Identification of a potent endothelium-derived angiogenic factor.
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.
Charité-Universitaetsmedizin Berlin, Medizinische Klinik IV (CBF), Berlin, Germany.
Vera
V
Jankowski
Markus
M
Tölle
Thi Nguyet Anh
TN
Tran
Markus
M
van der Giet
Mirjam
M
Schuchardt
Kerstin
K
Lehmann
Doreen
D
Janke
Burkhard
B
Flick
Alberto
A
Ortiz
Alberto Arduan
AA
Ortiz
Maria D
MD
Sanchez-Niño
Niño Maria Dolores
NM
Sanchez
Martin
M
Tepel
Walter
W
Zidek
Joachim
J
Jankowski
D000328:Adult; D043925:Angiogenesis Inducing Agents / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D000818:Animals; D002465:Cell Movement / Q000187:drug effects; D049033:Chorioallantoic Membrane / Q000187:drug effects / Q000196:embryology; D004622:Embryo, Mammalian / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D008970:Molecular Weight; D009131:Muscle, Smooth, Vascular / Q000166:cytology; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014500:Uracil Nucleotides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology
D043925:Angiogenesis Inducing Agents; D010982:Platelet-Derived Growth Factor; D014500:Uracil Nucleotides; D020928:Mitogen-Activated Protein Kinases