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

被引：5
作者：
Calucho, Maite [1 ,2 ]
Gartner, Silvia [3 ]
Barranco, Paula [1 ,2 ]
Fernandez-Alvarez, Paula [1 ,2 ]
Perez, Raquel Garcia [4 ]
Tizzano, Eduardo F. [1 ,2 ]
机构：
[1] Vall DHebron Res Inst, Med Genet Grp, Barcelona 08035, Spain
[2] Hosp Univ Vall dHebron, Dept Clin & Mol Genet, Barcelona 08035, Spain
[3] Hosp Univ Vall dHebron, Cyst Fibrosis Unit, Barcelona 08035, Spain
[4] Autonomous Univ Barcelona, Bellaterra 08193, Spain
来源：
SCIENTIFIC REPORTS | 2021年 / 11卷 / 01期
关键词：
TEZACAFTOR-IVACAFTOR; IN-VITRO; LUMACAFTOR; POTENTIATOR; MUTATION; PROGRESS; THERAPY;
D O I：
10.1038/s41598-021-94798-x
中图分类号：
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号：
07 ; 0710 ; 09 ;
摘要：
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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页数：11
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