Measurements of spontaneous CFTR-mediated ion transport without acute channel activation in airway epithelial cultures after modulator exposure

• Published in: Scientific reports Vol. 11; no. 1; p. 22616
• Main Authors: Nick, Heidi J, Zeitlin, Pamela L, Yadav, Sangya, Bratcher, Preston E
• Format: Journal Article
• Language: English
• Published: England Nature Publishing Group 19.11.2021; Nature Publishing Group UK; Nature Portfolio
• Subjects: 1-Methyl-3-isobutylxanthine - pharmacology; Amiloride; Aminophenols; Aminopyridines - pharmacology; Animals; Benzodioxoles - pharmacology; Biological Products; Cells, Cultured; Colforsin - pharmacology; Cystic Fibrosis - metabolism; Cystic Fibrosis - therapy; Cystic Fibrosis Transmembrane Conductance Regulator - chemistry; Cystic Fibrosis Transmembrane Conductance Regulator - genetics; Electrophysiology - methods; Epithelial Cells - metabolism; Epithelium - metabolism; Forskolin; Genotype; Humans; Ion transport; Ions; Mice; NIH 3T3 Cells; Quantitation; Quinolones; Respiratory tract
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-021-02044-1

Quantitation of CFTR function in vitro is commonly performed by acutely stimulating then inhibiting ion transport through CFTR and measuring the resulting changes in transepithelial voltage (V ) and current (I ). While this technique is suitable for measuring the maximum functional capacity of CFTR, it may not provide an accurate estimate of in vivo CFTR activity. To test if CFTR-mediated ion transport could be measured in the absence of acute CFTR stimulation, primary airway epithelia were analyzed in an Ussing chamber with treatment of amiloride followed by CFTR(inh)-172 without acute activation of CFTR. Non-CF epithelia demonstrated a decrease in V and I following exposure to CFTR(inh)-172 and in the absence of forskolin/IBMX (F/I); this decrease is interpreted as a measure of spontaneous CFTR activity present in these epithelia. In F508del/F508del CFTR epithelia, F/I-induced changes in V and I were ~ fourfold increased after treatment with VX-809/VX-770, while the magnitude of spontaneous CFTR activities were only ~ 1.6-fold increased after VX-809/VX-770 treatment. Method-dependent discrepancies in the responses of other CF epithelia to modulator treatments were observed. These results serve as a proof of concept for the analysis of CFTR modulator responses in vitro in the absence of acute CFTR activation. Future studies will determine the usefulness of this approach in the development of novel CFTR modulator therapies.