A proposed integrated approach for the preclinical evaluation of phage therapy in Pseudomonas infections.
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.
Department of Pathogen Biology and Immunology, Institute of Genetics and Microbiology, University of Wroclaw, Wroclaw, Poland.|Laboratory of Gene Technology, KULeuven, Leuven, Belgium.
Katarzyna
K
Danis-Wlodarczyk
Laboratory of Gene Technology, KULeuven, Leuven, Belgium.
Dieter
D
Vandenheuvel
Laboratory of Gene Technology, KULeuven, Leuven, Belgium.
Ho Bin
HB
Jang
Laboratory of Gene Technology, KULeuven, Leuven, Belgium.|Laboratory of Applied Biotechnology, Department of Applied Biosciences, Ghent University, Ghent, Belgium.
Yves
Y
Briers
Department of Pathogen Biology and Immunology, Institute of Genetics and Microbiology, University of Wroclaw, Wroclaw, Poland.
Tomasz
T
Olszak
Department of Microbiology, Institute of Biology, The Jan Kochanowski University in Kielce, Kielce, Poland.
Michal
M
Arabski
Department of Molecular Physics, Institute of Physics, The Jan Kochanowski University in Kielce, Kielce, Poland.
Slawomir
S
Wasik
Department of Astrophysics, Institute of Physics, The Jan Kochanowski University in Kielce, Kielce, Poland.
Marcin
M
Drabik
National Children Research Centre, Dublin, Ireland.|Department of Molecular Medicine, Royal College of Surgeons in Ireland, Education and Research Centre, Beaumont Hospital, Dublin, Ireland.
Gerard
G
Higgins
Department of Molecular Medicine, Royal College of Surgeons in Ireland, Education and Research Centre, Beaumont Hospital, Dublin, Ireland.
Jean
J
Tyrrell
Department of Molecular Medicine, Royal College of Surgeons in Ireland, Education and Research Centre, Beaumont Hospital, Dublin, Ireland.
Brian J
BJ
Harvey
Biomedical Research Institute and Transnationale Universiteit Limburg, School of Life Sciences, Hasselt University, Diepenbeek, Belgium.
Jean-Paul
JP
Noben
Laboratory of Gene Technology, KULeuven, Leuven, Belgium.
Rob
R
Lavigne
Department of Pathogen Biology and Immunology, Institute of Genetics and Microbiology, University of Wroclaw, Wroclaw, Poland.
Zuzanna
Z
Drulis-Kawa
D000818:Animals; D058491:Bacterial Load; D018441:Biofilms; D002460:Cell Line; D003550:Cystic Fibrosis / Q000473:pathology; D004195:Disease Models, Animal; D004847:Epithelial Cells / Q000821:virology; D005839:Gentamicins / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009036:Moths / Q000382:microbiology; D009154:Mutation; D000071059:Phage Therapy / Q000379:methods*; D011552:Pseudomonas Infections / Q000628:therapy*; D017105:Pseudomonas Phages / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000472:pathogenicity / Q000821:virology; D014764:Viral Proteins / Q000737:chemistry
D005839:Gentamicins; D014764:Viral Proteins