, and testing its breadth of activity against diverse clinical isolates. Transcriptomic profiling revealed that bac7(1-35) simultaneously compromises the bacterial membrane and inhibits ribosomal function, a dual assault that precipitates rapid biofilm collapse and blocks bacterial spread. Further, the peptide eradicated biofilms produced by the strongest MDR clinical isolates in the Multidrug-Resistant Organism Repository and Surveillance Network (MRSN) diversity panel. Although bac7 (1-35) kills bacterial cells via a cytosolic mechanism, membrane interaction profiles varied among MRSN isolates, correlating with differential peptide translocation. In a delayed-treatment murine skin-abscess model, bac7 (1-35) halted in vivo dissemination of the hypervirulent strain NTUH-K2044. Collectively, these results delineate a multifaceted mode of action for bac7 (1-35) and underscore its therapeutic promise against biofilm-associated MDR K. pneumoniae infections. Author SummaryKlebsiella pneumoniae is a top-priority pathogen for new therapies, with many strains already approaching pan-drug resistant status. Biofilm formation further complicates treatment, yet biofilm-active therapeutics have not reached the clinic, in part because we still lack a detailed understanding of how to disrupt these impenetrable structures. Antimicrobial peptides are promising candidates and have shown biofilm-disruption potential. Here we provide mechanistic insight into how a host defense peptide dismantles pre-formed K. pneumoniae biofilms. We find that the peptides dual targeting of bacterial membranes and ribosomes triggers dispersal from the biofilm state and concomitantly downregulates factors required for surface attachment and extracellular matrix production. This mechanism involves a protein that, to our knowledge, has not been characterized in K. pneumoniae. Our findings reveal a switch that can be leveraged to reprogram biofilm maintenance toward dispersal in K. pneumoniae, advancing the path to peptide-based antibiofilm therapeutics.