title: Direct evidence for pitavastatin induced chromatin structure change in the KLF4 gene in endothelial cells.
abstract: Statins exert atheroprotective effects through the induction of specific transcriptional factors in multiple organs. In endothelial cells, statin-dependent atheroprotective gene up-regulation is mediated by Kruppel-like factor (KLF) family transcription factors. To dissect the mechanism of gene regulation, we sought to determine molecular targets by performing microarray analyses of human umbilical vein endothelial cells (HUVECs) treated with pitavastatin, and KLF4 was determined to be the most highly induced gene. In addition, it was revealed that the atheroprotective genes induced with pitavastatin, such as nitric oxide synthase 3 (NOS3) and thrombomodulin (THBD), were suppressed by KLF4 knockdown. Myocyte enhancer factor-2 (MEF2) family activation is reported to be involved in pitavastatin-dependent KLF4 induction. We focused on MEF2C among the MEF2 family members and identified a novel functional MEF2C binding site 148 kb upstream of the KLF4 gene by chromatin immunoprecipitation along with deep sequencing (ChIP-seq) followed by luciferase assay. By applying whole genome and quantitative chromatin conformation analysis {chromatin interaction analysis with paired end tag sequencing (ChIA-PET), and real time chromosome conformation capture (3C) assay}, we observed that the MEF2C-bound enhancer and transcription start site (TSS) of KLF4 came into closer spatial proximity by pitavastatin treatment. 3D-Fluorescence in situ hybridization (FISH) imaging supported the conformational change in individual cells. Taken together, dynamic chromatin conformation change was shown to mediate pitavastatin-responsive gene induction in endothelial cells.
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, Meguro-ku, Tokyo, Japan; Tokyo New Drug Research Laboratories, Kowa Company Ltd., Higashimurayama, Tokyo, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Maejima
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, Meguro-ku, Tokyo, Japan; Division of Nephrology and Endocrinology, The University of Tokyo School of Medicine, Bunkyo-ku, Tokyo, Japan.
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Inoue
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, Meguro-ku, Tokyo, Japan.
authors.forename: Yasuharu
authors.initials: Y
authors.lastname: Kanki
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, Meguro-ku, Tokyo, Japan; Department of Translational Research for Healthcare and Clinical Science, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo, Japan.
authors.forename: Takahide
authors.initials: T
authors.lastname: Kohro
authors.affiliation: Jackson Laboratory for Genomic Medicine, Farmington, Connecticut, United States of America.
authors.forename: Guoliang
authors.initials: G
authors.lastname: Li
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, Meguro-ku, Tokyo, Japan.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Ohta
authors.affiliation: Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kimura
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, Meguro-ku, Tokyo, Japan.
authors.forename: Mika
authors.initials: M
authors.lastname: Kobayashi
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, Meguro-ku, Tokyo, Japan.
authors.forename: Akashi
authors.initials: A
authors.lastname: Taguchi
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, Meguro-ku, Tokyo, Japan.
authors.forename: Shuichi
authors.initials: S
authors.lastname: Tsutsumi
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, Meguro-ku, Tokyo, Japan.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Iwanari
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, Meguro-ku, Tokyo, Japan.
authors.forename: Shogo
authors.initials: S
authors.lastname: Yamamoto
authors.affiliation: Thermo Fisher Scientific, South San Francisco, California, United States of America.
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Aruga
authors.affiliation: Thermo Fisher Scientific, South San Francisco, California, United States of America.
authors.forename: Shoulian
authors.initials: S
authors.lastname: Dong
authors.affiliation: Thermo Fisher Scientific, South San Francisco, California, United States of America.
authors.forename: Junko F
authors.initials: JF
authors.lastname: Stevens
authors.affiliation: Jackson Laboratory for Genomic Medicine, Farmington, Connecticut, United States of America.
authors.forename: Huay Mei
authors.initials: HM
authors.lastname: Poh
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, Meguro-ku, Tokyo, Japan.
authors.forename: Kazuki
authors.initials: K
authors.lastname: Yamamoto
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, Meguro-ku, Tokyo, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Kawamura
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, Meguro-ku, Tokyo, Japan; Division of Nephrology and Endocrinology, The University of Tokyo School of Medicine, Bunkyo-ku, Tokyo, Japan.
authors.forename: Imari
authors.initials: I
authors.lastname: Mimura
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, Meguro-ku, Tokyo, Japan.
