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This article is part of the Research Topic

Emerging Therapeutic Approaches for Cystic Fibrosis

Original Research ARTICLE Provisionally accepted The full-text will be published soon. Notify me

Front. Pharmacol. | doi: 10.3389/fphar.2018.01330

Use of a primary epithelial cell screening tool to investigate phage therapy in cystic fibrosis

 Stephanie Trend1, 2,  Barbara J. Chang3,  Mark A. O'Dea4, Stephen M. Stick1, 2, 5, 6,  Anthony Kicic1, 2, 5, 6, 7*, - on behalf of WAERP1, - AusREC1, 8, 9 and - AREST CF1, 5, 10, 11
  • 1Telethon Kids Institute, Australia
  • 2School of Biomedical Sciences, Faculty of Health and Medical Sciences, University of Western Australia, Australia
  • 3Marshall Centre for Infectious Diseases Research and Training, Australia
  • 4School of Veterinary and Life Sciences, Murdoch University, Australia
  • 5Department of Respiratory Medicine, Perth Children's Hospital, Australia
  • 6Centre for Cell Therapy and Regenerative Medicine (CCTRM), School of Medicine and Pharmacology, The University of Western Australia, Australia
  • 7Occupation and Environment, School of Public Health, Curtin University, Australia
  • 8Robinson Research Institute (RRI), Australia
  • 9Hunter Medical Research Institute, Australia
  • 10Murdoch Childrens Research Institute, Australia
  • 11Department of Pediatrics, Melbourne Medical School, University of Melbourne, Australia

Antimicrobial-resistant microbes are an increasing threat to human health. In cystic fibrosis (CF), airway infections with Pseudomonas aeruginosa remain a key driver of lung damage. With few new antibiotics on the development horizon, alternative therapeutic approaches are needed against antimicrobial-resistant pathogens. Phage therapy, or the use of viruses that infect bacteria, is one proposed novel therapy to treat bacterial infections. However, the airways are complex microenvironments with unique characteristics that may affect the success of novel therapies. Here, three phages of P. aeruginosa (E79, F116, and one novel clinically-derived isolate, designated P5) were screened for activity against 21 P. aeruginosa strains isolated from children with CF. Of these, phage E79 showed broad antibacterial activity (91% of tested strains sensitive) and was selected for further assessment. E79 genomic DNA was extracted, sequenced and confirmed to contain no bacterial pathogenicity genes. High titre phage preparations were then purified using ion-exchange column chromatography and depleted of bacterial endotoxin. Primary airway epithelial cells derived from children with CF (n=8, age range 0.2–5.5 years, 5 males) or healthy non-CF controls (n=8, age range 2.5–4.0 years, 4 males) were then exposed to purified phage for 48 hours. Levels of inflammatory IL-1, IL-6 and IL-8 cytokine production were measured in culture supernatant by immunoassays and the extent of cellular apoptosis was measured using a ssDNA kit. Cytokine and apoptosis levels were compared between E79-stimulated and unstimulated controls, and, encouragingly, purified preparations of E79 did not stimulate any significant inflammatory cytokine responses or induce apoptosis in primary epithelial cells derived from children with or without CF. Collectively, this study demonstrates the feasibility of utilising pre-clinical in vitro culture models to screen therapeutic candidates, and the potential of E79 as a therapeutic phage candidate in CF.

Keywords: Cystic Fibrosis, phage therapy, preclinical models, airway epithelial cells, Pseudomonas aeruginosa, Infection

Received: 29 Jul 2018; Accepted: 29 Oct 2018.

Edited by:

Florian Lesage, Institut National de la Santé et de la Recherche Médicale (INSERM), France

Reviewed by:

Agnieszka Swiatecka-Urban, University of Pittsburgh, United States
Lobelia Samavati, Wayne State University, United States  

Copyright: © 2018 Trend, Chang, O'Dea, Stick, Kicic, on behalf of WAERP, AusREC and AREST CF. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence: Dr. Anthony Kicic, Telethon Kids Institute, Subiaco, Australia, Anthony.Kicic@telethonkids.org.au