title: Nerve growth factor-induced migration of endothelial cells.
abstract: Nerve growth factor (NGF) is a well known neurotropic and neurotrophic agonist in the nervous system, which recently was shown to also induce angiogenic effects in endothelial cells (ECs). To measure NGF effects on the migration of cultured ECs, an important step in neoangiogenesis, we optimized an omnidirectional migration assay using human aortic endothelial cells (HAECs) and validated the assay with human recombinant basic fibroblast growth factor (rhbFGF) and human recombinant vascular endothelial growth factor (rhVEGF). The potencies of nerve growth factor purified from various species (viper, mouse, and recombinant human) to stimulate HAEC migration was similar to that of VEGF and basic fibroblast growth factor (bFGF) (EC50 of approximately 0.5 ng/ml). Recombinant human bFGF was significantly more efficacious than either viper NGF or rhVEGF, both of which stimulated HAEC migration by approximately 30% over basal spontaneous migration. NGF-mediated stimulation of HAEC migration was completely blocked by the NGF/TrkA receptor antagonist K252a [(8R*,9S*,11S*)-(/)-9-hydroxy-9-methoxycarbonyl-8-methyl-2,3,9,10-tetrahydro-8,11-epoxy-1H,-8H,11H-2,7b,11a-triazadibenzo(a,g)cycloocta(c,d,e)trindene-1-one] (30 nM) but not by the VEGF/Flk receptor antagonist SU-5416 [3-[(2,4-dimethylpyrrol-5-yl) methylidenyl]-indolin-2-one] (250 nM), indicating a direct effect of NGF via TrkA receptor activation on HAEC migration. Viper NGF stimulation of HAEC migration was additively increased by either rhVEGF or rhbFGF, suggesting a potentiating interaction between their tyrosine kinase receptor signaling pathways. Viper NGF represents a novel pharmacological tool to investigate possible TrkA receptor subtypes in endothelial cells. The ability of NGF to stimulate migration of HAEC cells in vitro implies that this factor may play an important role in the cardiovascular system besides its well known effects in the nervous system.
authors.affiliation: Calhoun Chair Professor of Cellular Tissue Engineering, School of Biomedical Engineering, Science and Health Systems, Drexel University, 3141 Chestnut St., Philadelphia, PA 19104, USA.
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Dollé
authors.forename: Amir
authors.initials: A
authors.lastname: Rezvan
authors.forename: Fred D
authors.initials: FD
authors.lastname: Allen
authors.forename: Philip
authors.initials: P
authors.lastname: Lazarovici
authors.forename: Peter I
authors.initials: PI
authors.lastname: Lelkes
mesh_terms: D000313:Adrenal Medulla / Q000166:cytology; D000818:Animals; D001011:Aorta / Q000166:cytology; D002227:Carbazoles / Q000494:pharmacology; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects*; D004730:Endothelium, Vascular / Q000166:cytology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D016222:Fibroblast Growth Factor 2 / Q000235:genetics / Q000494:pharmacology; D006801:Humans; D026121:Indole Alkaloids; D007211:Indoles / Q000494:pharmacology; D051379:Mice; D008954:Models, Biological; D020932:Nerve Growth Factor / Q000032:analysis / Q000235:genetics / Q000494:pharmacology*; D011758:Pyrroles / Q000494:pharmacology; D051381:Rats; D020800:Receptor, Nerve Growth Factor / Q000187:drug effects; D020917:Receptor, trkA / Q000037:antagonists & inhibitors; D011994:Recombinant Proteins / Q000494:pharmacology; D015203:Reproducibility of Results; D013997:Time Factors; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000494:pharmacology; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000037:antagonists & inhibitors; D017819:Viperidae
chemical_list: D002227:Carbazoles; D004791:Enzyme Inhibitors; D026121:Indole Alkaloids; D007211:Indoles; D011758:Pyrroles; D020800:Receptor, Nerve Growth Factor; D011994:Recombinant Proteins; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D016222:Fibroblast Growth Factor 2; C116890:Semaxinib; D020932:Nerve Growth Factor; C049985:staurosporine aglycone; D020917:Receptor, trkA; D040301:Vascular Endothelial Growth Factor Receptor-2