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

Abstract 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 functio...

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Autores principales:	Maite Calucho, Silvia Gartner, Paula Barranco, Paula Fernández-Álvarez, Raquel García Pérez, Eduardo F. Tizzano
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/ceebc254ff6749ddae89f16c9505710e
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spelling	oai:doaj.org-article:ceebc254ff6749ddae89f16c9505710e2021-12-02T18:47:07ZValidation of nasospheroids to assay CFTR functionality and modulator responses in cystic fibrosis10.1038/s41598-021-94798-x2045-2322https://doaj.org/article/ceebc254ff6749ddae89f16c9505710e2021-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-94798-xhttps://doaj.org/toc/2045-2322Abstract 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.Maite CaluchoSilvia GartnerPaula BarrancoPaula Fernández-ÁlvarezRaquel García PérezEduardo F. TizzanoNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-11 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Maite Calucho Silvia Gartner Paula Barranco Paula Fernández-Álvarez Raquel García Pérez Eduardo F. Tizzano Validation of nasospheroids to assay CFTR functionality and modulator responses in cystic fibrosis
description	Abstract 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.
format	article
author	Maite Calucho Silvia Gartner Paula Barranco Paula Fernández-Álvarez Raquel García Pérez Eduardo F. Tizzano
author_facet	Maite Calucho Silvia Gartner Paula Barranco Paula Fernández-Álvarez Raquel García Pérez Eduardo F. Tizzano
author_sort	Maite Calucho
title	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_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_sort	validation of nasospheroids to assay cftr functionality and modulator responses in cystic fibrosis
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/ceebc254ff6749ddae89f16c9505710e
work_keys_str_mv	AT maitecalucho validationofnasospheroidstoassaycftrfunctionalityandmodulatorresponsesincysticfibrosis AT silviagartner validationofnasospheroidstoassaycftrfunctionalityandmodulatorresponsesincysticfibrosis AT paulabarranco validationofnasospheroidstoassaycftrfunctionalityandmodulatorresponsesincysticfibrosis AT paulafernandezalvarez validationofnasospheroidstoassaycftrfunctionalityandmodulatorresponsesincysticfibrosis AT raquelgarciaperez validationofnasospheroidstoassaycftrfunctionalityandmodulatorresponsesincysticfibrosis AT eduardoftizzano validationofnasospheroidstoassaycftrfunctionalityandmodulatorresponsesincysticfibrosis
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Validation of nasospheroids to assay CFTR functionality and modulator responses in cystic fibrosis

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