Identification of a potent endothelium-derived angiogenic factor

Abstract

Los nombres de dos de los autores son incorrectos. Los nombres del noveno y décimo autor son respectivamente: Alberto Ortíz y María Dolores Sánchez-Niño. La abreviatura correcta en contribuciones del décimo autor es MDSN. La corrección se publicó en: PLoS ONE 8(8): 10.1371/annotation/0c4c8143-c10e-4fe0-b944-b1fcd79995e7. ; The secretion of angiogenic factors by vascular endothelial cells is one of the key mechanisms of angiogenesis. Here we report on the isolation of a new potent angiogenic factor, diuridine tetraphosphate (Up4U) from the secretome of human endothelial cells. The angiogenic effect of the endothelial secretome was partially reduced after incubation with alkaline phosphatase and abolished in the presence of suramin. In one fraction, purified to homogeneity by reversed phase and affinity chromatography, Up4U was identified by MALDI-LIFT-fragment-mass-spectrometry, enzymatic cleavage analysis and retention-time comparison. Beside a strong angiogenic effect on the yolk sac membrane and the developing rat embryo itself, Up4U increased the proliferation rate of endothelial cells and, in the presence of PDGF, of vascular smooth muscle cells. Up4U stimulated the migration rate of endothelial cells via P2Y2-receptors, increased the ability of endothelial cells to form capillary-like tubes and acts as a potent inducer of sprouting angiogenesis originating from gel-embedded EC spheroids. Endothelial cells released Up4U after stimulation with shear stress. Mean total plasma Up4U concentrations of healthy subjects (N = 6) were sufficient to induce angiogenic and proliferative effects (1.3460.26 nmol L-1). In conclusion, Up4U is a novel strong human endothelium-derived angiogenic factor. ; This study was supported by a grant from the German Research Foundation (DFG, Ja-972/11-1; JJ), the Sonnenfeld Foundation (JJ; VJ) and Federal Ministry of Education and Research (BMBF; 01GR0807 (JJ)). The author (VJ; JJ) was supported by grants FP7-HEALTH24 2009-2.4.5-2 to "SYSKID" (241544) and "HEALTH 2011.2.4.2-2″ to "Mascara" (278249) from the European Union. Eutox, FIS PS09/00447, ISCIII-RETIC REDinREN/RD12/0021/0001 and 0004. Comunidad de Madrid/S2010/BMD-2378(NMDS; AAO); FIS-Sara Borrell to MDSN, Programa Intensificación Actividad Investigadora (ISCIII/Agencia Laín-Entralgo/CM) (AAO). VJ received a grant from the Else-Kroener-foundation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.