The pH-responsive PacC transcription factor of Aspergillus fumigatus governs epithelial entry and tissue invasion during pulmonary aspergillosis

Destruction of the pulmonary epithelium is a major feature of lung diseases caused by the mould pathogen Aspergillus fumigatus. Although it is widely postulated that tissue invasion is governed by fungal proteases, A. fumigatus mutants lacking individual or multiple enzymes remain fully invasive, su...

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Autores: Bertuzzi, Margherita, Schrettl, Markus, Alcazar-Fuoli, Laura, Cairns, Timothy C, Muñoz, Alberto, Walker, Louise A, Herbst, Susanne, Safari, Maryam, Cheverton, Angela M, Chen, Dan, Liu, Hong, Saijo, Shinobu, Fedorova, Natalie D, Armstrong-James, Darius, Munro, Carol A, Read, Nick D, Filler, Scott G, Espeso, Eduardo A, Nierman, William C, Haas, Hubertus, Bignell, Elaine M
Tipo de recurso: artículo
Fecha de publicación:2014
País:España
Institución:Instituto de Salud Carlos III (ISCIII)
Repositorio:Repisalud
Idioma:inglés
OAI Identifier:oai:repisalud.isciii.es:20.500.12105/6837
Acceso en línea:http://hdl.handle.net/20.500.12105/6837
Access Level:acceso abierto
Palabra clave:Animals
Aspergillus fumigatus
Epithelial Cells
Fungal Proteins
Hydrogen-Ion Concentration
Mice
Pulmonary Aspergillosis
Transcription Factors
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spelling The pH-responsive PacC transcription factor of Aspergillus fumigatus governs epithelial entry and tissue invasion during pulmonary aspergillosisBertuzzi, MargheritaSchrettl, MarkusAlcazar-Fuoli, LauraCairns, Timothy CMuñoz, AlbertoWalker, Louise AHerbst, SusanneSafari, MaryamCheverton, Angela MChen, DanLiu, HongSaijo, ShinobuFedorova, Natalie DArmstrong-James, DariusMunro, Carol ARead, Nick DFiller, Scott GEspeso, Eduardo ANierman, William CHaas, HubertusBignell, Elaine MAnimalsAspergillus fumigatusEpithelial CellsFungal ProteinsHydrogen-Ion ConcentrationMicePulmonary AspergillosisTranscription FactorsDestruction of the pulmonary epithelium is a major feature of lung diseases caused by the mould pathogen Aspergillus fumigatus. Although it is widely postulated that tissue invasion is governed by fungal proteases, A. fumigatus mutants lacking individual or multiple enzymes remain fully invasive, suggesting a concomitant requirement for other pathogenic activities during host invasion. In this study we discovered, and exploited, a novel, tissue non-invasive, phenotype in A. fumigatus mutants lacking the pH-responsive transcription factor PacC. Our study revealed a novel mode of epithelial entry, occurring in a cell wall-dependent manner prior to protease production, and via the Dectin-1 β-glucan receptor. ΔpacC mutants are defective in both contact-mediated epithelial entry and protease expression, and significantly attenuated for pathogenicity in leukopenic mice. We combined murine infection modelling, in vivo transcriptomics, and in vitro infections of human alveolar epithelia, to delineate two major, and sequentially acting, PacC-dependent processes impacting epithelial integrity in vitro and tissue invasion in the whole animal. We demonstrate that A. fumigatus spores and germlings are internalised by epithelial cells in a contact-, actin-, cell wall- and Dectin-1 dependent manner and ΔpacC mutants, which aberrantly remodel the cell wall during germinative growth, are unable to gain entry into epithelial cells, both in vitro and in vivo. We further show that PacC acts as a global transcriptional regulator of secreted molecules during growth in the leukopenic mammalian lung, and profile the full cohort of secreted gene products expressed during invasive infection. Our study reveals a combinatorial mode of tissue entry dependent upon sequential, and mechanistically distinct, perturbations of the pulmonary epithelium and demonstrates, for the first time a protective role for Dectin-1 blockade in epithelial defences. Infecting ΔpacC mutants are hypersensitive to cell wall-active antifungal agents highlighting the value of PacC signalling as a target for antifungal therapy.Public Library of Science (PLOS)Medical Research Council (Reino Unido)Biotechnology and Biological Sciences Research Council (Reino Unido)Wellcome TrustImperial College London (Reino Unido)FWF Austrian Science FundNational Institutes of Health (Estados Unidos)20182018-12-1320142014-10-1620142014-10-16journal articlehttp://purl.org/coar/resource_type/c_6501VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/20.500.12105/6837reponame:Repisaludinstname:Instituto de Salud Carlos III (ISCIII)Inglésengopen accesshttp://purl.org/coar/access_right/c_abf2Atribución 4.0 Internacionalhttp://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:repisalud.isciii.es:20.500.12105/68372026-06-12T12:43:37Z
dc.title.none.fl_str_mv The pH-responsive PacC transcription factor of Aspergillus fumigatus governs epithelial entry and tissue invasion during pulmonary aspergillosis
title The pH-responsive PacC transcription factor of Aspergillus fumigatus governs epithelial entry and tissue invasion during pulmonary aspergillosis
spellingShingle The pH-responsive PacC transcription factor of Aspergillus fumigatus governs epithelial entry and tissue invasion during pulmonary aspergillosis
Bertuzzi, Margherita
Animals
Aspergillus fumigatus
Epithelial Cells
Fungal Proteins
Hydrogen-Ion Concentration
Mice
Pulmonary Aspergillosis
Transcription Factors
title_short The pH-responsive PacC transcription factor of Aspergillus fumigatus governs epithelial entry and tissue invasion during pulmonary aspergillosis
title_full The pH-responsive PacC transcription factor of Aspergillus fumigatus governs epithelial entry and tissue invasion during pulmonary aspergillosis
title_fullStr The pH-responsive PacC transcription factor of Aspergillus fumigatus governs epithelial entry and tissue invasion during pulmonary aspergillosis
title_full_unstemmed The pH-responsive PacC transcription factor of Aspergillus fumigatus governs epithelial entry and tissue invasion during pulmonary aspergillosis
title_sort The pH-responsive PacC transcription factor of Aspergillus fumigatus governs epithelial entry and tissue invasion during pulmonary aspergillosis
dc.creator.none.fl_str_mv Bertuzzi, Margherita
Schrettl, Markus
Alcazar-Fuoli, Laura
Cairns, Timothy C
Muñoz, Alberto
Walker, Louise A
Herbst, Susanne
Safari, Maryam
Cheverton, Angela M
Chen, Dan
Liu, Hong
Saijo, Shinobu
Fedorova, Natalie D
Armstrong-James, Darius
Munro, Carol A
Read, Nick D
Filler, Scott G
Espeso, Eduardo A
Nierman, William C
Haas, Hubertus
Bignell, Elaine M
author Bertuzzi, Margherita
author_facet Bertuzzi, Margherita
Schrettl, Markus
Alcazar-Fuoli, Laura
Cairns, Timothy C
Muñoz, Alberto
Walker, Louise A
Herbst, Susanne
Safari, Maryam
Cheverton, Angela M
Chen, Dan
Liu, Hong
Saijo, Shinobu
Fedorova, Natalie D
Armstrong-James, Darius
Munro, Carol A
Read, Nick D
Filler, Scott G
Espeso, Eduardo A
Nierman, William C
Haas, Hubertus
Bignell, Elaine M
author_role author
author2 Schrettl, Markus
Alcazar-Fuoli, Laura
Cairns, Timothy C
Muñoz, Alberto
Walker, Louise A
Herbst, Susanne
Safari, Maryam
Cheverton, Angela M
Chen, Dan
Liu, Hong
Saijo, Shinobu
Fedorova, Natalie D
Armstrong-James, Darius
Munro, Carol A
Read, Nick D
Filler, Scott G
Espeso, Eduardo A
Nierman, William C
Haas, Hubertus
Bignell, Elaine M
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Medical Research Council (Reino Unido)
Biotechnology and Biological Sciences Research Council (Reino Unido)
Wellcome Trust
Imperial College London (Reino Unido)
FWF Austrian Science Fund
National Institutes of Health (Estados Unidos)

