title: Activation of the cystic fibrosis transmembrane conductance regulator by the flavonoid quercetin: potential use as a biomarker of ΔF508 cystic fibrosis transmembrane conductance regulator rescue.
abstract: Therapies to correct the ΔF508 cystic fibrosis transmembrane conductance regulator (CFTR) folding defect require sensitive methods to detect channel activity in vivo. The β₂ adrenergic receptor agonists, which provide the CFTR stimuli commonly used in nasal potential difference assays, may not overcome the channel gating defects seen in ΔF508 CFTR after plasma membrane localization. In this study, we identify an agent, quercetin, that enhances the detection of surface ΔF508 CFTR, and is suitable for nasal perfusion. A screen of flavonoids in CFBE41o⁻ cells stably transduced with ΔF508 CFTR, corrected to the cell surface with low temperature growth, revealed that quercetin stimulated an increase in the short-circuit current. This increase was dose-dependent in both Fisher rat thyroid and CFBE41o⁻ cells. High concentrations inhibited Cl⁻ conductance. In CFBE41o⁻ airway cells, quercetin (20 μg/ml) activated ΔF508 CFTR, whereas the β₂ adrenergic receptor agonist isoproterenol did not. Quercetin had limited effects on cAMP levels, but did not produce detectable phosphorylation of the isolated CFTR R-domain, suggesting an activation independent of channel phosphorylation. When perfused in the nares of Cftr(+) mice, quercetin (20 μg/ml) produced a hyperpolarization of the potential difference that was absent in Cftr(-/-) mice. Finally, quercetin-induced, dose-dependent hyperpolarization of the nasal potential difference was also seen in normal human subjects. Quercetin activates CFTR-mediated anion transport in respiratory epithelia in vitro and in vivo, and may be useful in studies intended to detect the rescue of ΔF508 CFTR by nasal potential difference.
authors.affiliation: Department of Medicine, University of Alabama at Birmingham, Birmingham, Alabama, USA.
authors.forename: Louise C
authors.initials: LC
authors.lastname: Pyle
authors.forename: Jennifer C
authors.initials: JC
authors.lastname: Fulton
authors.forename: Peter A
authors.initials: PA
authors.lastname: Sloane
authors.forename: Kyle
authors.initials: K
authors.lastname: Backer
authors.forename: Marina
authors.initials: M
authors.lastname: Mazur
authors.forename: Jeevan
authors.initials: J
authors.lastname: Prasain
authors.forename: Stephen
authors.initials: S
authors.lastname: Barnes
authors.forename: J P
authors.initials: JP
authors.lastname: Clancy
authors.forename: Steven M
authors.initials: SM
authors.lastname: Rowe
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002712:Chlorides / Q000378:metabolism; D000242:Cyclic AMP / Q000378:metabolism; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000737:chemistry / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical; D006801:Humans; D015640:Ion Channel Gating / Q000187:drug effects*; D017136:Ion Transport / Q000187:drug effects; D008564:Membrane Potentials / Q000187:drug effects; D051379:Mice; D050505:Mutant Proteins / Q000737:chemistry / Q000378:metabolism*; D041681:NIH 3T3 Cells; D009297:Nasal Mucosa / Q000187:drug effects / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D017434:Protein Structure, Tertiary; D011794:Quercetin / Q000494:pharmacology*; D051381:Rats; D018343:Receptors, Adrenergic, beta-2 / Q000378:metabolism
chemical_list: D015415:Biomarkers; D002712:Chlorides; D050505:Mutant Proteins; D018343:Receptors, Adrenergic, beta-2; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D011794:Quercetin; D000242:Cyclic AMP