title: Analysis of Glycogen Synthase Kinase Inhibitors That Regulate Cytochrome P450 Expression in Primary Human Hepatocytes by Activation of β-Catenin, Aryl Hydrocarbon Receptor and Pregnane X Receptor Signaling.
abstract: Cytochrome P450 (CYP) expression and activity are not homogeneous in the liver lobules. Indeed, CYPs are mainly expressed and induced in centrilobular hepatocytes. The wingless-type MMTV integration site family (WNT)/β-catenin pathway was identified as a major regulator of this zonal organization. We have recently demonstrated that in primary human hepatocytes (PHHs), the expression of CYP2E1, CYP1A2, and aryl hydrocarbon receptor (AhR), but not of CYP3A4, is regulated by the WNT/β-catenin pathway in response to WNT3a, its canonical activator. Here, we investigated whether glycogen synthase kinase 3β (GSK3β) inhibitors, which mimic the action of WNT molecules, could be used in PHHs to activate the β-catenin pathway to study CYP expression. We assessed the activity of 6BIO (6-bromoindirubin-3'-oxime), CHIR99021 (6-((2-((4-(2,4-dichlorophenyl)-5-(4methyl-1H-imidazol-2-yl)pyrimidin-2-yl)amino)ethyl)amino) nicotinonitrile), and GSK3iXV (Pyridocarbazolo-cyclopentadienyl Ruthenium complex GSK3 inhibitor XV) that belong to structurally different families of GSK3β inhibitors. Using small interfering RNAs, reporter gene assays, and molecular docking predictions, we demonstrated that GSK3β inhibitors can activate the WNT/β-catenin pathway in PHHs to regulate CYP2E1 expression. We also found that 6BIO and GSK3iXV are AhR full agonists that participate, through AhR signaling, to CYP1A2 induction. Conversely, CHIR99021 is an AhR partial agonist, and a pregnane X receptor ligand and partial agonist, thus regulating CYP1A2 and CYP3A4 gene expression in a β-catenin-independent manner. In conclusion, GSK3β inhibitors can activate the WNT/β-catenin pathway in PHHs. Nevertheless, their role in CYP regulation should be analyzed with caution as these molecules can interact with xenosensors.
authors.affiliation: *INSERM, U1183, Institut de Recherche en Médecine régénérative et Biothérapie, Montpellier F-34295, France; Université de Montpellier, UMR 1183, Montpellier F-34295, France;
authors.forename: Philippe
authors.initials: P
authors.lastname: Briolotti
authors.affiliation: CNRS, FRE 3689 - Université de Montpellier, Centre d'études d'agents Pathogènes et Biotechnologies pour la Santé (CPBS), Montpellier F-34293, France;
authors.forename: Laurent
authors.initials: L
authors.lastname: Chaloin
authors.affiliation: Institut de Recherche en Cancérologie de Montpellier, Montpellier F-34298, France;
authors.forename: Patrick
authors.initials: P
authors.lastname: Balaguer
authors.affiliation: Sanofi Aventis, Drug Disposition Domain, Montpellier F-34184, France;
authors.forename: Franck
authors.initials: F
authors.lastname: Da Silva
authors.affiliation: Faculty of Medicine and Dentistry, Department of Medical Chemistry and Biochemistry, Palacky University, Olomouc, Czech Republic;
authors.forename: Veronika
authors.initials: V
authors.lastname: Tománková
authors.affiliation: CNRS 5203 - INSERM 1191, Institut de Génomique Fonctionnelle, Montpellier F-34094, France;
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Pascussi
authors.affiliation: *INSERM, U1183, Institut de Recherche en Médecine régénérative et Biothérapie, Montpellier F-34295, France; Université de Montpellier, UMR 1183, Montpellier F-34295, France;
authors.forename: Cédric
authors.initials: C
authors.lastname: Duret
authors.affiliation: Département de chirurgie digestive, CHU Saint Eloi, Montpellier F-34295, France;
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Fabre
authors.affiliation: Département d'anatomopathologie, CHU Gui de Chauliac, Montpellier F-34295, France; and.
authors.forename: Jeanne
authors.initials: J
authors.lastname: Ramos
authors.affiliation: Sanofi Aventis, Drug Disposition Domain, Montpellier F-34184, France;
authors.forename: Sylvie
authors.initials: S
authors.lastname: Klieber
authors.affiliation: *INSERM, U1183, Institut de Recherche en Médecine régénérative et Biothérapie, Montpellier F-34295, France; Université de Montpellier, UMR 1183, Montpellier F-34295, France;
authors.forename: Patrick
authors.initials: P
authors.lastname: Maurel
authors.affiliation: *INSERM, U1183, Institut de Recherche en Médecine régénérative et Biothérapie, Montpellier F-34295, France; Université de Montpellier, UMR 1183, Montpellier F-34295, France; **CHU Montpellier, Institut de Recherche en Médecine régénérative et Biothérapie, Montpellier F-34295, France.
authors.forename: Martine
authors.initials: M
authors.lastname: Daujat-Chavanieu
authors.affiliation: *INSERM, U1183, Institut de Recherche en Médecine régénérative et Biothérapie, Montpellier F-34295, France; Université de Montpellier, UMR 1183, Montpellier F-34295, France; sabine.gerbal-chaloin@inserm.fr.
authors.forename: Sabine
authors.initials: S
authors.lastname: Gerbal-Chaloin
mesh_terms: D051792:Basic Helix-Loop-Helix Proteins / Q000819:agonists* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D065693:Cytochrome P-450 Enzyme Inducers / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004790:Enzyme Induction / Q000187:drug effects; D005260:Female; D017930:Genes, Reporter / Q000187:drug effects; D038362:Glycogen Synthase Kinase 3 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D000071679:Glycogen Synthase Kinase 3 beta; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D008297:Male; D062105:Molecular Docking Simulation; D009942:Organometallic Compounds / Q000494:pharmacology; D010091:Oximes / Q000494:pharmacology; D000077297:Pregnane X Receptor; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D011725:Pyridines / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D034622:RNA Interference; D018336:Receptors, Aryl Hydrocarbon / Q000819:agonists* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011987:Receptors, Steroid / Q000819:agonists* / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D060449:Wnt Signaling Pathway / Q000187:drug effects; D051176:beta Catenin / Q000819:agonists* / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D065693:Cytochrome P-450 Enzyme Inducers; D003577:Cytochrome P-450 Enzyme System; D038362:Glycogen Synthase Kinase 3; D000071679:Glycogen Synthase Kinase 3 beta; D007211:Indoles; D009942:Organometallic Compounds; D010091:Oximes; D000077297:Pregnane X Receptor; D047428:Protein Kinase Inhibitors; D011725:Pyridines; D011743:Pyrimidines; D018336:Receptors, Aryl Hydrocarbon; D011987:Receptors, Steroid; D011993:Recombinant Fusion Proteins; D051176:beta Catenin; C483321:6-bromoindirubin-3'-oxime; C111235:AHR protein, human; C495270:CTNNB1 protein, human; C473711:Chir 99021; C000609764:GSK3iXV; C000605506:GSK3B protein, human
keywords: AhR; CTNNB1; CYP; GSK3β; PXR; crosstalk