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CFTR Rescue by Lumacaftor (VX-809) Induces an Extensive Reorganization of Mitochondria in the Cystic Fibrosis Bronchial Epithelium
Background: Cystic Fibrosis (CF) is a genetic disorder affecting around 1 in every 3000 newborns. In the most common mutation, F508del, the defective anion channel, CFTR, is prevented from reaching the plasma membrane (PM) by the quality check control of the cell. Little is known about how CFTR phar...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9222197/ https://www.ncbi.nlm.nih.gov/pubmed/35741067 http://dx.doi.org/10.3390/cells11121938 |
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author | Braccia, Clarissa Christopher, Josie A. Crook, Oliver M. Breckels, Lisa M. Queiroz, Rayner M. L. Liessi, Nara Tomati, Valeria Capurro, Valeria Bandiera, Tiziano Baldassari, Simona Pedemonte, Nicoletta Lilley, Kathryn S. Armirotti, Andrea |
author_facet | Braccia, Clarissa Christopher, Josie A. Crook, Oliver M. Breckels, Lisa M. Queiroz, Rayner M. L. Liessi, Nara Tomati, Valeria Capurro, Valeria Bandiera, Tiziano Baldassari, Simona Pedemonte, Nicoletta Lilley, Kathryn S. Armirotti, Andrea |
author_sort | Braccia, Clarissa |
collection | PubMed |
description | Background: Cystic Fibrosis (CF) is a genetic disorder affecting around 1 in every 3000 newborns. In the most common mutation, F508del, the defective anion channel, CFTR, is prevented from reaching the plasma membrane (PM) by the quality check control of the cell. Little is known about how CFTR pharmacological rescue impacts the cell proteome. Methods: We used high-resolution mass spectrometry, differential ultracentrifugation, machine learning and bioinformatics to investigate both changes in the expression and localization of the human bronchial epithelium CF model (F508del-CFTR CFBE41o-) proteome following treatment with VX-809 (Lumacaftor), a drug able to improve the trafficking of CFTR. Results: The data suggested no stark changes in protein expression, yet subtle localization changes of proteins of the mitochondria and peroxisomes were detected. We then used high-content confocal microscopy to further investigate the morphological and compositional changes of peroxisomes and mitochondria under these conditions, as well as in patient-derived primary cells. We profiled several thousand proteins and we determined the subcellular localization data for around 5000 of them using the LOPIT-DC spatial proteomics protocol. Conclusions: We observed that treatment with VX-809 induces extensive structural and functional remodelling of mitochondria and peroxisomes that resemble the phenotype of healthy cells. Our data suggest additional rescue mechanisms of VX-809 beyond the correction of aberrant folding of F508del-CFTR and subsequent trafficking to the PM. |
format | Online Article Text |
id | pubmed-9222197 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-92221972022-06-24 CFTR Rescue by Lumacaftor (VX-809) Induces an Extensive Reorganization of Mitochondria in the Cystic Fibrosis Bronchial Epithelium Braccia, Clarissa Christopher, Josie A. Crook, Oliver M. Breckels, Lisa M. Queiroz, Rayner M. L. Liessi, Nara Tomati, Valeria Capurro, Valeria Bandiera, Tiziano Baldassari, Simona Pedemonte, Nicoletta Lilley, Kathryn S. Armirotti, Andrea Cells Article Background: Cystic Fibrosis (CF) is a genetic disorder affecting around 1 in every 3000 newborns. In the most common mutation, F508del, the defective anion channel, CFTR, is prevented from reaching the plasma membrane (PM) by the quality check control of the cell. Little is known about how CFTR pharmacological rescue impacts the cell proteome. Methods: We used high-resolution mass spectrometry, differential ultracentrifugation, machine learning and bioinformatics to investigate both changes in the expression and localization of the human bronchial epithelium CF model (F508del-CFTR CFBE41o-) proteome following treatment with VX-809 (Lumacaftor), a drug able to improve the trafficking of CFTR. Results: The data suggested no stark changes in protein expression, yet subtle localization changes of proteins of the mitochondria and peroxisomes were detected. We then used high-content confocal microscopy to further investigate the morphological and compositional changes of peroxisomes and mitochondria under these conditions, as well as in patient-derived primary cells. We profiled several thousand proteins and we determined the subcellular localization data for around 5000 of them using the LOPIT-DC spatial proteomics protocol. Conclusions: We observed that treatment with VX-809 induces extensive structural and functional remodelling of mitochondria and peroxisomes that resemble the phenotype of healthy cells. Our data suggest additional rescue mechanisms of VX-809 beyond the correction of aberrant folding of F508del-CFTR and subsequent trafficking to the PM. MDPI 2022-06-16 /pmc/articles/PMC9222197/ /pubmed/35741067 http://dx.doi.org/10.3390/cells11121938 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Braccia, Clarissa Christopher, Josie A. Crook, Oliver M. Breckels, Lisa M. Queiroz, Rayner M. L. Liessi, Nara Tomati, Valeria Capurro, Valeria Bandiera, Tiziano Baldassari, Simona Pedemonte, Nicoletta Lilley, Kathryn S. Armirotti, Andrea CFTR Rescue by Lumacaftor (VX-809) Induces an Extensive Reorganization of Mitochondria in the Cystic Fibrosis Bronchial Epithelium |
title | CFTR Rescue by Lumacaftor (VX-809) Induces an Extensive Reorganization of Mitochondria in the Cystic Fibrosis Bronchial Epithelium |
title_full | CFTR Rescue by Lumacaftor (VX-809) Induces an Extensive Reorganization of Mitochondria in the Cystic Fibrosis Bronchial Epithelium |
title_fullStr | CFTR Rescue by Lumacaftor (VX-809) Induces an Extensive Reorganization of Mitochondria in the Cystic Fibrosis Bronchial Epithelium |
title_full_unstemmed | CFTR Rescue by Lumacaftor (VX-809) Induces an Extensive Reorganization of Mitochondria in the Cystic Fibrosis Bronchial Epithelium |
title_short | CFTR Rescue by Lumacaftor (VX-809) Induces an Extensive Reorganization of Mitochondria in the Cystic Fibrosis Bronchial Epithelium |
title_sort | cftr rescue by lumacaftor (vx-809) induces an extensive reorganization of mitochondria in the cystic fibrosis bronchial epithelium |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9222197/ https://www.ncbi.nlm.nih.gov/pubmed/35741067 http://dx.doi.org/10.3390/cells11121938 |
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