title: Peroxisome proliferator-activated receptor alpha, PPARα, directly regulates transcription of cytochrome P450 CYP2C8.
abstract: The cytochrome P450, CYP2C8, metabolizes more than 60 clinically used drugs as well as endogenous substances including retinoic acid and arachidonic acid. However, predictive factors for interindividual variability in the efficacy and toxicity of CYP2C8 drug substrates are essentially lacking. Recently we demonstrated that peroxisome proliferator-activated receptor alpha (PPARα), a nuclear receptor primarily involved in control of lipid and energy homeostasis directly regulates the transcription of CYP3A4. Here we investigated the potential regulation of CYP2C8 by PPARα. Two linked intronic SNPs in PPARα (rs4253728, rs4823613) previously associated with hepatic CYP3A4 status showed significant association with CYP2C8 protein level in human liver samples (N = 150). Furthermore, siRNA-mediated knock-down of PPARα in HepaRG human hepatocyte cells resulted in up to ∼60 and ∼50% downregulation of CYP2C8 mRNA and activity, while treatment with the PPARα agonist WY14,643 lead to an induction by >150 and >100%, respectively. Using chromatin immunoprecipitation scanning assay we identified a specific upstream gene region that is occupied in vivo by PPARα. Electromobility shift assay demonstrated direct binding of PPARα to a DR-1 motif located at positions -2762/-2775 bp upstream of the CYP2C8 transcription start site. We further validated the functional activity of this element using luciferase reporter gene assays in HuH7 cells. Moreover, based on our previous studies we demonstrated that WNT/β-catenin acts as a functional inhibitor of PPARα-mediated inducibility of CYP2C8 expression. In conclusion, our data suggest direct involvement of PPARα in both constitutive and inducible regulation of CYP2C8 expression in human liver, which is further modulated by WNT/β-catenin pathway. PPARA gene polymorphism could have a modest influence on CYP2C8 phenotype.
authors.affiliation: Dr. Margarete Fischer-Bosch-Institute of Clinical Pharmacology Stuttgart, Germany ; University of Tuebingen Tuebingen, Germany.
authors.forename: Maria
authors.initials: M
authors.lastname: Thomas
authors.affiliation: Dr. Margarete Fischer-Bosch-Institute of Clinical Pharmacology Stuttgart, Germany ; University of Tuebingen Tuebingen, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Winter
authors.affiliation: Dr. Margarete Fischer-Bosch-Institute of Clinical Pharmacology Stuttgart, Germany ; University of Tuebingen Tuebingen, Germany.
authors.forename: Britta
authors.initials: B
authors.lastname: Klumpp
authors.affiliation: Dr. Margarete Fischer-Bosch-Institute of Clinical Pharmacology Stuttgart, Germany ; University of Tuebingen Tuebingen, Germany.
authors.forename: Miia
authors.initials: M
authors.lastname: Turpeinen
authors.affiliation: Dr. Margarete Fischer-Bosch-Institute of Clinical Pharmacology Stuttgart, Germany ; University of Tuebingen Tuebingen, Germany.
authors.forename: Kathrin
authors.initials: K
authors.lastname: Klein
authors.affiliation: Dr. Margarete Fischer-Bosch-Institute of Clinical Pharmacology Stuttgart, Germany ; University of Tuebingen Tuebingen, Germany ; Department of Clinical Pharmacology, University Hospital Tuebingen Tuebingen, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Schwab
authors.affiliation: Dr. Margarete Fischer-Bosch-Institute of Clinical Pharmacology Stuttgart, Germany ; University of Tuebingen Tuebingen, Germany.
authors.forename: Ulrich M
authors.initials: UM
authors.lastname: Zanger
keywords: HepaRG cells; WNT/β-catenin; chromatin immunoprecipitation; diet–drug interaction; drug metabolism; genotype–phenotype correlation; nuclear receptors; transcriptional regulation