Electrophysiological evaluation of Cystic Fibrosis Conductance Transmembrane Regulator (CFTR) expression in human monocytes |
| |
Authors: | Michele Ettorre Genny Verzè Sara Caldrer Jan Johansson Elisa Calcaterra Baroukh Maurice Assael Paola Melotti Claudio Sorio Mario Buffelli |
| |
Institution: | 1. Department of Neurological and Movement Sciences, Section of Physiology, University of Verona, Strada Le Grazie 8, 37134 Verona, Italy;2. Department of Pathology and Diagnostics, University of Verona, Strada Le Grazie 8, 37134 Verona, Italy;3. Cystic Fibrosis Center, Azienda Ospedaliera Universitaria Integrata di Verona, Piazzale Stefani 1, 37126 Verona, Italy;4. Center for Biomedical Computing, University of Verona, Strada le Grazie 8, 37134 Verona, Italy;5. National Institute of Neuroscience, Verona, Italy |
| |
Abstract: | BackgroundCystic fibrosis is caused by mutations of CFTR gene, a protein kinase A-activated anion channel, and is associated to a persistent and excessive chronic lung inflammation, suggesting functional alterations of immune cells. Leukocytes express detectable levels of CFTR but the molecule has not been fully characterized in these cells.MethodsFreshly isolated monocytes from healthy individuals and CF patients were assessed by protein expression, single cell electrophysiological and membrane depolarization assays.ResultsWe recorded chloride currents by patch clamp in healthy monocytes, after the administration of a CFTR stimulus. Currents were sensitive to a specific blocker of the CFTR channel, CFTRinh-172 and were absent in CF monocytes. Next, we evaluated the effects of ex vivo exposure of monocytes from cystic fibrosis patients carrying the F508del mutation to a chemical corrector, Vertex-325. We found an increase in CFTR expression by confocal microscopy and a recovery of CFTR function by both patch clamp and single cell fluorescence analysis.ConclusionsWe confirm the expression of functional CFTR in human monocytes and demonstrate that blood monocytes can represent an adequate source of primary cells to assess new therapies and define diagnosis of CF.General significanceTests to evaluate CFTR functional abnormalities in CF disease might greatly benefit from the availability of a convenient source of primary cells. This electrophysiological study promotes the use of monocytes as a minimally invasive tool to study and monitor CFTR function in individual patients. |
| |
Keywords: | CF Cystic fibrosis CFTR Cystic Fibrosis Conductance Transmembrane Regulator CFTRinh-172 CFTR inhibitor F508del Deletion of phenylalanine at position 508 of the CFTR gene ICM Intestinal current measurement NPD Nasal potential difference S E Standard error Vertex-325 4-cyclohexyloxy-2-(1-[4-(4-methoxy-benzenesulfonyl)-piperazin-1-yl]-ethyl)-quinazoline WCR Whole cell recording |
本文献已被 ScienceDirect 等数据库收录! |
|