Alain Palem

41 Corrector effect of resveratrol on refractory adrenergic pathway in saliva secretion of cystic fibrosis mice

Objectives: Expression of the cystic fibrosis transmembrane conductance regulator is altered in individuals with the DF508 CFTR mutation. MicroRNA (miRNA) are differentially expressed in the CF airway epithelium, however their role in regulation of CFTR expression here remains unexplored. Herein we investigated the role of up regulated miRNAs in CFTR regulation in vivo in bronchial brushings from individuals homozygous or heterozygous for DF508 CFTR, validated our observations in vitro and assessed the impact of defective chloride ion conductance, genotype and Pseudomonas status on miRNA expression. Methods: MiRNA target prediction was performed in silico, expression of miRNAs and target genes was measured by qRT-PCR and/or western blotting. Modulation of miRNA was carried out using pre-miRs or antagomirs and a luciferase reporter gene employed to elucidate miRNA/CFTR interaction. MiR-145, -223 and 494 were up regulated in CF versus non-CF bronchial brushings and cell lines, in DF508 homovs. heterozygotes, in subjects positive for P. aeruginosa and in 16HBE14o− cells treated with a CFTR inhibitor. Reciprocal down or up regulation of CFTR gene and/or protein expression was observed following miRNA manipulation and direct miRNA/target relationship demonstrated via a luciferase reporter system containing the full length 3’-untranslated region of CFTR. Conclusions: Increased expression of miR-145, -223 and -494 in vivo in bronchial epithelium of individuals carrying the DF508 CFTR mutation correlates with decreased CFTR expression. Defective CFTR function and Pseudomonas colonisation may affect miRNA expression and contribute to the regulation of DF508 CFTR mutation.

63 Resveratrol improves chloride secretion in cystic fibrosis mice homozygous for the F508del mutation

63 Resveratrol improves chloride secretion in cystic fibrosis mice homozygous for the F508del mutation A. Palem1, C. Bouckaert1, B. Dhooghe1, A. Leonard2, B. Lubamba1, P. Wallemacq1, P. Lebecque2, T. Leal1. 1Universite Catholique de Louvain, Centre for Toxicology and Applied Pharmacology, Brussels, Belgium; 2Universite Catholique de Louvain, Pediatric Pulmonology & Cystic Fibrosis Unit, Cliniques Saint Luc, Brussels, Belgium