Rifampin modulation of xeno- and endobiotic conjugating enzyme mRNA expression and associated microRNAs in human hepatocytes

Brandon T. Gufford, Jason D. Robarge, Michael Eadon, Hongyu Gao, Hai Lin, Yunlong Liu, Zeruesenay Desta, Todd Skaar

Research output: Contribution to journalArticle

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Abstract

Rifampin is a pleiotropic inducer of multiple drug metabolizing enzymes and transporters. This work utilized a global approach to evaluate rifampin effects on conjugating enzyme gene expression with relevance to human xeno- and endo-biotic metabolism. Primary human hepatocytes from 7 subjects were treated with rifampin (10 μmol/L, 24 hours). Standard methods for RNA-seq library construction, EZBead preparation, and NextGen sequencing were used to measure UDP-glucuronosyl transferase UGT, sulfonyltransferase SULT, N acetyltransferase NAT, and glutathione-S-transferase GST mRNA expression compared to vehicle control (0.01% MeOH). Rifampin-induced (>1.25-fold) mRNA expression of 13 clinically important phase II drug metabolizing genes and repressed (>1.25-fold) the expression of 3 genes (P <.05). Rifampin-induced miRNA expression changes correlated with mRNA changes and miRNAs were identified that may modulate conjugating enzyme expression. NAT2 gene expression was most strongly repressed (1.3-fold) by rifampin while UGT1A4 and UGT1A1 genes were most strongly induced (7.9- and 4.8-fold, respectively). Physiologically based pharmacokinetic modeling (PBPK) was used to simulate the clinical consequences of rifampin induction of CYP3A4- and UGT1A4-mediated midazolam metabolism. Simulations evaluating isolated UGT1A4 induction predicted increased midazolam N-glucuronide exposure (~4-fold) with minimal reductions in parent midazolam exposure (~10%). Simulations accounting for simultaneous induction of both CYP3A4 and UGT1A4 predicted a ~10-fold decrease in parent midazolam exposure with only a ~2-fold decrease in midazolam N-glucuronide metabolite exposure. These data reveal differential effects of rifampin on the human conjugating enzyme transcriptome and potential associations with miRNAs that form the basis for future mechanistic studies to elucidate the interplay of conjugating enzyme regulatory elements.

Original languageEnglish (US)
Article numbere00386
JournalPharmacology Research and Perspectives
Volume6
Issue number2
DOIs
StatePublished - Apr 1 2018

Fingerprint

Xenobiotics
Rifampin
MicroRNAs
Hepatocytes
Midazolam
Messenger RNA
Enzymes
Cytochrome P-450 CYP3A
Glucuronides
Gene Expression
Acetyltransferases
Uridine Diphosphate
Transferases
Glutathione Transferase
Transcriptome
Pharmaceutical Preparations
Genes
Libraries
Pharmacokinetics
RNA

Keywords

  • drug metabolizing enzyme induction
  • miRNA modulation of mRNA
  • PBPK modeling
  • phase 2 enzyme induction
  • rifampin miRNA induction
  • rifampin mRNA repression

ASJC Scopus subject areas

  • Neurology
  • Pharmacology, Toxicology and Pharmaceutics(all)

Cite this

Rifampin modulation of xeno- and endobiotic conjugating enzyme mRNA expression and associated microRNAs in human hepatocytes. / Gufford, Brandon T.; Robarge, Jason D.; Eadon, Michael; Gao, Hongyu; Lin, Hai; Liu, Yunlong; Desta, Zeruesenay; Skaar, Todd.

In: Pharmacology Research and Perspectives, Vol. 6, No. 2, e00386, 01.04.2018.

Research output: Contribution to journalArticle

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