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cohort·pulmonology, pediatrics, research methods, clinical trial, preclinical research·PMC10136647

Correlation of Electrophysiological and Fluorescence-Based Measurements of Modulator Efficacy in Nasal Epithelial Cultures Derived from People with Cystic Fibrosis

Cells · 9 authors, 7 centres

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TL;DR

The study compared two laboratory methods for measuring the effectiveness of CFTR modulator drugs in nasal cell cultures from cystic fibrosis patients. The faster fluorescence-based assay produced results that correlated with the standard, slower electrophysiology method for cultures with the F508del mutation and showed potential for detecting responses in cultures with the W1282X mutation.

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This methodological study compared two in vitro assays for measuring CFTR modulator responses in primary nasal epithelial cultures from 39 cystic fibrosis patients with different genotypes. The researchers used both the standard Ussing chamber technique and a fluorescence-based, multi-transwell assay called Fl-ACC or FMP. The central result was that the fluorescence assay successfully detected positive responses across all tested genotypes and its results for F508del cultures correlated with the Ussing chamber findings. The fluorescence assay also showed a trend of greater sensitivity for detecting responses in cultures with the W1282X mutation. The findings support the use of the faster fluorescence method as a complementary or alternative tool for theratyping to guide potential therapies for rare mutations.

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cohort

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pulmonology

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ORIGINAL PAPER

PMC10136647

Correlation of Electrophysiological and Fluorescence-Based Measurements of Modulator Efficacy in Nasal Epithelial Cultures Derived from People with Cystic Fibrosis

Cells

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ABSTRACT

It has been suggested that in vitro studies of the rescue effect of CFTR modulator drugs in nasal epithelial cultures derived from people with cystic fibrosis have the potential to predict clinical responses to the same drugs. Hence, there is an interest in evaluating different methods for measuring in vitro modulator responses in patient-derived nasal cultures. Commonly, the functional response to CFTR modulator combinations in these cultures is assessed by bioelectric measurements, using the Ussing chamber. While this method is highly informative, it is time-consuming. A fluorescence-based, multi-transwell method for assaying regulated apical chloride conductance (Fl-ACC) promises to provide a complementary approach to theratyping in patient-derived nasal cultures. In the present work, we compared Ussing chamber measurements and fluorescence-based measurements of CFTR-mediated apical conductance in matching, fully differentiated nasal cultures derived from CF patients, homozygous for F508del ( n = 31) or W1282X ( n = 3), or heterozygous for Class III mutations G551D or G178R ( n = 5). These cultures were obtained through a bioresource called the Cystic Fibrosis Canada-Sick Kids Program in Individual CF Therapy (CFIT). We found that the Fl-ACC method was effective in detecting positive responses to interventions for all genotypes. There was a correlation between patient-specific drug responses measured in cultures harbouring F508del, as measured using the Ussing chamber technique and the fluorescence-based assay (Fl-ACC). Finally, the fluorescence-based assay has the potential for greater sensitivity for detecting responses to pharmacological rescue strategies targeting W1282X.

CITATION EXPORT

Tarini N. A. Gunawardena, Zoltán Bozóky, Claire Bartlett et al.. Correlation of Electrophysiological and Fluorescence-Based Measurements of Modulator Efficacy in Nasal Epithelial Cultures Derived from People with Cystic Fibrosis. Cells. (2023). https://doi.org/10.3390/cells12081174

AUTHORS

9 · 7 CENTRES

Tarini N. A. Gunawardena, Zoltán Bozóky, Claire Bartlett, Hong Ouyang, Paul D. W. Eckford, Theo J. Moraes, Felix Ratjen, Tanja Gonska, Christine E. Bear

1Program of Molecular Medicine, The Hospital for Sick Children, Toronto, ON M5G 1X8, Canada
2Program of Translational Medicine, The Hospital for Sick Children, Toronto, ON M5G 1X8, Canada
3Providence Health Care, Vancouver, BC V6Z 1Y6, Canada
4Department of Paediatrics, Division of Respiratory Medicine, The Hospital for Sick Children, Toronto, ON M5G 1X8, Canada
5Department of Paediatrics, Division of Gastroenterology, Hepatology and Nutrition, University of Toronto, Toronto, ON M5S 1A8, Canada
6Department of Physiology, University of Toronto, Toronto, ON M5S 1A8, Canada
7Department of Biochemistry, University of Toronto, Toronto, ON M5S 1A8, Canada

EDITOR / REVIEWER

Elio Castagnola