title: Angiogenic endothelium shows lactadherin-dependent phagocytosis of aged erythrocytes and apoptotic cells.
abstract: Angiogenic endothelium plays a crucial role in tumor growth. During angiogenesis, complex alterations in the microenvironment occur. In response, the endothelium undergoes phenotypic changes, for example overexpression of alpha(v)-integrins. Here, we show that the overexpression of alpha(v)-integrins on angiogenic endothelial cells is engaged in phagocytic actions involving binding ("tethering") and uptake ("tickling") of lactadherin (also termed MFG-E8)--opsonized particles. Phosphatidylserine (PS)--exposing multilamellar vesicles, "aged" erythrocytes, and apoptotic melanoma cells incubated with lactadherin were all phagocytosed by angiogenic endothelial cells in vitro. Furthermore, we demonstrated lactadherin expression in and around tumor blood vessels making opsonization in situ plausible. By engineering the surface of erythrocytes with covalently coupled cyclic Arg-Gly-Asp (RGD) peptides--mimicking lactadherin opsonization--we could induce phagocytosis by angiogenic endothelial cells both in vitro and in vivo. In vitro, this was confirmed by cytochalasin D preincubation. When RGD-erythrocytes were administered intravenously in tumor-bearing mice, blood vessel congestion followed by tumor core necrosis was seen. Moreover, RGD-erythrocytes could delay tumor growth in a murine melanoma model, possibly through induction of tumor infarctions. These results reveal that angiogenic endothelial cells have phagocytic properties for lactadherin-opsonized large particles and apoptotic cells. Implications of our findings for diagnostic and therapy of angiogenesis-driven diseases are discussed.
authors.affiliation: Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands.
authors.forename: Marcel H A M
authors.initials: MH
authors.lastname: Fens
authors.forename: Enrico
authors.initials: E
authors.lastname: Mastrobattista
authors.forename: Anko M
authors.initials: AM
authors.lastname: de Graaff
authors.forename: Frits M
authors.initials: FM
authors.lastname: Flesch
authors.forename: Anton
authors.initials: A
authors.lastname: Ultee
authors.forename: Jan T
authors.initials: JT
authors.lastname: Rasmussen
authors.forename: Grietje
authors.initials: G
authors.lastname: Molema
authors.forename: Gert
authors.initials: G
authors.lastname: Storm
authors.forename: Raymond M
authors.initials: RM
authors.lastname: Schiffelers
mesh_terms: D000818:Animals; D000954:Antigens, Surface / Q000502:physiology*; D017209:Apoptosis; D002417:Cattle; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000473:pathology / Q000502:physiology*; D004905:Erythrocyte Aging; D004912:Erythrocytes / Q000502:physiology*; D006801:Humans; D008546:Melanoma, Experimental / Q000473:pathology; D051379:Mice; D008894:Milk Proteins / Q000378:metabolism*; D009389:Neovascularization, Pathologic / Q000473:pathology*; D010587:Phagocytosis
chemical_list: D000954:Antigens, Surface; D008894:Milk Proteins