Identification of a Potent Endothelium-Derived Angiogenic Factor

• Published in: PloS one Vol. 8; no. 7; p. e68575
• Main Authors: Jankowski, Vera, Tölle, Markus, Tran, Thi Nguyet Anh, van der Giet, Markus, Schuchardt, Mirjam, Lehmann, Kerstin, Janke, Doreen, Flick, Burkhard, Ortiz, Alberto Arduan, Sanchez, Niño Maria Dolores, Tepel, Martin, Zidek, Walter, Jankowski, Joachim
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 29.07.2013; Public Library of Science (PLoS)
• Subjects: Adenosine; Adult; Affinity chromatography; Alkaline phosphatase; Angiogenesis; Angiogenesis Inducing Agents - chemistry; Angiogenesis Inducing Agents - metabolism; Angiogenesis Inducing Agents - pharmacology; Animals; Biology; Capillary tubes; Cell culture; Cell migration; Cell Movement - drug effects; Cell proliferation; Chemistry; Chorioallantoic Membrane - drug effects; Chorioallantoic Membrane - embryology; Chromatography; Diabetes; Diabetic retinopathy; Embryo, Mammalian - metabolism; Embryos; Endothelial cells; Endothelial Cells - cytology; Endothelial Cells - drug effects; Endothelial Cells - enzymology; Endothelium; Endothelium, Vascular - cytology; Endothelium, Vascular - drug effects; Endothelium, Vascular - metabolism; Homogeneity; Humans; In Vitro Techniques; Incubation; Mammals; Mechanical stimuli; Medicine; Mitogen-Activated Protein Kinases - metabolism; Molecular Weight; Muscle, Smooth, Vascular - cytology; Muscles; Myocytes, Smooth Muscle - cytology; Myocytes, Smooth Muscle - drug effects; Myocytes, Smooth Muscle - metabolism; Nitric oxide; Phosphatase; Phosphorylation - drug effects; Physiology; Platelet-derived growth factor; Platelet-Derived Growth Factor - pharmacology; Rats; Rats, Wistar; Receptors; Secretion; Secretome; Shear stress; Shear stresses; Smooth muscle; Spectrometry; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Spheroids; Suramin; Tubes; Uracil Nucleotides - chemistry; Uracil Nucleotides - metabolism; Uracil Nucleotides - pharmacology; Vascular endothelial growth factor; Veins & arteries; Yolk; Yolk sac; Germany
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0068575

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 (1.34 ± 0.26 nmol L(-1)). In conclusion, Up4U is a novel strong human endothelium-derived angiogenic factor.