title: Characterization of the anti-angiogenic properties of arresten, an alpha1beta1 integrin-dependent collagen-derived tumor suppressor.
abstract: Physiological and pathological turnover of basement membranes liberates biologically active cryptic molecules. Several collagen-derived fragments possess anti-angiogenic activity. Arresten is the 26-kDa non-collagenous domain of type IV collagen alpha1 chain. It functions as an efficient inhibitor of angiogenesis and tumor growth in mouse models, but its anti-angiogenic mechanism is not completely known. Here we show that arresten significantly increases apoptosis of endothelial cells in vitro by decreasing the amount of anti-apoptotic molecules of the Bcl-family; Bcl-2 and Bcl-xL. Although the pro-apoptotic effect of arresten is endothelial cell specific in vitro, in mouse tumors arresten induced apoptosis both in endothelial and tumor cells. The tumor cell apoptosis is likely an indirect effect due to the inhibition of blood vessel growth into the tumor. The active site of arresten was localized by deletion mutagenesis within the C-terminal half of the molecule. We have previously shown that arresten binds to alpha1beta1 integrin on human umbilical vein endothelial cells. However, the microvascular endothelial cells (MLECs) are more important in the context of tumor vasculature. We show here that arresten binds also to the microvascular endothelial cells via alpha1beta1 integrin. Furthermore, it has no effect on Matrigel neovascularization or the viability of integrin alpha1 null MLECs. Tumors implanted on integrin alpha1 deficient mice show no integrin alpha1 expression in the host-derived vascular endothelium, and thus arresten does not inhibit the tumor growth. Collectively, this data sheds more light into the anti-angiogenic mechanism of arresten.
authors.affiliation: Center for Matrix Biology, Department of Medicine, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA, USA. pia.nyberg@oulu.fi
authors.forename: Pia
authors.initials: P
authors.lastname: Nyberg
authors.forename: Liang
authors.initials: L
authors.lastname: Xie
authors.forename: Hikaru
authors.initials: H
authors.lastname: Sugimoto
authors.forename: Pablo
authors.initials: P
authors.lastname: Colorado
authors.forename: Malin
authors.initials: M
authors.lastname: Sund
authors.forename: Kathryn
authors.initials: K
authors.lastname: Holthaus
authors.forename: Akulapalli
authors.initials: A
authors.lastname: Sudhakar
authors.forename: Tuula
authors.initials: T
authors.lastname: Salo
authors.forename: Raghu
authors.initials: R
authors.lastname: Kalluri
mesh_terms: D020533:Angiogenesis Inhibitors / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D001011:Aorta / Q000166:cytology; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D002417:Cattle; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002478:Cells, Cultured; D024141:Collagen Type IV / Q000737:chemistry* / Q000235:genetics; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects*; D006801:Humans; D039222:Integrin alpha1beta1 / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D016297:Mutagenesis, Site-Directed; D017434:Protein Structure, Tertiary / Q000502:physiology; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000187:drug effects / Q000378:metabolism; D011651:Pulmonary Artery / Q000166:cytology; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D051020:bcl-X Protein / Q000187:drug effects / Q000378:metabolism
chemical_list: D020533:Angiogenesis Inhibitors; D024141:Collagen Type IV; D039222:Integrin alpha1beta1; D019253:Proto-Oncogene Proteins c-bcl-2; D011994:Recombinant Proteins; D025521:Tumor Suppressor Proteins; D051020:bcl-X Protein
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