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Protective role of Cav-1 in pneumolysin-induced endothelial barrier dysfunction

Pneumolysin (PLY) is a bacterial pore forming toxin and primary virulence factor of Streptococcus pneumonia, a major cause of pneumonia. PLY binds cholesterol-rich domains of the endothelial cell (EC) plasma membrane resulting in pore assembly and increased intracellular (IC) Ca(2+) levels that comp...

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Autores principales: Batori, Robert K., Chen, Feng, Bordan, Zsuzsanna, Haigh, Stephen, Su, Yunchao, Verin, Alexander D., Barman, Scott A., Stepp, David W., Chakraborty, Trinad, Lucas, Rudolf, Fulton, David J. R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9363592/
https://www.ncbi.nlm.nih.gov/pubmed/35967431
http://dx.doi.org/10.3389/fimmu.2022.945656
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author Batori, Robert K.
Chen, Feng
Bordan, Zsuzsanna
Haigh, Stephen
Su, Yunchao
Verin, Alexander D.
Barman, Scott A.
Stepp, David W.
Chakraborty, Trinad
Lucas, Rudolf
Fulton, David J. R.
author_facet Batori, Robert K.
Chen, Feng
Bordan, Zsuzsanna
Haigh, Stephen
Su, Yunchao
Verin, Alexander D.
Barman, Scott A.
Stepp, David W.
Chakraborty, Trinad
Lucas, Rudolf
Fulton, David J. R.
author_sort Batori, Robert K.
collection PubMed
description Pneumolysin (PLY) is a bacterial pore forming toxin and primary virulence factor of Streptococcus pneumonia, a major cause of pneumonia. PLY binds cholesterol-rich domains of the endothelial cell (EC) plasma membrane resulting in pore assembly and increased intracellular (IC) Ca(2+) levels that compromise endothelial barrier integrity. Caveolae are specialized plasmalemma microdomains of ECs enriched in cholesterol. We hypothesized that the abundance of cholesterol-rich domains in EC plasma membranes confers cellular susceptibility to PLY. Contrary to this hypothesis, we found increased PLY-induced IC Ca(2+) following membrane cholesterol depletion. Caveolin-1 (Cav-1) is an essential structural protein of caveolae and its regulation by cholesterol levels suggested a possible role in EC barrier function. Indeed, Cav-1 and its scaffolding domain peptide protected the endothelial barrier from PLY-induced disruption. In loss of function experiments, Cav-1 was knocked-out using CRISPR-Cas9 or silenced in human lung microvascular ECs. Loss of Cav-1 significantly enhanced the ability of PLY to disrupt endothelial barrier integrity. Rescue experiments with re-expression of Cav-1 or its scaffolding domain peptide protected the EC barrier against PLY-induced barrier disruption. Dynamin-2 (DNM2) is known to regulate caveolar membrane endocytosis. Inhibition of endocytosis, with dynamin inhibitors or siDNM2 amplified PLY induced EC barrier dysfunction. These results suggest that Cav-1 protects the endothelial barrier against PLY by promoting endocytosis of damaged membrane, thus reducing calcium entry and PLY-dependent signaling.
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spelling pubmed-93635922022-08-11 Protective role of Cav-1 in pneumolysin-induced endothelial barrier dysfunction Batori, Robert K. Chen, Feng Bordan, Zsuzsanna Haigh, Stephen Su, Yunchao Verin, Alexander D. Barman, Scott A. Stepp, David W. Chakraborty, Trinad Lucas, Rudolf Fulton, David J. R. Front Immunol Immunology Pneumolysin (PLY) is a bacterial pore forming toxin and primary virulence factor of Streptococcus pneumonia, a major cause of pneumonia. PLY binds cholesterol-rich domains of the endothelial cell (EC) plasma membrane resulting in pore assembly and increased intracellular (IC) Ca(2+) levels that compromise endothelial barrier integrity. Caveolae are specialized plasmalemma microdomains of ECs enriched in cholesterol. We hypothesized that the abundance of cholesterol-rich domains in EC plasma membranes confers cellular susceptibility to PLY. Contrary to this hypothesis, we found increased PLY-induced IC Ca(2+) following membrane cholesterol depletion. Caveolin-1 (Cav-1) is an essential structural protein of caveolae and its regulation by cholesterol levels suggested a possible role in EC barrier function. Indeed, Cav-1 and its scaffolding domain peptide protected the endothelial barrier from PLY-induced disruption. In loss of function experiments, Cav-1 was knocked-out using CRISPR-Cas9 or silenced in human lung microvascular ECs. Loss of Cav-1 significantly enhanced the ability of PLY to disrupt endothelial barrier integrity. Rescue experiments with re-expression of Cav-1 or its scaffolding domain peptide protected the EC barrier against PLY-induced barrier disruption. Dynamin-2 (DNM2) is known to regulate caveolar membrane endocytosis. Inhibition of endocytosis, with dynamin inhibitors or siDNM2 amplified PLY induced EC barrier dysfunction. These results suggest that Cav-1 protects the endothelial barrier against PLY by promoting endocytosis of damaged membrane, thus reducing calcium entry and PLY-dependent signaling. Frontiers Media S.A. 2022-07-27 /pmc/articles/PMC9363592/ /pubmed/35967431 http://dx.doi.org/10.3389/fimmu.2022.945656 Text en Copyright © 2022 Batori, Chen, Bordan, Haigh, Su, Verin, Barman, Stepp, Chakraborty, Lucas and Fulton https://creativecommons.org/licenses/by/4.0/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.
spellingShingle Immunology
Batori, Robert K.
Chen, Feng
Bordan, Zsuzsanna
Haigh, Stephen
Su, Yunchao
Verin, Alexander D.
Barman, Scott A.
Stepp, David W.
Chakraborty, Trinad
Lucas, Rudolf
Fulton, David J. R.
Protective role of Cav-1 in pneumolysin-induced endothelial barrier dysfunction
title Protective role of Cav-1 in pneumolysin-induced endothelial barrier dysfunction
title_full Protective role of Cav-1 in pneumolysin-induced endothelial barrier dysfunction
title_fullStr Protective role of Cav-1 in pneumolysin-induced endothelial barrier dysfunction
title_full_unstemmed Protective role of Cav-1 in pneumolysin-induced endothelial barrier dysfunction
title_short Protective role of Cav-1 in pneumolysin-induced endothelial barrier dysfunction
title_sort protective role of cav-1 in pneumolysin-induced endothelial barrier dysfunction
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9363592/
https://www.ncbi.nlm.nih.gov/pubmed/35967431
http://dx.doi.org/10.3389/fimmu.2022.945656
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