dc.subject.none.fl_str_mv Animals
Aspergillus fumigatus
Epithelial Cells
Fungal Proteins
Hydrogen-Ion Concentration
Mice
Pulmonary Aspergillosis
Transcription Factors
topic Animals
Aspergillus fumigatus
Epithelial Cells
Fungal Proteins
Hydrogen-Ion Concentration
Mice
Pulmonary Aspergillosis
Transcription Factors
description Destruction of the pulmonary epithelium is a major feature of lung diseases caused by the mould pathogen Aspergillus fumigatus. Although it is widely postulated that tissue invasion is governed by fungal proteases, A. fumigatus mutants lacking individual or multiple enzymes remain fully invasive, suggesting a concomitant requirement for other pathogenic activities during host invasion. In this study we discovered, and exploited, a novel, tissue non-invasive, phenotype in A. fumigatus mutants lacking the pH-responsive transcription factor PacC. Our study revealed a novel mode of epithelial entry, occurring in a cell wall-dependent manner prior to protease production, and via the Dectin-1 β-glucan receptor. ΔpacC mutants are defective in both contact-mediated epithelial entry and protease expression, and significantly attenuated for pathogenicity in leukopenic mice. We combined murine infection modelling, in vivo transcriptomics, and in vitro infections of human alveolar epithelia, to delineate two major, and sequentially acting, PacC-dependent processes impacting epithelial integrity in vitro and tissue invasion in the whole animal. We demonstrate that A. fumigatus spores and germlings are internalised by epithelial cells in a contact-, actin-, cell wall- and Dectin-1 dependent manner and ΔpacC mutants, which aberrantly remodel the cell wall during germinative growth, are unable to gain entry into epithelial cells, both in vitro and in vivo. We further show that PacC acts as a global transcriptional regulator of secreted molecules during growth in the leukopenic mammalian lung, and profile the full cohort of secreted gene products expressed during invasive infection. Our study reveals a combinatorial mode of tissue entry dependent upon sequential, and mechanistically distinct, perturbations of the pulmonary epithelium and demonstrates, for the first time a protective role for Dectin-1 blockade in epithelial defences. Infecting ΔpacC mutants are hypersensitive to cell wall-active antifungal agents highlighting the value of PacC signalling as a target for antifungal therapy.
publishDate 2014
dc.date.none.fl_str_mv 2014
2014-10-16
2014
2014-10-16
2018
2018-12-13
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
VoR
http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/20.500.12105/6837
url http://hdl.handle.net/20.500.12105/6837
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Atribución 4.0 Internacional
http://creativecommons.org/licenses/by/4.0/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
Atribución 4.0 Internacional
http://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Public Library of Science (PLOS)
publisher.none.fl_str_mv Public Library of Science (PLOS)
dc.source.none.fl_str_mv reponame:Repisalud
instname:Instituto de Salud Carlos III (ISCIII)
instname_str Instituto de Salud Carlos III (ISCIII)
reponame_str Repisalud
collection Repisalud
repository.name.fl_str_mv
repository.mail.fl_str_mv
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