authors.forename: Jun-ichi
authors.initials: J
authors.lastname: Suehiro
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, Meguro-ku, Tokyo, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Sugiyama
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, Meguro-ku, Tokyo, Japan.
authors.forename: Kiyomi
authors.initials: K
authors.lastname: Kaneki
authors.affiliation: Tokyo New Drug Research Laboratories, Kowa Company Ltd., Higashimurayama, Tokyo, Japan.
authors.forename: Haruki
authors.initials: H
authors.lastname: Shibata
authors.affiliation: Tokyo New Drug Research Laboratories, Kowa Company Ltd., Higashimurayama, Tokyo, Japan.
authors.forename: Yasunobu
authors.initials: Y
authors.lastname: Yoshinaka
authors.affiliation: Tokyo New Drug Research Laboratories, Kowa Company Ltd., Higashimurayama, Tokyo, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Doi
authors.affiliation: Tokyo New Drug Research Laboratories, Kowa Company Ltd., Higashimurayama, Tokyo, Japan.
authors.forename: Akimune
authors.initials: A
authors.lastname: Asanuma
authors.affiliation: Tokyo New Drug Research Laboratories, Kowa Company Ltd., Higashimurayama, Tokyo, Japan.
authors.forename: Sohei
authors.initials: S
authors.lastname: Tanabe
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, Meguro-ku, Tokyo, Japan.
authors.forename: Toshiya
authors.initials: T
authors.lastname: Tanaka
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, Meguro-ku, Tokyo, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Minami
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, Meguro-ku, Tokyo, Japan.
authors.forename: Takao
authors.initials: T
authors.lastname: Hamakubo
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, Meguro-ku, Tokyo, Japan.
authors.forename: Juro
authors.initials: J
authors.lastname: Sakai
authors.affiliation: Mab Institute Inc., North Advancement Center for Science & Technology, Sapporo, Hokkaido, Japan.
authors.forename: Naohito
authors.initials: N
authors.lastname: Nozaki
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, Meguro-ku, Tokyo, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Aburatani
authors.affiliation: Division of Nephrology and Endocrinology, The University of Tokyo School of Medicine, Bunkyo-ku, Tokyo, Japan.
authors.forename: Masaomi
authors.initials: M
authors.lastname: Nangaku
authors.affiliation: Genome Institute of Singapore, Singapore, Singapore.
authors.forename: Xiaoan
authors.initials: X
authors.lastname: Ruan
authors.affiliation: Department of Evolutionary Studies of Biosystems, School of Advanced Sciences, The Graduate University for Advanced Studies (Sokendai), Hayama, Kanagawa, Japan.
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Tanabe
authors.affiliation: Jackson Laboratory for Genomic Medicine, Farmington, Connecticut, United States of America.
authors.forename: Yijun
authors.initials: Y
authors.lastname: Ruan
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, Meguro-ku, Tokyo, Japan.
authors.forename: Sigeo
authors.initials: S
authors.lastname: Ihara
authors.affiliation: Biopharm Research Laboratories, Inc., Koganei, Tokyo, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Endo
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, Meguro-ku, Tokyo, Japan.
authors.forename: Tatsuhiko
authors.initials: T
authors.lastname: Kodama
authors.affiliation: Research Center for Advanced Science and Technology, The University of Tokyo, Meguro-ku, Tokyo, Japan; Radioisotope Center, The University of Tokyo, Bunkyo-ku, Tokyo, Japan.
authors.forename: Youichiro
authors.initials: Y
authors.lastname: Wada
mesh_terms: D002843:Chromatin / Q000235:genetics / Q000378:metabolism*; D042002:Chromatin Assembly and Disassembly / Q000187:drug effects*; D005786:Gene Expression Regulation / Q000187:drug effects* / Q000235:genetics; D055785:Gene Knockdown Techniques; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors / Q000494:pharmacology*; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000096:biosynthesis* / Q000235:genetics; D064326:MEF2 Transcription Factors / Q000235:genetics / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000096:biosynthesis / Q000235:genetics; D011804:Quinolines / Q000494:pharmacology*; D020218:Response Elements; D018180:Thrombomodulin / Q000096:biosynthesis / Q000235:genetics
chemical_list: D002843:Chromatin; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; C000715467:KLF4 protein, human; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; D064326:MEF2 Transcription Factors; C578040:MEF2C protein, human; D011804:Quinolines; D018180:Thrombomodulin; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III; C108475:pitavastatin