title: Inflammation-associated microRNA-130b down-regulates cytochrome P450 activities and directly targets CYP2C9.
abstract: Expression of genes involved in absorption, distribution, metabolism, and excretion (ADME) of drugs is impaired in pathophysiologic conditions such as cholestasis and inflammation. The mechanisms of ADME gene down-regulation remain unclear. In our previous study, strongly elevated levels of microRNAs (miRNA) miR-21, miR-34a, and miR-130b in cholestatic liver and of miR-21 and miR-130b during inflammation were observed. Using HepaRG cells, which retain many functional characteristics of human hepatocytes, we investigated the potential of these miRNAs to down-regulate ADME genes. Cells were transfected with the corresponding miRNA mimics, chemically modified double-stranded RNAs that mimic endogenous miRNAs, followed by mRNA profiling by quantitative reverse-transcription polymerase chain reaction. Activities of six cytochrome P450 enzymes (CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, and CYP3A4) were determined with a liquid chromatography with tandem mass spectrometric cocktail assay. Although miR-21 and miR-34a showed few effects, transfection of miR-130b led to significantly lower expression of nuclear receptors constitutive androstane receptor (CAR) and farnesoid X receptor (FXRα), the CYPs 1A1, 1A2, 2A6, 2C8, 2C9, and 2C19, as well as GSTA2. Furthermore, miR-130b negatively affected activity levels of all measured P450s by at least 30%. Reporter gene assays employing the CYP2C9 3'-untranslated region (3'-UTR) confirmed direct regulation by miR-130b. These data support miR-130b as a potential negative regulator of drug metabolism by directly and/or indirectly affecting the expression of several ADME genes. This may be of relevance in pathophysiologic conditions such as cholestasis and inflammation, which are associated with increased miR-130b expression.
authors: Dr. Margarete Fischer-Bosch Institute of Clinical Pharmacology (J.K.R., S.R., U.H., M.S., U.M.Z.) and Department of Clinical Pharmacology (M.S.), University of Tuebingen, Tuebingen, Germany.
authors: Jessica K
authors: JK
authors: Rieger
authors: Dr. Margarete Fischer-Bosch Institute of Clinical Pharmacology (J.K.R., S.R., U.H., M.S., U.M.Z.) and Department of Clinical Pharmacology (M.S.), University of Tuebingen, Tuebingen, Germany.
authors: Sandra
authors: S
authors: Reutter
authors: Dr. Margarete Fischer-Bosch Institute of Clinical Pharmacology (J.K.R., S.R., U.H., M.S., U.M.Z.) and Department of Clinical Pharmacology (M.S.), University of Tuebingen, Tuebingen, Germany.
authors: Ute
authors: U
authors: Hofmann
authors: Dr. Margarete Fischer-Bosch Institute of Clinical Pharmacology (J.K.R., S.R., U.H., M.S., U.M.Z.) and Department of Clinical Pharmacology (M.S.), University of Tuebingen, Tuebingen, Germany.
authors: Matthias
authors: M
authors: Schwab
authors: Dr. Margarete Fischer-Bosch Institute of Clinical Pharmacology (J.K.R., S.R., U.H., M.S., U.M.Z.) and Department of Clinical Pharmacology (M.S.), University of Tuebingen, Tuebingen, Germany uli.zanger@ikp-stuttgart.de.
authors: Ulrich M
authors: UM
authors: Zanger
mesh_terms: D020413:3' Untranslated Regions; D002460:Cell Line; D002780:Cholestasis, Intrahepatic / Q000201:enzymology / Q000276:immunology / Q000378:metabolism; D019295:Computational Biology; D065729:Cytochrome P-450 CYP2C9 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004794:Enzyme Repression; D017930:Genes, Reporter; D006505:Hepatitis / Q000201:enzymology / Q000276:immunology / Q000378:metabolism; D022781:Hepatocytes / Q000201:enzymology / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D035683:MicroRNAs / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D008954:Models, Biological; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D015854:Up-Regulation
chemical_list: D020413:3' Untranslated Regions; C554019:MIRN130 microRNA, human; D035683:MicroRNAs; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; C450260:CYP2C9 protein, human; D065729:Cytochrome P-450 CYP2C9