TY - JOUR
T1 - Genomic characterization of lytic bacteriophages targeting genetically diverse Pseudomonas aeruginosa clinical isolates
AU - Nordstrom, Hayley R.
AU - Evans, Daniel R.
AU - Finney, Amanda G.
AU - Westbrook, Kevin J.
AU - Zamora, Paula F.
AU - Hofstaedter, Casey E.
AU - Yassin, Mohamed H.
AU - Pradhan, Akansha
AU - Iovleva, Alina
AU - Ernst, Robert K.
AU - Bomberger, Jennifer M.
AU - Shields, Ryan K.
AU - Doi, Yohei
AU - Van Tyne, Daria
N1 - Funding Information:
We gratefully acknowledge all members of the Van Tyne lab, and in particular Shu-Ting Cho, for helpful input during the preparation of this manuscript. We also thank Carlos Guerrero-Bustamante and Catherine Armbruster for helpful contributions to the study. Research reported in this publication was supported in part by the National Institute of Allergy and Infectious Diseases of the National Institutes of Health under Award Number UM1AI104681. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. This work was also supported by grants BOMBER19R0, BOMBER21P0, VANTYN21GO, and ZAMORA20F0 from the Cystic Fibrosis Foundation, and by the Department of Medicine at the University of Pittsburgh School of Medicine. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. H.R.N. and D.V.T. conceived the study. M.H.Y. R.K.S. and Y.D. provided wastewater and bacterial isolates for screening. H.R.N. D.R.E. A.G.F. and K.J.W. screened and isolated phages. H.R.N. D.R.E. and D.V.T. performed genomic and phenotypic analyses. C.E.H. A.P. P.F.Z. A.I. R.K.E. and J.M.B. assisted with phenotypic characterization of resistant mutants and biofilm-targeting assays. H.R.N. and D.V.T. wrote the manuscript. All authors reviewed and edited the manuscript. J.B. is a consultant for BiomX, Inc. The other authors declare no competing interests.
Funding Information:
We gratefully acknowledge all members of the Van Tyne lab, and in particular Shu-Ting Cho, for helpful input during the preparation of this manuscript. We also thank Carlos Guerrero-Bustamante and Catherine Armbruster for helpful contributions to the study. Research reported in this publication was supported in part by the National Institute of Allergy and Infectious Diseases of the National Institutes of Health under Award Number UM1AI104681 . The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. This work was also supported by grants BOMBER19R0 , BOMBER21P0 , VANTYN21GO , and ZAMORA20F0 from the Cystic Fibrosis Foundation , and by the Department of Medicine at the University of Pittsburgh School of Medicine . The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Publisher Copyright:
© 2022 The Author(s)
PY - 2022/6/17
Y1 - 2022/6/17
N2 - Pseudomonas aeruginosa infections can be difficult to treat and new therapeutics are needed. Bacteriophage therapy is a promising alternative to traditional antibiotics, but large numbers of isolated and characterized phages are lacking. We collected 23 diverse P. aeruginosa isolates from people with cystic fibrosis (CF) and clinical infections, and used them to screen and isolate over a dozen P. aeruginosa-targeting phages from hospital wastewater. Phages were characterized with genome sequencing, comparative genomics, and lytic activity screening against all 23 bacterial host isolates. We evolved bacterial mutants that were resistant to phage infection for four different phages, and used genome sequencing and functional analysis to study them further. We also tested phages for their ability to kill P. aeruginosa grown in biofilms in vitro and ex vivo on CF airway epithelial cells. Overall, this study demonstrates how systematic genomic and phenotypic characterization can be deployed to develop bacteriophages as precision antibiotics.
AB - Pseudomonas aeruginosa infections can be difficult to treat and new therapeutics are needed. Bacteriophage therapy is a promising alternative to traditional antibiotics, but large numbers of isolated and characterized phages are lacking. We collected 23 diverse P. aeruginosa isolates from people with cystic fibrosis (CF) and clinical infections, and used them to screen and isolate over a dozen P. aeruginosa-targeting phages from hospital wastewater. Phages were characterized with genome sequencing, comparative genomics, and lytic activity screening against all 23 bacterial host isolates. We evolved bacterial mutants that were resistant to phage infection for four different phages, and used genome sequencing and functional analysis to study them further. We also tested phages for their ability to kill P. aeruginosa grown in biofilms in vitro and ex vivo on CF airway epithelial cells. Overall, this study demonstrates how systematic genomic and phenotypic characterization can be deployed to develop bacteriophages as precision antibiotics.
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UR - http://www.scopus.com/inward/citedby.url?scp=85130354027&partnerID=8YFLogxK
U2 - 10.1016/j.isci.2022.104372
DO - 10.1016/j.isci.2022.104372
M3 - Article
AN - SCOPUS:85130354027
SN - 2589-0042
VL - 25
JO - iScience
JF - iScience
IS - 6
M1 - 104372
ER -