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Validation of nasospheroids to assay CFTR functionality and modulator responses in cystic fibrosis

The availability of a simple, robust and non-invasive in vitro airway model would be useful to study the functionality of the cystic fibrosis transmembrane regulator (CFTR) protein and to personalize modulator therapy for cystic fibrosis (CF) patients. Our aim was to validate a CFTR functional study...

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Autores principales: Calucho, Maite, Gartner, Silvia, Barranco, Paula, Fernández-Álvarez, Paula, Pérez, Raquel García, Tizzano, Eduardo F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8324871/
https://www.ncbi.nlm.nih.gov/pubmed/34330959
http://dx.doi.org/10.1038/s41598-021-94798-x
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author Calucho, Maite
Gartner, Silvia
Barranco, Paula
Fernández-Álvarez, Paula
Pérez, Raquel García
Tizzano, Eduardo F.
author_facet Calucho, Maite
Gartner, Silvia
Barranco, Paula
Fernández-Álvarez, Paula
Pérez, Raquel García
Tizzano, Eduardo F.
author_sort Calucho, Maite
collection PubMed
description The availability of a simple, robust and non-invasive in vitro airway model would be useful to study the functionality of the cystic fibrosis transmembrane regulator (CFTR) protein and to personalize modulator therapy for cystic fibrosis (CF) patients. Our aim was to validate a CFTR functional study using nasospheroids, a patient-derived nasal cell 3D-culture. We performed live-cell experiments in nasospheroids obtained from wild-type individuals and CF patients with different genotypes and phenotypes. We extended the existing method and expanded the analysis to upgrade measurements of CFTR activity using forskolin-induced shrinking. We also tested modulator drugs in CF samples. Immobilizing suspended-nasospheroids provided a high number of samples for live-cell imaging. The diversity observed in basal sizes of nasospheroids did not affect the functional analysis of CFTR. Statistical analysis with our method was simple, making this protocol easy to reproduce. Moreover, we implemented the measurement of inner fluid reservoir areas to further differentiate CFTR functionality. In summary, this rapid methodology is helpful to analyse response to modulators in CF samples to allow individualized treatment for CF patients.
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spelling pubmed-83248712021-08-02 Validation of nasospheroids to assay CFTR functionality and modulator responses in cystic fibrosis Calucho, Maite Gartner, Silvia Barranco, Paula Fernández-Álvarez, Paula Pérez, Raquel García Tizzano, Eduardo F. Sci Rep Article The availability of a simple, robust and non-invasive in vitro airway model would be useful to study the functionality of the cystic fibrosis transmembrane regulator (CFTR) protein and to personalize modulator therapy for cystic fibrosis (CF) patients. Our aim was to validate a CFTR functional study using nasospheroids, a patient-derived nasal cell 3D-culture. We performed live-cell experiments in nasospheroids obtained from wild-type individuals and CF patients with different genotypes and phenotypes. We extended the existing method and expanded the analysis to upgrade measurements of CFTR activity using forskolin-induced shrinking. We also tested modulator drugs in CF samples. Immobilizing suspended-nasospheroids provided a high number of samples for live-cell imaging. The diversity observed in basal sizes of nasospheroids did not affect the functional analysis of CFTR. Statistical analysis with our method was simple, making this protocol easy to reproduce. Moreover, we implemented the measurement of inner fluid reservoir areas to further differentiate CFTR functionality. In summary, this rapid methodology is helpful to analyse response to modulators in CF samples to allow individualized treatment for CF patients. Nature Publishing Group UK 2021-07-30 /pmc/articles/PMC8324871/ /pubmed/34330959 http://dx.doi.org/10.1038/s41598-021-94798-x Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Calucho, Maite
Gartner, Silvia
Barranco, Paula
Fernández-Álvarez, Paula
Pérez, Raquel García
Tizzano, Eduardo F.
Validation of nasospheroids to assay CFTR functionality and modulator responses in cystic fibrosis
title Validation of nasospheroids to assay CFTR functionality and modulator responses in cystic fibrosis
title_full Validation of nasospheroids to assay CFTR functionality and modulator responses in cystic fibrosis
title_fullStr Validation of nasospheroids to assay CFTR functionality and modulator responses in cystic fibrosis
title_full_unstemmed Validation of nasospheroids to assay CFTR functionality and modulator responses in cystic fibrosis
title_short Validation of nasospheroids to assay CFTR functionality and modulator responses in cystic fibrosis
title_sort validation of nasospheroids to assay cftr functionality and modulator responses in cystic fibrosis
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8324871/
https://www.ncbi.nlm.nih.gov/pubmed/34330959
http://dx.doi.org/10.1038/s41598-021-94798-x
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