title: Collagenolysis-dependent angiogenesis mediated by matrix metalloproteinase-13 (collagenase-3).
abstract: We have demonstrated previously that new blood vessel formation induced by angiogenic growth factors in onplants placed on the chorioallantoic membrane (CAM) of the chick embryos is critically dependent on the cleavage of fibrillar collagen by a previously unidentified interstitial collagenase. In the present study we have used a quantitative CAM angiogenesis system to search for and functionally characterize host avian collagenases responsible for the collagen remodeling associated with angiogenesis. Among the matrix metalloproteinases (MMPs) identified in the CAM onplant tissue, the chicken MMP-13 (chMMP-13) was the only enzyme whose induction and expression coincided with the onset of angiogenesis and blood vessel formation. The chMMP-13 cDNA has been cloned and recombinantly expressed. The chMMP-13 protein has been purified, characterized in vitro, and examined in situ in the CAM. MMP-13-positive cells appear in the CAM shortly after angiogenic stimulation and then accumulate in the collagen onplant tissue. Morphologically, the chMMP-13-containing cells appear as hematopoietic cells of monocyte/macrophage lineage. In vitro, the chMMP-13 proenzyme is rapidly and efficiently activated through the urokinase plasminogen activator/plasminogen/plasmin cascade into a collagenase capable of cleaving native but not the (r/r) mutant collagenase-resistant collagen. Surprisingly, nanogram levels of purified chMMP-13 elicit an angiogenic response in the CAM onplants comparable with that induced by the angiogenic growth factors. The chMMP-13-mediated response was efficiently blocked by select protease inhibitors indicating that plasmin-activated chMMP-13 can function as an angiogenic factor in vivo. Altogether, the results of this study extend the physiological role of MMP-13, previously associated with cartilage/bone resorption, to the collagen remodeling involved in the angiogenic cascade.
authors.affiliation: Department of Cell Biology, The Scripps Research Institute, La Jolla, California 92037, USA.
authors.forename: Andries
authors.initials: A
authors.lastname: Zijlstra
authors.forename: Ronald T
authors.initials: RT
authors.lastname: Aimes
authors.forename: Dan
authors.initials: D
authors.lastname: Zhu
authors.forename: Karine
authors.initials: K
authors.lastname: Regazzoni
authors.forename: Tatyana
authors.initials: T
authors.lastname: Kupriyanova
authors.forename: Marco
authors.initials: M
authors.lastname: Seandel
authors.forename: Elena I
authors.initials: EI
authors.lastname: Deryugina
authors.forename: James P
authors.initials: JP
authors.lastname: Quigley
mesh_terms: D000482:Allantois / Q000098:blood supply / Q000378:metabolism / Q000648:ultrastructure; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002642:Chick Embryo; D002823:Chorion / Q000098:blood supply / Q000378:metabolism / Q000648:ultrastructure; D003094:Collagen / Q000235:genetics / Q000378:metabolism*; D017364:Collagenases / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000096:biosynthesis / Q000235:genetics; D005346:Fibroblast Growth Factors / Q000494:pharmacology; D053509:Matrix Metalloproteinase 13; D051379:Mice; D008969:Molecular Sequence Data; D018919:Neovascularization, Physiologic / Q000502:physiology*; D010958:Plasminogen / Q000494:pharmacology; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000378:metabolism; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology
chemical_list: D004792:Enzyme Precursors; D011994:Recombinant Proteins; D042461:Vascular Endothelial Growth Factor A; D019716:Tissue Inhibitor of Metalloproteinase-2; D005346:Fibroblast Growth Factors; D010958:Plasminogen; D003094:Collagen; D014568:Urokinase-Type Plasminogen Activator; D017364:Collagenases; D053509:Matrix Metalloproteinase 13; C507081:Mmp13 protein, mouse