title: Genome-wide approaches reveal functional vascular endothelial growth factor (VEGF)-inducible nuclear factor of activated T cells (NFAT) c1 binding to angiogenesis-related genes in the endothelium.
abstract: VEGF is a key regulator of endothelial cell migration, proliferation, and inflammation, which leads to activation of several signaling cascades, including the calcineurin-nuclear factor of activated T cells (NFAT) pathway. NFAT is not only important for immune responses but also for cardiovascular development and the pathogenesis of Down syndrome. By using Down syndrome model mice and clinical patient samples, we showed recently that the VEGF-calcineurin-NFAT signaling axis regulates tumor angiogenesis and tumor metastasis. However, the connection between genome-wide views of NFAT-mediated gene regulation and downstream gene function in the endothelium has not been studied extensively. Here we performed comprehensive mapping of genome-wide NFATc1 binding in VEGF-stimulated primary cultured endothelial cells and elucidated the functional consequences of VEGF-NFATc1-mediated phenotypic changes. A comparison of the NFATc1 ChIP sequence profile and epigenetic histone marks revealed that predominant NFATc1-occupied peaks overlapped with promoter-associated histone marks. Moreover, we identified two novel NFATc1 regulated genes, CXCR7 and RND1. CXCR7 knockdown abrogated SDF-1- and VEGF-mediated cell migration and tube formation. siRNA treatment of RND1 impaired vascular barrier function, caused RhoA hyperactivation, and further stimulated VEGF-mediated vascular outgrowth from aortic rings. Taken together, these findings suggest that dynamic NFATc1 binding to target genes is critical for VEGF-mediated endothelial cell activation. CXCR7 and RND1 are NFATc1 target genes with multiple functions, including regulation of cell migration, tube formation, and barrier formation in endothelial cells.
authors.affiliation: From the Division of Vascular Biology.
authors.forename: Jun-ichi
authors.initials: J
authors.lastname: Suehiro
authors.affiliation: From the Division of Vascular Biology, Systems Biology, The Research Center for Advanced Science and Technology, The University of Tokyo, Tokyo, 153-8904 Japan and.
authors.forename: Yasuharu
authors.initials: Y
authors.lastname: Kanki
authors.affiliation: From the Division of Vascular Biology.
authors.forename: Chihiro
authors.initials: C
authors.lastname: Makihara
authors.affiliation: From the Division of Vascular Biology, the Department of Cancer Biology, Abramson Family Cancer Research Institute, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104.
authors.forename: Keri
authors.initials: K
authors.lastname: Schadler
authors.affiliation: From the Division of Vascular Biology.
authors.forename: Mai
authors.initials: M
authors.lastname: Miura
authors.affiliation: From the Division of Vascular Biology.
authors.forename: Yuuka
authors.initials: Y
authors.lastname: Manabe
authors.affiliation: Genome Science Division, and.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Aburatani
authors.affiliation: Systems Biology, The Research Center for Advanced Science and Technology, The University of Tokyo, Tokyo, 153-8904 Japan and.
authors.forename: Tatsuhiko
authors.initials: T
authors.lastname: Kodama
authors.affiliation: From the Division of Vascular Biology, minami@med.rcast.u-tokyo.ac.jp.
authors.forename: Takashi
authors.initials: T
authors.lastname: Minami
mesh_terms: D000818:Animals; D019556:COS Cells; D002465:Cell Movement; D002522:Chlorocebus aethiops; D018920:Coculture Techniques; D042783:Endothelial Cells / Q000166:cytology; D004730:Endothelium, Vascular / Q000378:metabolism*; D044127:Epigenesis, Genetic; D005347:Fibroblasts / Q000378:metabolism; D055106:Genome-Wide Association Study; D057809:HEK293 Cells; D006706:Homeostasis; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D051379:Mice; D050778:NFATC Transcription Factors / Q000378:metabolism*; D009389:Neovascularization, Pathologic; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D019718:Receptors, CXCR4 / Q000378:metabolism; D015398:Signal Transduction; D015533:Transcriptional Activation; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*; D020741:rho GTP-Binding Proteins / Q000378:metabolism
chemical_list: C527276:CXCR4 protein, human; D050778:NFATC Transcription Factors; C494003:NFATC1 protein, human; C491772:RND1 protein, human; D019718:Receptors, CXCR4; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D020741:rho GTP-Binding Proteins
keywords: Angiogenesis; ChIP-seq; Endothelial Cell; Gene Transcription; Genomics; Microarray; NFAT Transcription Factor; Signaling; Vascular Biology; Vascular Endothelial Growth Factor (VEGF)