title: Endorepellin evokes autophagy in endothelial cells.
abstract: Endorepellin, the C-terminal fragment of the heparan sulfate proteoglycan perlecan, possesses angiostatic activity via dual receptor antagonism, through concurrent binding to the α2β1 integrin and vascular endothelial growth factor receptor 2 (VEGFR2). Here, we discovered that soluble endorepellin induced autophagy in endothelial cells by modulating the expression of Beclin 1, LC3, and p62, three established autophagic markers. Moreover, endorepellin evoked expression of the imprinted tumor suppressor gene Peg3 and its co-localization with Beclin 1 and LC3 in autophagosomes, suggesting a major role for this gene in endothelial cell autophagy. Mechanistically, endorepellin induced autophagy by down-regulating VEGFR2 via the two LG1/2 domains, whereas the C-terminal LG3 domain, the portion responsible for binding the α2β1 integrin, was ineffective. Endorepellin also induced transcriptional activity of the BECN1 promoter in endothelial cells, and the VEGFR2-specific tyrosine kinase inhibitor, SU5416, blocked this effect. Finally, we found a correlation between endorepellin-evoked inhibition of capillary morphogenesis and enhanced autophagy. Thus, we have identified a new role for this endogenous angiostatic fragment in inducing autophagy through a VEGFR2-dependent but α2β1 integrin-independent pathway. This novel mechanism specifically targets endothelial cells and could represent a promising new strategy to potentiate the angiostatic effect of endorepellin and perhaps other angiostatic matrix proteins.
authors.affiliation: From the Department of Pathology, Anatomy, and Cell Biology and the Cancer Cell Biology and Signaling Program, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania 19107.
authors.forename: Chiara
authors.initials: C
authors.lastname: Poluzzi
authors.affiliation: From the Department of Pathology, Anatomy, and Cell Biology and the Cancer Cell Biology and Signaling Program, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania 19107.
authors.forename: Joshua
authors.initials: J
authors.lastname: Casulli
authors.affiliation: From the Department of Pathology, Anatomy, and Cell Biology and the Cancer Cell Biology and Signaling Program, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania 19107.
authors.forename: Atul
authors.initials: A
authors.lastname: Goyal
authors.affiliation: From the Department of Pathology, Anatomy, and Cell Biology and the Cancer Cell Biology and Signaling Program, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania 19107.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Mercer
authors.affiliation: From the Department of Pathology, Anatomy, and Cell Biology and the Cancer Cell Biology and Signaling Program, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania 19107.
authors.forename: Thomas
authors.initials: T
authors.lastname: Neill
authors.affiliation: From the Department of Pathology, Anatomy, and Cell Biology and the Cancer Cell Biology and Signaling Program, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania 19107 renato.iozzo@jefferson.edu.
authors.forename: Renato V
authors.initials: RV
authors.lastname: Iozzo
mesh_terms: D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D001343:Autophagy / Q000502:physiology*; D000071186:Beclin-1; D002478:Cells, Cultured; D058546:Class III Phosphatidylinositol 3-Kinases / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology* / Q000201:enzymology / Q000378:metabolism; D019812:Heparan Sulfate Proteoglycans / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D038982:Integrin alpha2beta1 / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D009024:Morphogenesis; D010446:Peptide Fragments / Q000378:metabolism / Q000502:physiology*; D060888:Real-Time Polymerase Chain Reaction; D015398:Signal Transduction; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism
chemical_list: D051017:Apoptosis Regulatory Proteins; C491997:BECN1 protein, human; D000071186:Beclin-1; D019812:Heparan Sulfate Proteoglycans; D038982:Integrin alpha2beta1; D008565:Membrane Proteins; D010446:Peptide Fragments; C071927:perlecan; D058546:Class III Phosphatidylinositol 3-Kinases; D040301:Vascular Endothelial Growth Factor Receptor-2
keywords: Angiogenesis; Autophagy; Endothelial Cell; Extracellular Matrix; Extracellular Matrix Proteins; Proteoglycan
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