1. Direct evidence for pitavastatin induced chromatin structure change in the KLF4 gene in endothelial cells
- Author
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Maejima, T., Inoue, T., Kanki, Y., Kohro, T., Li, G., Ohta, Y., Kimura, Hiroshi, Kobayashi, M., Taguchi, A., Tsutsumi, S., Iwanari, H., Yamamoto, S., Aruga, H., Dong, S., Stevens, J. F., Poh, H. M., Yamamoto, K., Kawamura, T., Mimura, I., Suehiro, J., Sugiyama, A., Kaneki, K., Shibata, H., Yoshinaka, Y., Doi, T., Asanuma, A., Tanabe, S., Tanaka, T., Minami, T., Hamakubo, T., Sakai, J., Nozaki, N., Aburatani, H., Nangaku, M., Ruan, X., Tanabe, H., Ruan, Y., Ihara, S., Endo, A., Kodama, T., and Wada, Y.
- Subjects
Microarrays ,Thrombomodulin ,lcsh:Medicine ,Gene Expression ,Nitric Oxide Synthase Type III/biosynthesis/genetics ,Vascular Medicine ,Epithelium ,Chromosome conformation capture ,Molecular Cell Biology ,Medicine and Health Sciences ,lcsh:Science ,Regulation of gene expression ,Multidisciplinary ,Chromatin Assembly and Disassembly/*drug effects ,Chromosome Biology ,MEF2 Transcription Factors ,Hydroxymethylglutaryl-CoA Reductase Inhibitors/*pharmacology ,Quinolines/*pharmacology ,Genomics ,Animal Models ,MEF2 Transcription Factors/genetics/metabolism ,Chromatin ,Bioassays and Physiological Analysis ,Thrombomodulin ＜NLM MeSH Terms＞ ,Gene Knockdown Techniques ,Quinolines ,Anatomy ,Cellular Types ,Transcriptome Analysis ,Research Article ,Mef2 ,Chromosome Structure and Function ,Drug Research and Development ,Nitric Oxide Synthase Type III ,Thrombomodulin/biosynthesis/genetics ,Cardiology ,Kruppel-Like Transcription Factors ,Mouse Models ,Kruppel-Like Transcription Factors/*biosynthesis/genetics ,Biology ,Research and Analysis Methods ,Response Elements ,Chromosomes ,Kruppel-Like Factor 4 ,Model Organisms ,Genetics ,Human Umbilical Vein Endothelial Cells ,Humans ,Enhancer ,Transcription factor ,ChIA-PET ,Pharmacology ,lcsh:R ,Biology and Life Sciences ,Endothelial Cells ,Computational Biology ,Epithelial Cells ,Cell Biology ,Genome Analysis ,Atherosclerosis ,Chromatin Assembly and Disassembly ,Molecular biology ,Biological Tissue ,Gene Expression Regulation ,Cardiovascular Anatomy ,Gene Expression Regulation/*drug effects/genetics ,Human Umbilical Vein Endothelial Cells/cytology/*metabolism ,lcsh:Q ,Chromatin/genetics/*metabolism ,Clinical Medicine ,Hydroxymethylglutaryl-CoA Reductase Inhibitors ,Genome Expression Analysis ,Chromatin immunoprecipitation - 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.
- Published
- 2014