title: Anti-human activin receptor-like kinase 1 (ALK1) antibody attenuates bone morphogenetic protein 9 (BMP9)-induced ALK1 signaling and interferes with endothelial cell sprouting.
abstract: Genetic and molecular studies suggest that activin receptor-like kinase 1 (ALK1), a transforming growth factor β (TGF-β) type I receptor, and endoglin, a TGF-β co-receptor, play an essential role in vascular development and pathological angiogenesis. Several agents that interfere with ALK1 and endoglin function are currently in clinical trials for antiangiogenic activity in cancer therapy. One of these agents, PF-03446962 (anti-hALK1 antibody), shows promising results in the clinic. However, its effects on endothelial cell function and mechanism of action are unclear. Here we demonstrate that anti-hALK1 antibody selectively recognizes human ALK1. The anti-hALK1 antibody interfered with bone morphogenetic protein 9 (BMP9)-induced signaling in endothelial cells. Consistent with this notion, anti-hALK1 antibody was found to compete highly efficiently with the binding of the ALK1 ligand BMP9 and TGF-β to ALK1. Moreover, it prevented BMP9-dependent recruitment of co-receptor endoglin into this angiogenesis-mediating signaling complex. In addition, we demonstrated that anti-hALK1 antibody inhibited endothelial cell sprouting but did not directly interfere with vascular endothelial growth factor (VEGF) signaling, VEGF-induced proliferation, and migration of endothelial cells. Finally, we demonstrated that BMP9 in serum is essential for endothelial sprouting and that anti-hALK1 antibody inhibits this potently. Our data suggest that both the VEGF/VEGF receptor and the BMP9/ALK1 pathways are essential for stimulating angiogenesis, and targeting both pathways simultaneously may be an attractive strategy to overcome resistance to antiangiogenesis therapy.
authors.affiliation: Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands. L.A.van_Meeteren@lumc.nl
authors.forename: Laurens A
authors.initials: LA
authors.lastname: van Meeteren
authors.forename: Midory
authors.initials: M
authors.lastname: Thorikay
authors.forename: Simon
authors.initials: S
authors.lastname: Bergqvist
authors.forename: Evangelia
authors.initials: E
authors.lastname: Pardali
authors.forename: Corrado Gallo
authors.initials: CG
authors.lastname: Stampino
authors.forename: Dana
authors.initials: D
authors.lastname: Hu-Lowe
authors.forename: Marie-José
authors.initials: MJ
authors.lastname: Goumans
authors.forename: Peter
authors.initials: P
authors.lastname: ten Dijke
mesh_terms: D030301:Activin Receptors, Type II / Q000276:immunology* / Q000378:metabolism; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D005434:Flow Cytometry; D055427:Growth Differentiation Factor 2 / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D015398:Signal Transduction / Q000502:physiology*
chemical_list: D055427:Growth Differentiation Factor 2; C425364:ACVRL1 protein, human; D030301:Activin Receptors, Type II