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Abstracts

suggest a direct interaction of integrin 21 and SHP1, and indicate that SHP1 is an essential mediator of endorepellin antiangiogenic activity. doi:10.1016/j.matbio.2008.09.226

matrilysin and SDC1 repaired faster than cells co-expressing matrilysin and a cleavage-resistant SDC1. Consistent with these findings, injured SDC1deficient airway epithelium repaired faster than wild-type epithelium in vitro and in vivo. Additionally, SDC1 knockdown increased migration and decreased adhesion of BEAS-2B cells compared to control cells. Moreover, 1 integrin activity was diminished in SDC1-deficient conditions. Conclusion These data show that matrilysin sheds SDC1 during airway reepithelialization. Moreover, SDC1 shedding removes restrictions to migration by reducing integrin-mediated adhesion. Because wound edge epithelium can control acute inflammation and re-epithelialization, a dual role for matrilysin shedding of SDC1 may reflect co-opting of one process into others. doi:10.1016/j.matbio.2008.09.228

12 Hypertriglyceridemia caused by mutation of the basement membrane proteoglycan Type XVIII collagen Jeffrey D. Esko, Joseph R. Bishop, Maria Rita Passos-Bueno, Kristin I. Stanford, Erika Yeh, Joseph L. Witztum, Andre Bensadoun, Karen S. Moulton Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, California 92093, United States The heparan sulfate proteoglycan Type XVIII collagen (Col18) is found ubiquitously in the basements membranes of the vasculature in vertebrates. Mice deficient in Col18 (Col18a1/) are viable and fertile, but have broadened basement membranes, abnormal retinal vessel development and poor anchoring of vitreal collagen fibrils to the inner lining membrane of the retina. Similar retinal defects occur in subjects with Knobloch syndrome, a rare disorder characterized by myopia, vitreoretinal degeneration and retinal detachment attributed to mutations in Col18. Examination of Col18a1/ mice demonstrates fasting and post-prandial hypertriglyceridemia characterized by large triglyceride-rich lipoproteins. These lipoproteins likely derive from chylomicrons based on size and apolipoprotein composition. Col18a1/ mice exhibit delayed hydrolysis of triglycerides in the peripheral circulation due to inadequate access of particles to lipoprotein lipase, which is normally bound to receptors on the luminal side of the endothelium but deficient in mutant mice. Furthermore, patients with Knobloch Syndrome also have hypertriglyceridemia and accumulate triglyceride-rich lipoproteins under fasting conditions, a previously unrecognized consequence of the loss of Col18 in humans. These findings suggest that Col18 is required for transport or presentation of lipoprotein lipase in the vasculature and that alterations in its expression can lead to hypertriglyceridemia in mice and humans. doi:10.1016/j.matbio.2008.09.227

14 Synstatin, a Sdc1 peptide, blocks v3-dependent angiogenesis DeannaLee M. Beauvais, Brian J. Ell, Andrea R. McWhorter, Alan C. Rapraeger Department of Pathology and Laboratory Medicine, University of WisconsinMadison, Madison, WI 53706, United States Syndecan-1 (Sdc1), via an interaction site in its ectodomain, forms a ternary complex with the v3 integrin and the insulin-like growth factor-1 receptor (IGF1R). Formation of this complex is required for v3-dependent cell adhesion and invasion in human mammary carcinoma and endothelial cells. Thus, Sdc1's regulation of v3 integrin activity likely affects tumor metastasis and tumorinduced angiogenesis. To study this, we derive a novel peptide called synstatin, which contains the integrin regulatory site identified in the Sdc1 ectodomain. We show that synstatin (SSTN; IC50 0.3 M) competitively dissociates the Sdc1integrin-IGF1R complex, thus blocking integrin activation, as evidenced by a loss in WOW1 and/or soluble fibrinogen binding (v3 integrin activation sensors) and a block in cell adhesion, spreading and migration on vitronectin, an v3specific ligand. SSTN effectively blocks angiogenesis in vitro (i.e., 3D aortic ring assays) and in vivo when delivered systemically in mouse corneal-pocket angiogenesis assays. Moreover, using human mammary carcinoma cells subcutaneously implanted in nude mice, SSTN also inhibits tumor-induced angiogenesis a prerequisite step for tumor growth and metastatic dispersion. Thus, Sdc1 plays a critical regulatory role during angiogenesis that can be potently targeted by the SSTN peptide. Disruption of this regulatory mechanism holds promise as an anti-cancer therapy. doi:10.1016/j.matbio.2008.09.229

13 Syndecan-1 shedding facilitates airway re-epithelialization Peter Chen, Laura E. Abacherli, Ying Wang, Qinglang Li, Bill C. Parks Center for Lung Biology, University of Washington, Seattle, WA 98109, United States Introduction Matrilysin (MMP-7) is essential for airway re-epithelialization. Matrilysin also functions to regulate lung inflammatory cell recruitment through the cleavage of syndecan-1 (SDC1). This study was to determine if matrilysin sheds SDC1 to facilitate re-epithelialization. Methods Primary mouse tracheal epithelial cells were cultured at an airliquid interface and injured as an in vitro model of re-epithelialization. Mice were also injured with naphthalene to study re-epithelialization in vivo. Cell lines were also retroviral transduced to stably express either a) wild-type or an uncleavable SDC1 or b) SDC1 shRNA. Results Wounded wild-type epithelium shed SDC1 resulting in diminished SDC1 immunofluorescent signal at the wound front in vitro and in vivo. In contrast, SDC1 shedding was minimal, and SDC1 signal persisted in wounded matrilysin-deficient epithelium. Wounded COS-7 cells that stably co-express

15 Decorin binds to and downregulates the MET receptor Silvia Goldonia, Ashley Humphriesa, Rick T. Owensb, David J. McQuillanb, Renato V. Iozzoa a Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, Philadelphia, PA, United States b LifeCell Corporation, Branchburg, NJ, United States Decorin, a member of the small leucine-rich proteoglycan gene family, regulates tumor cell growth primarily by downregulating epidermal growth factor receptor (EGFR) activity and by interacting with other members of the ErbB family of receptor tyrosine kinase. Due to the complex binding repertoire of decorin towards multiple targets, we predicted a role for decorin in modulating the bioactivity of other tyrosine kinase receptors. We discovered that decorin binds to Met, the receptor for hepatocyte growth factor/scatter factor, leading to transient activation at the catalytic domain, followed by a rapid (T1/26 min) downregulation of total Met. This activity was independent of EGFR. Decorin induced Met suppression by a dual action of recruiting the ubiquitin ligase c-Cbl followed by enhanced proteasome degradation and by evoking shedding of the receptor ectodomain. In addition