title: Bone morphogenetic protein 4 regulates microRNAs miR-494 and miR-126-5p in control of endothelial cell function in angiogenesis.
abstract: MicroRNAs are small non-coding RNAs that negatively regulate posttranscriptional gene expression. Several microRNAs have been described to regulate the process of angiogenesis. Previously, we have shown that bone morphogenetic protein 4 (BMP4) increased the pro-angiogenic activity of endothelial cells. In this project, we now investigated how the pro-angiogenic BMP4 effect is mediated by microRNAs. First, we performed a microRNA array with BMP4-stimulated human umbilical vein endothelial cells (HUVECs). Among the top-regulated microRNAs, we detected a decreased expression of miR-494 and increased expression of miR-126-5p. Next, we analysed the canonical Smad and alternative signalling pathways, through which BMP4 would regulate miR-126-5p and miR-494 expression. Furthermore, the functional effect of miR-494 and miR-126-5p on endothelial cells was investigated. MicroRNA-494 overexpression decreased endothelial cell proliferation, migration and sprout formation. Consistently, miR-494 inhibition increased endothelial cell function. As potential miR-494 targets, bFGF and BMP endothelial cell precursor-derived regulator (BMPER) were identified and confirmed by western blot. Luciferase assays showed direct miR-494 binding in BMPER 3'UTR. In contrast, miR-126-5p overexpression increased pro-angiogenic endothelial cell behaviour and, accordingly, miR-126-5p inhibition decreased endothelial cell function. As a direct miR-126-5p target we identified the anti-angiogenic thrombospondin-1 which was confirmed by western blot analysis and luciferase assays. In the Matrigel plug assay application of antagomiR-494 increased endothelial cell ingrowth, whereas antagomiR-126-5p treatment decreased cell ingrowth in vivo. Taken together, through differential regulation of the anti-angiomiR-494 and the angiomiR-126-5p by BMP4 both microRNAs contribute to the pro-angiogenic BMP4 effect on endothelial cells.
authors.affiliation: Jennifer Susanne Esser, PhD, University Heart Center Freiburg, Department of Cardiology and Angiology I, Cardiovascular Biology Group, Breisacher Str. 33, 79106 Freiburg, Germany, Tel.: +49 76127070440, Fax: +49 76127070450, E-mail: Jennifer.Esser@universitaets-herzzentrum.de.
authors.forename: Jennifer S
authors.initials: JS
authors.lastname: Esser
authors.forename: Erika
authors.initials: E
authors.lastname: Saretzki
authors.forename: Franziska
authors.initials: F
authors.lastname: Pankratz
authors.forename: Bianca
authors.initials: B
authors.lastname: Engert
authors.forename: Sebastian
authors.initials: S
authors.lastname: Grundmann
authors.forename: Christoph
authors.initials: C
authors.lastname: Bode
authors.forename: Martin
authors.initials: M
authors.lastname: Moser
authors.forename: Qian
authors.initials: Q
authors.lastname: Zhou
mesh_terms: D020413:3' Untranslated Regions; D043925:Angiogenesis Inducing Agents / Q000494:pharmacology*; D000818:Animals; D001665:Binding Sites; D055415:Bone Morphogenetic Protein 4 / Q000494:pharmacology*; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D016222:Fibroblast Growth Factor 2 / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D051379:Mice; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D041681:NIH 3T3 Cells; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D015398:Signal Transduction / Q000187:drug effects; D019700:Thrombospondin 1 / Q000235:genetics / Q000378:metabolism; D014162:Transfection
chemical_list: D020413:3' Untranslated Regions; D043925:Angiogenesis Inducing Agents; C525809:BMP4 protein, human; C476724:BMPER protein, human; D055415:Bone Morphogenetic Protein 4; D002352:Carrier Proteins; C527111:MIRN126 microRNA, human; C547168:MIRN494 microRNA, human; D035683:MicroRNAs; D019700:Thrombospondin 1; C000611726:thrombospondin-1, human; D016222:Fibroblast Growth Factor 2
keywords: BMP4; angiogenesis; endothelial cells; microRNAs