title: SILAC-based proteomics of human primary endothelial cell morphogenesis unveils tumor angiogenic markers.
abstract: Proteomics has been successfully used for cell culture on dishes, but more complex cellular systems have proven to be challenging and so far poorly approached with proteomics. Because of the complexity of the angiogenic program, we still do not have a complete understanding of the molecular mechanisms involved in this process, and there have been no in depth quantitative proteomic studies. Plating endothelial cells on matrigel recapitulates aspects of vessel growth, and here we investigate this mechanism by using a spike-in SILAC quantitative proteomic approach. By comparing proteomic changes in primary human endothelial cells morphogenesis on matrigel to general adhesion mechanisms in cells spreading on culture dish, we pinpoint pathways and proteins modulated by endothelial cells. The cell-extracellular matrix adhesion proteome depends on the adhesion substrate, and a detailed proteomic profile of the extracellular matrix secreted by endothelial cells identified CLEC14A as a matrix component, which binds to MMRN2. We verify deregulated levels of these proteins during tumor angiogenesis in models of multistage carcinogenesis. This is the most in depth quantitative proteomic study of endothelial cell morphogenesis, which shows the potential of applying high accuracy quantitative proteomics to in vitro models of vessel growth to shed new light on mechanisms that accompany pathological angiogenesis. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium with the data set identifier PXD000359.
authors: Department of Proteomics and Signal Transduction, Max-Planck Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany;
authors: Sara
authors: S
authors: Zanivan
authors: Federica
authors: F
authors: Maione
authors: Marco Y
authors: MY
authors: Hein
authors: Juan Ramon
authors: JR
authors: Hernández-Fernaud
authors: Pawel
authors: P
authors: Ostasiewicz
authors: Enrico
authors: E
authors: Giraudo
authors: Matthias
authors: M
authors: Mann
mesh_terms: D000818:Animals; D000954:Antigens, Surface / Q000235:genetics* / Q000378:metabolism; D014408:Biomarkers, Tumor / Q000235:genetics* / Q000378:metabolism; D002247:Carbon Isotopes; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000235:genetics* / Q000378:metabolism; D002454:Cell Differentiation; D002471:Cell Transformation, Neoplastic / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D003094:Collagen / Q000737:chemistry; D004338:Drug Combinations; D005109:Extracellular Matrix / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D007553:Isotope Labeling; D007797:Laminin / Q000737:chemistry; D037181:Lectins, C-Type / Q000235:genetics* / Q000378:metabolism; D013058:Mass Spectrometry; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D009024:Morphogenesis / Q000235:genetics; D009389:Neovascularization, Pathologic; D061251:Primary Cell Culture; D011485:Protein Binding; D011509:Proteoglycans / Q000737:chemistry; D040901:Proteomics; D015398:Signal Transduction
chemical_list: D000954:Antigens, Surface; D014408:Biomarkers, Tumor; C570898:CLEC14A protein, human; D002247:Carbon Isotopes; D015815:Cell Adhesion Molecules; D004338:Drug Combinations; C089826:EMILIN3 protein, human; D007797:Laminin; D037181:Lectins, C-Type; D008562:Membrane Glycoproteins; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen
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