Journal Article

Modulation of deltaF508 cystic fibrosis transmembrane regulator trafficking and function with 4-phenylbutyrate and flavonoids

Floyd, Alexandra D, Kwon, Edwin, McKenzie, Karen, Lim, Meerana, Zeitlin, Pamela L

2004

Overview

Over 70% of patients with cystic fibrosis have the DeltaF508 mutation. This protein is a partially functional chloride (Cl-) channel that is prematurely degraded in the endoplasmic reticulum. Specific members of the flavonoid class of compounds have been shown to increase Cl- conductance of wild-type and DeltaF508 cystic fibrosis transmembrane regulator (CFTR). Although flavonoid effects on CFTR processing are unknown, evidence of effects on heat shock proteins, specifically those that have been shown to interact with CFTR, led us to believe that there would be an effect on CFTR processing through modulation of CFTR-chaperone interactions. We sought to determine (i) the effect of apigenin, genistein, kaempferol, and quercetin on CFTR processing in IB3-1 cells (F508/W1282X) and (ii) whether sequential treatment with 4-phenylbutyrate (4-PBA) to increase CFTR processing and flavonoid to directly stimulate CFTR would increase Cl- conductance. Our results show no significant effect on CFTR processing as measured by immunoblotting with 1 microM or 5 microM of apigenin, genistein, kaempferol, or quercetin. However, despite no effect on CFTR processing as determined by immunoblot, immunofluorescence demonstrated a favorable change in the intracellular distribution of CFTR with 24 h treatments of apigenin, kaempferol, and genistein. Furthermore, we observed an increase in Cl- conductance as measured by Cl- efflux in cells that were treated for 24 h with 4-PBA and then assayed with forskolin and 1 microM or 5 microM genistein, and also with cells treated for 24 h with either 4-PBA, 5 microM apigenin, or 1 microM quercetin. Thus, a combination of chronic treatment with 4-PBA or select flavonoids, followed by acute flavonoid exposure, may be beneficial in cystic fibrosis.

• Subject: Chlorides - metabolism / Cystic Fibrosis - genetics / Cystic Fibrosis Transmembrane Conductance Regulator - drug effects / Cystic Fibrosis Transmembrane Conductance Regulator - genetics / Cystic Fibrosis Transmembrane Conductance Regulator - metabolism / Dose-Response Relationship, Drug / Drug Therapy, Combination / Flavonoids - pharmacology / Flavonoids - therapeutic use / Humans / Molecular Chaperones - drug effects / Molecular Chaperones - metabolism / Phenylbutyrates - pharmacology / Phenylbutyrates - therapeutic use / Protein Folding / Protein Transport - drug effects / Protein Transport - physiology / Respiratory Mucosa - drug effects / Respiratory Mucosa - metabolism / Respiratory Mucosa - physiopathology / Up-Regulation - drug effects / Up-Regulation - physiology