Open AccessRifampin modulation of xeno‐ and endobiotic conjugating enzyme mRNA expression and associated micro RNA s in human hepatocytes
Author(s) -
Gufford Brandon T.,
Robarge Jason D.,
Eadon Michael T.,
Gao Hongyu,
Lin Hai,
Liu Yunlong,
Desta Zeruesenay,
Skaar Todd C.
Publication year - 2018
Publication title -
pharmacology research and perspectives
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.975
H-Index - 27
ISSN - 2052-1707
DOI - 10.1002/prp2.386
Subject(s) - cyp3a4 , gene expression , enzyme , biology , messenger rna , gene , constitutive androstane receptor , drug metabolism , microbiology and biotechnology , chemistry , pharmacology , biochemistry , cytochrome p450 , transcription factor , nuclear receptor
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, EZB ead 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% Me OH ). 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 mi RNA expression changes correlated with mRNA changes and mi RNA s were identified that may modulate conjugating enzyme expression. NAT 2 gene expression was most strongly repressed (1.3‐fold) by rifampin while UGT 1A4 and UGT 1A1 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 CYP 3A4‐ and UGT 1A4‐mediated midazolam metabolism. Simulations evaluating isolated UGT 1A4 induction predicted increased midazolam N‐glucuronide exposure (~4‐fold) with minimal reductions in parent midazolam exposure (~10%). Simulations accounting for simultaneous induction of both CYP 3A4 and UGT 1A4 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 mi RNA s that form the basis for future mechanistic studies to elucidate the interplay of conjugating enzyme regulatory elements.