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: Jessica K Rieger, Sandra Reutter, Ute Hofmann, Matthias Schwab, Ulrich M Zanger
• Journal: Drug metabolism and disposition: the biological fate of chemicals (Drug Metab Dispos) Vol. 43 Issue 6 Pg. 884-8 (Jun 2015) ISSN: 1521-009X [Electronic] United States
• PMID: 25802328 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2015 by The American Society for Pharmacology and Experimental Therapeutics.

Chemical References

• 3' Untranslated Regions
• MIRN130 microRNA, human
• MicroRNAs
• RNA, Messenger
• RNA, Small Interfering
• Recombinant Fusion Proteins
• CYP2C9 protein, human
• Cytochrome P-450 CYP2C9

Topics

• 3' Untranslated Regions
• Cell Line
• Cholestasis, Intrahepatic (enzymology, immunology, metabolism)
• Computational Biology
• Cytochrome P-450 CYP2C9 (chemistry, genetics, metabolism)
• Enzyme Repression
• Genes, Reporter
• Hepatitis (enzymology, immunology, metabolism)
• Hepatocytes (enzymology, immunology, metabolism)
• Humans
• MicroRNAs (antagonists & inhibitors, chemistry, metabolism)
• Models, Biological
• RNA Interference
• RNA, Messenger (metabolism)
• RNA, Small Interfering
• Recombinant Fusion Proteins (chemistry, metabolism)
• Up-Regulation