title: Genome-wide approaches reveal functional interleukin-4-inducible STAT6 binding to the vascular cell adhesion molecule 1 promoter.
abstract: Endothelial cell activation and dysfunction underlie many vascular disorders, including atherosclerosis and inflammation. Here, we show that interleukin-4 (IL-4) markedly induced vascular cell adhesion molecule 1 (VCAM-1), both in cultured endothelial cells and in the intact endothelium in mice. Combined treatment with IL-4 and tumor necrosis factor alpha (TNF-α) resulted in further, sustained induction of VCAM-1 expression. IL-4-mediated induction of VCAM-1 and secondary monocyte adhesion was predominantly regulated by the transcription factor STAT6. Genome-wide survey of IL-4-mediated STAT6 binding from sequential chromatin-immunoprecipitation with deep sequencing (chromatin immunoprecipitation sequencing [ChIP-seq]) in endothelial cells revealed regions of transient and sustained transcription factor binding. Through the combination of DNA microarrays and ChIP-seq at the same time points, the majority of IL-4-responsive genes were shown to be STAT6 dependent and associated with direct STAT6 binding to their promoter. IL-4-mediated stable binding of STAT6 led to sustained target gene expression. Moreover, our strategy led to the identification of a novel functionally important STAT6 binding site within 16 kb upstream of the VCAM-1 gene. Taken together, these findings support a critical role for STAT6 in mediating IL-4 signal transduction in endothelial cells. Identification of a novel IL-4-mediated VCAM-1 enhancer may provide a foundation for targeted therapy in vascular disease.
authors.affiliation: Research Center for Advanced Science and Technology, University of Tokyo, 4-6-1 Komaba, Meguro, Tokyo 153-8904, Japan.
authors.forename: Hideto
authors.initials: H
authors.lastname: Tozawa
authors.forename: Yasuharu
authors.initials: Y
authors.lastname: Kanki
authors.forename: Jun-ichi
authors.initials: J
authors.lastname: Suehiro
authors.forename: Shuichi
authors.initials: S
authors.lastname: Tsutsumi
authors.forename: Takahide
authors.initials: T
authors.lastname: Kohro
authors.forename: Youichiro
authors.initials: Y
authors.lastname: Wada
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Aburatani
authors.forename: William C
authors.initials: WC
authors.lastname: Aird
authors.forename: Tatsuhiko
authors.initials: T
authors.lastname: Kodama
authors.forename: Takashi
authors.initials: T
authors.lastname: Minami
mesh_terms: D000818:Animals; D001665:Binding Sites; D002448:Cell Adhesion; D047369:Chromatin Immunoprecipitation; D042783:Endothelial Cells / Q000378:metabolism; D004727:Endothelium / Q000378:metabolism; D015870:Gene Expression; D057809:HEK293 Cells; D006801:Humans; D015847:Interleukin-4 / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D009000:Monocytes / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012045:Regulatory Sequences, Nucleic Acid; D050800:STAT6 Transcription Factor / Q000378:metabolism*; D015398:Signal Transduction; D014158:Transcription, Genetic; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D019010:Vascular Cell Adhesion Molecule-1 / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism
chemical_list: D050800:STAT6 Transcription Factor; D014409:Tumor Necrosis Factor-alpha; D019010:Vascular Cell Adhesion Molecule-1; D015847:Interleukin-4