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A pipeline to evaluate inhibitors of the Pseudomonas aeruginosa exotoxin U

Pseudomonas aeruginosa has recently been highlighted by the World Health Organisation (WHO) as a major threat with high priority for the development of new therapies. In severe P. aeruginosa infections, the phospholipase activity of the type 3 secretion system toxin, ExoU, induces lysis of target ho...

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Autores principales: Foulkes, Daniel M., McLean, Keri, Zheng, Yalin, Sarsby, Joscelyn, Haneef, Atikah S., Fernig, David G., Winstanley, Craig, Berry, Neil, Kaye, Stephen B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Portland Press Ltd. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7886320/
https://www.ncbi.nlm.nih.gov/pubmed/33459338
http://dx.doi.org/10.1042/BCJ20200780
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author Foulkes, Daniel M.
McLean, Keri
Zheng, Yalin
Sarsby, Joscelyn
Haneef, Atikah S.
Fernig, David G.
Winstanley, Craig
Berry, Neil
Kaye, Stephen B.
author_facet Foulkes, Daniel M.
McLean, Keri
Zheng, Yalin
Sarsby, Joscelyn
Haneef, Atikah S.
Fernig, David G.
Winstanley, Craig
Berry, Neil
Kaye, Stephen B.
author_sort Foulkes, Daniel M.
collection PubMed
description Pseudomonas aeruginosa has recently been highlighted by the World Health Organisation (WHO) as a major threat with high priority for the development of new therapies. In severe P. aeruginosa infections, the phospholipase activity of the type 3 secretion system toxin, ExoU, induces lysis of target host cells and results in the poorest clinical outcomes. We have developed an integrated pipeline to evaluate small molecule inhibitors of ExoU in vitro and in cultured cell models, including a disease-relevant corneal epithelial (HCE-T) scratch and infection model using florescence microscopy and cell viability assays. Compounds Pseudolipasin A, compound A and compound B were effective in vitro inhibitors of ExoU and mitigated P. aeruginosa ExoU-dependent cytotoxicity after infection of HCE-T cells at concentrations as low as 0.5 µM. Addition of the antimicrobial moxifloxacin controlled bacterial load, allowing these assays to be extended from 6 h to 24 h. P. aeruginosa remained cytotoxic to HCE-T cells with moxifloxacin, present at the minimal inhibitory concentration for 24 h, but, when used in combination with either Pseudolipasin A, compound A or compound B, a greater amount of viable cells and scratch healing were observed. Thus, our pipeline provides evidence that ExoU inhibitors could be used in combination with certain antimicrobials as a novel means to treat infections due to ExoU producing P. aeruginosa, as well as the means to identify more potent ExoU inhibitors for future therapeutics.
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spelling pubmed-78863202021-02-23 A pipeline to evaluate inhibitors of the Pseudomonas aeruginosa exotoxin U Foulkes, Daniel M. McLean, Keri Zheng, Yalin Sarsby, Joscelyn Haneef, Atikah S. Fernig, David G. Winstanley, Craig Berry, Neil Kaye, Stephen B. Biochem J Enzymology Pseudomonas aeruginosa has recently been highlighted by the World Health Organisation (WHO) as a major threat with high priority for the development of new therapies. In severe P. aeruginosa infections, the phospholipase activity of the type 3 secretion system toxin, ExoU, induces lysis of target host cells and results in the poorest clinical outcomes. We have developed an integrated pipeline to evaluate small molecule inhibitors of ExoU in vitro and in cultured cell models, including a disease-relevant corneal epithelial (HCE-T) scratch and infection model using florescence microscopy and cell viability assays. Compounds Pseudolipasin A, compound A and compound B were effective in vitro inhibitors of ExoU and mitigated P. aeruginosa ExoU-dependent cytotoxicity after infection of HCE-T cells at concentrations as low as 0.5 µM. Addition of the antimicrobial moxifloxacin controlled bacterial load, allowing these assays to be extended from 6 h to 24 h. P. aeruginosa remained cytotoxic to HCE-T cells with moxifloxacin, present at the minimal inhibitory concentration for 24 h, but, when used in combination with either Pseudolipasin A, compound A or compound B, a greater amount of viable cells and scratch healing were observed. Thus, our pipeline provides evidence that ExoU inhibitors could be used in combination with certain antimicrobials as a novel means to treat infections due to ExoU producing P. aeruginosa, as well as the means to identify more potent ExoU inhibitors for future therapeutics. Portland Press Ltd. 2021-02-12 2021-02-12 /pmc/articles/PMC7886320/ /pubmed/33459338 http://dx.doi.org/10.1042/BCJ20200780 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by/4.0/ This is an open access article published by Portland Press Limited on behalf of the Biochemical Society and distributed under the Creative Commons Attribution License 4.0 (CC BY) (https://creativecommons.org/licenses/by/4.0/) . Open access for this article was enabled by the participation of University of Liverpool in an all-inclusive Read & Publish pilot with Portland Press and the Biochemical Society under a transformative agreement with JISC.
spellingShingle Enzymology
Foulkes, Daniel M.
McLean, Keri
Zheng, Yalin
Sarsby, Joscelyn
Haneef, Atikah S.
Fernig, David G.
Winstanley, Craig
Berry, Neil
Kaye, Stephen B.
A pipeline to evaluate inhibitors of the Pseudomonas aeruginosa exotoxin U
title A pipeline to evaluate inhibitors of the Pseudomonas aeruginosa exotoxin U
title_full A pipeline to evaluate inhibitors of the Pseudomonas aeruginosa exotoxin U
title_fullStr A pipeline to evaluate inhibitors of the Pseudomonas aeruginosa exotoxin U
title_full_unstemmed A pipeline to evaluate inhibitors of the Pseudomonas aeruginosa exotoxin U
title_short A pipeline to evaluate inhibitors of the Pseudomonas aeruginosa exotoxin U
title_sort pipeline to evaluate inhibitors of the pseudomonas aeruginosa exotoxin u
topic Enzymology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7886320/
https://www.ncbi.nlm.nih.gov/pubmed/33459338
http://dx.doi.org/10.1042/BCJ20200780
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