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Directed evolution of phages in biofilms enhances Pseudomonas aeruginosa control through improved lipopolysaccharide

Luciana Meneses1,2, Lucie Valentová3, Sílvio B Santos1,4

  • 1CEB - Centre of Biological Engineering, University of Minho, Braga, Portugal.

Nature Communications
|November 20, 2025
PubMed

Insights

Bacteriophages adapted to cystic fibrosis biofilms show improved efficacy against Pseudomonas aeruginosa. Mutations enhance phage recognition of diverse bacterial populations, paving the way for targeted phage therapy.

Area of Science:

  • Microbiology
  • Virology
  • Biotechnology

Background:

  • Pseudomonas aeruginosa is a major cause of chronic lung infections in cystic fibrosis (CF) patients.
  • Bacteriophages are potential treatments for antibiotic-resistant infections, but P. aeruginosa biofilms present challenges.
  • Biofilm heterogeneity limits phage effectiveness.

Purpose of the Study:

  • To investigate the adaptive evolution of Pbunavirus phage PE1 to P. aeruginosa biofilms.
  • To enhance phage efficacy in controlling CF-derived P. aeruginosa biofilms.
  • To identify mechanisms for overcoming phage limitations in biofilm environments.

Main Methods:

  • Studied adaptive evolution of phage PE1 to CF-derived P. aeruginosa biofilms.
  • Utilized cryo-electron microscopy (cryo-EM), lipopolysaccharide (LPS) profiling, and adsorption assays.
  • Analyzed mutations in phage tail fiber and baseplate genes.

Main Results:

  • Biofilm-adapted PE1 mutants showed enhanced in vitro efficacy against P. aeruginosa biofilms.
  • Mutants demonstrated improved recognition of diverse bacterial populations within biofilms.
  • Mutations improved phage adsorption and enabled recognition of truncated LPS variants.

Conclusions:

  • Biofilm heterogeneity is a key factor limiting phage therapy effectiveness.
  • Specific phage mutations can overcome barriers to biofilm penetration and control.
  • Genomic targets for engineering phages for CF patients were identified.

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