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.
Highlights
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
We first validated the formation of WT nasospheroids in cell cultures from epithelial nasal cells obtained by nasal curettage
CFTR FSK/cAMP-stimulated transport was studied in nasospheroids by live-cell imaging to analyze shrinking after CFTR activation 30
Summary
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. We implemented the measurement of inner fluid reservoir areas to further differentiate CFTR functionality This rapid methodology is helpful to analyse response to modulators in CF samples to allow individualized treatment for CF patients. Cystic fibrosis (CF) is the most common lethal monogenic disease with an incidence of 1/3000–10,000 newborns It is caused by bi-allelic pathogenic variants in the cystic fibrosis transmembrane regulator (CFTR) gene and codifies for an epithelial Cl− channel 1. A triple combination with two different correctors (tezacaftor and elexacaftor) and potentiator ivacaftor (TRIKAFTA/KAFTRIO), has been approved to treat F508del homozygous patients [10,14], or those who carry a single copy of the F508del genetic variant with a minimal function variant [15,16]. It is crucial to study CFTR function individually in order to predict responses to currently approved or experimental drugs
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