title: Krüppel-like factor 9 promotes hepatic cytochrome P450 2D6 expression during pregnancy in CYP2D6-humanized mice.
abstract: Cytochrome P450 2D6 (CYP2D6), a major drug-metabolizing enzyme, is responsible for metabolism of approximately 25% of marketed drugs. Clinical evidence indicates that metabolism of CYP2D6 substrates is increased during pregnancy, but the underlying mechanisms remain unclear. To identify transcription factors potentially responsible for CYP2D6 induction during pregnancy, a panel of genes differentially expressed in the livers of pregnant versus nonpregnant CYP2D6-humanized (tg-CYP2D6) mice was compiled via microarray experiments followed by real-time quantitative reverse-transcription polymerase chain reaction(qRT-PCR) verification. As a result, seven transcription factors-activating transcription factor 5 (ATF5), early growth response 1 (EGR1), forkhead box protein A3 (FOXA3), JUNB, Krüppel-like factor 9 (KLF9), KLF10, and REV-ERBα-were found to be up-regulated in liver during pregnancy. Results from transient transfection and promoter reporter gene assays indicate that KLF9 itself is a weak transactivator of CYP2D6 promoter but significantly enhances CYP2D6 promoter transactivation by hepatocyte nuclear factor 4 (HNF4α), a known transcriptional activator of CYP2D6 expression. The results from deletion and mutation analysis of CYP2D6 promoter activity identified a KLF9 putative binding motif at -22/-14 region to be critical in the potentiation of HNF4α-induced transactivation of CYP2D6. Electrophoretic mobility shift assays revealed a direct binding of KLF9 to the putative KLF binding motif. Results from chromatin immunoprecipitation assay showed increased recruitment of KLF9 to CYP2D6 promoter in the livers of tg-CYP2D6 mice during pregnancy. Taken together, our data suggest that increased KLF9 expression is in part responsible for CYP2D6 induction during pregnancy via the potentiation of HNF4α transactivation of CYP2D6.
authors.affiliation: Department of Pharmacy Practice (K.H.K., H.J.), Department of Biopharmaceutical Sciences (X.P., H.J.), College of Pharmacy, and Department of Pediatrics (W.Z.) and Department of Microbiology and Immunology (A.M.), College of Medicine, University of Illinois at Chicago, Chicago, Illinois; and Laboratory of Epigenetics and Chromatin Dynamics, and Departments of Biology and Molecular Biology, Epigenomics Translational Program, Mayo Clinic Center for Individualized Medicine, Mayo Clinic, Rochester, Minnesota (R.U.).
authors.forename: Kwi Hye
authors.initials: KH
authors.lastname: Koh
authors.affiliation: Department of Pharmacy Practice (K.H.K., H.J.), Department of Biopharmaceutical Sciences (X.P., H.J.), College of Pharmacy, and Department of Pediatrics (W.Z.) and Department of Microbiology and Immunology (A.M.), College of Medicine, University of Illinois at Chicago, Chicago, Illinois; and Laboratory of Epigenetics and Chromatin Dynamics, and Departments of Biology and Molecular Biology, Epigenomics Translational Program, Mayo Clinic Center for Individualized Medicine, Mayo Clinic, Rochester, Minnesota (R.U.).
authors.forename: Xian
authors.initials: X
authors.lastname: Pan
authors.affiliation: Department of Pharmacy Practice (K.H.K., H.J.), Department of Biopharmaceutical Sciences (X.P., H.J.), College of Pharmacy, and Department of Pediatrics (W.Z.) and Department of Microbiology and Immunology (A.M.), College of Medicine, University of Illinois at Chicago, Chicago, Illinois; and Laboratory of Epigenetics and Chromatin Dynamics, and Departments of Biology and Molecular Biology, Epigenomics Translational Program, Mayo Clinic Center for Individualized Medicine, Mayo Clinic, Rochester, Minnesota (R.U.).
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Department of Pharmacy Practice (K.H.K., H.J.), Department of Biopharmaceutical Sciences (X.P., H.J.), College of Pharmacy, and Department of Pediatrics (W.Z.) and Department of Microbiology and Immunology (A.M.), College of Medicine, University of Illinois at Chicago, Chicago, Illinois; and Laboratory of Epigenetics and Chromatin Dynamics, and Departments of Biology and Molecular Biology, Epigenomics Translational Program, Mayo Clinic Center for Individualized Medicine, Mayo Clinic, Rochester, Minnesota (R.U.).
authors.forename: Alan
authors.initials: A
authors.lastname: McLachlan
authors.affiliation: Department of Pharmacy Practice (K.H.K., H.J.), Department of Biopharmaceutical Sciences (X.P., H.J.), College of Pharmacy, and Department of Pediatrics (W.Z.) and Department of Microbiology and Immunology (A.M.), College of Medicine, University of Illinois at Chicago, Chicago, Illinois; and Laboratory of Epigenetics and Chromatin Dynamics, and Departments of Biology and Molecular Biology, Epigenomics Translational Program, Mayo Clinic Center for Individualized Medicine, Mayo Clinic, Rochester, Minnesota (R.U.).
authors.forename: Raul
authors.initials: R
authors.lastname: Urrutia
authors.affiliation: Department of Pharmacy Practice (K.H.K., H.J.), Department of Biopharmaceutical Sciences (X.P., H.J.), College of Pharmacy, and Department of Pediatrics (W.Z.) and Department of Microbiology and Immunology (A.M.), College of Medicine, University of Illinois at Chicago, Chicago, Illinois; and Laboratory of Epigenetics and Chromatin Dynamics, and Departments of Biology and Molecular Biology, Epigenomics Translational Program, Mayo Clinic Center for Individualized Medicine, Mayo Clinic, Rochester, Minnesota (R.U.) yjeong@uic.edu.
authors.forename: Hyunyoung
authors.initials: H
authors.lastname: Jeong
mesh_terms: D000818:Animals; D019389:Cytochrome P-450 CYP2D6 / Q000235:genetics*; D005260:Female; D057809:HEK293 Cells; D051557:Hepatocyte Nuclear Factor 4 / Q000737:chemistry / Q000502:physiology; D006801:Humans; D051741:Kruppel-Like Transcription Factors / Q000737:chemistry / Q000502:physiology*; D008099:Liver / Q000201:enzymology*; D008297:Male; D051379:Mice; D011247:Pregnancy; D011270:Pregnancy, Animal / Q000378:metabolism*; D011401:Promoter Regions, Genetic
chemical_list: D051557:Hepatocyte Nuclear Factor 4; C496593:Hnf4a protein, mouse; C489676:KLF9 protein, human; C489675:Klf9 protein, mouse; D051741:Kruppel-Like Transcription Factors; D019389:Cytochrome P-450 CYP2D6