title: An "ex vivo model" contributing to the diagnosis and evaluation of new drugs in cystic fibrosis.
abstract: Cystic fibrosis (CF) is an autosomal recessive disease caused by mutations in the cystic fibrosis transmembrane regulator (CFTR) gene. About 2000 mutations have been described so far. We setup an ex vivo model of human nasal epithelial cells (HNECs) to study CF patients testing the effect of novel mutations and molecular therapies. We performed sampling (by brushing), followed by culture and analysis of HNECs using a series of molecular techniques. We performed 50 brushings from CF patients and controls. Using cultured cells, we: i) demonstrated the widely heterogeneous CFTR expression in patients and in controls; ii) defined the splicing effect of a CFTR mutation; iii) assessed the CFTR gating activity in patients bearing different mutations; iv) demonstrated that butyrate significantly enhances CFTR expression. Based on our data, we can conclude: 1) HNEC brushing is performed without anaesthesia and is well tolerated in all CF patients (children and adults); 2) HNECs can be preserved for up to 48 hours before culture allowings multicentre studies; 3) HNECs culture can be considered a suitable model to study the molecular effects of new CFTR gene mutations and/or uncertain meaning specific mutations of carriers; 4) an ex vivo model of HNECs may be used to evaluate, before human use, the effect of new drugs on patients' cells bearing specific CFTR mutations; 5) the methodology is adequate for a quantitative measurement, by fluorescence, of the CFTR gating activity of the HNECs from patients with different genotypes identifying: a) CF patients bearing two severe mutations with an activity < 10% (compared to controls - 100%); b) CF patients bearing at least a mild mutation with an activity of 10-20%; c) CF carriers (heterozygous subjects) with an activity between 40-70%.
authors.affiliation: Department of Neuroscience, Section of Otorhinolaryngology, University of Naples "Federico II", Italy.|CEINGE-Advanced Biotechnologies scarl, Naples, Italy.|Department of Molecular Medicine and Medical Biotechnologies, University of Naples "Federico II", Italy.
authors.forename: A M
authors.initials: AM
authors.lastname: Di Lullo
authors.affiliation: CEINGE-Advanced Biotechnologies scarl, Naples, Italy.|Department of Molecular Medicine and Medical Biotechnologies, University of Naples "Federico II", Italy.
authors.forename: M
authors.initials: M
authors.lastname: Scorza
authors.affiliation: CEINGE-Advanced Biotechnologies scarl, Naples, Italy.|Department of Molecular Medicine and Medical Biotechnologies, University of Naples "Federico II", Italy.
authors.forename: F
authors.initials: F
authors.lastname: Amato
authors.affiliation: CEINGE-Advanced Biotechnologies scarl, Naples, Italy.|Department of Molecular Medicine and Medical Biotechnologies, University of Naples "Federico II", Italy.
authors.forename: M
authors.initials: M
authors.lastname: Comegna
authors.affiliation: Department of Translational Medical Sciences, University of Naples "Federico II", Italy.
authors.forename: V
authors.initials: V
authors.lastname: Raia
authors.affiliation: Department of Science and Technology Innovation, University of Piemonte Orientale, Novara, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Maiuri
authors.affiliation: European Institute for Research in Cystic Fibrosis, San Raffaele Hospital, Milan, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Ilardi
authors.affiliation: Department of Neuroscience, Section of Otorhinolaryngology, University of Naples "Federico II", Italy.
authors.forename: E
authors.initials: E
authors.lastname: Cantone
authors.affiliation: CEINGE-Advanced Biotechnologies scarl, Naples, Italy.|Department of Molecular Medicine and Medical Biotechnologies, University of Naples "Federico II", Italy.
authors.forename: G
authors.initials: G
authors.lastname: Castaldo
authors.affiliation: Department of Neuroscience, Section of Otorhinolaryngology, University of Naples "Federico II", Italy.
authors.forename: M
authors.initials: M
authors.lastname: Iengo
mesh_terms: D002478:Cells, Cultured; D003550:Cystic Fibrosis / Q000188:drug therapy* / Q000235:genetics; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000032:analysis / Q000235:genetics; D006801:Humans; D009154:Mutation; D009297:Nasal Mucosa / Q000166:cytology
chemical_list: D019005:Cystic Fibrosis Transmembrane Conductance Regulator
keywords: CF; CFTR; Cystic Fibrosis; Mutations; Nasal brushing