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Bacteriophage therapy is currently resurging as a potential complement/alternative to antibiotic treatment. However, preclinical evaluation lacks streamlined approaches. We here focus on preclinical approaches which have been implemented to assess bacteriophage efficacy against  Pseudomonas  biofilms and infections. Laser interferometry and profilometry were applied to measure biofilm matrix permeability and surface geometry changes, respectively. These biophysical approaches were combined with an advanced Airway Surface Liquid infection model, which mimics  in vitro  the normal and CF lung environments, and an  in vivo Galleria  larvae model. These assays have been implemented to analyze KTN4 (279,593 bp dsDNA genome), a type-IV pili dependent, giant phage resembling phiKZ. Upon contact, KTN4 immediately disrupts the  P. aeruginosa  PAO1 biofilm and reduces pyocyanin and siderophore production. The gentamicin exclusion assay on NuLi-1 and CuFi-1 cell lines revealed the decrease of extracellular bacterial load between 4 and 7 logs and successfully prevents wild-type  Pseudomonas  internalization into CF epithelial cells. These properties and the significant rescue of  Galleria  larvae indicate that giant KTN4 phage is a suitable candidate for  in vivo  phage therapy evaluation for lung infection applications.

A proposed integrated approach for the preclinical evaluation of phage therapy in Pseudomonas infections