Abstract
The endothelial cell (EC)-derived tissue inhibitor of metalloproteinase-2 (TIMP-2) and pericyte-derived TIMP-3 are shown to coregulate human capillary tube stabilization following EC-pericyte interactions through a combined ability to block EC tube morphogenesis and regression in three-dimensional collagen matrices. EC-pericyte interactions strongly induce TIMP-3 expression by pericytes, whereas ECs produce TIMP-2 in EC-pericyte cocultures. Using small interfering RNA technology, the suppression of EC TIMP-2 and pericyte TIMP-3 expression leads to capillary tube regression in these cocultures in a matrix metalloproteinase-1 (MMP-1)-, MMP-10-, and ADAM-15 (a disintegrin and metalloproteinase-15)-dependent manner. Furthermore, we show that EC tube morphogenesis (lumen formation and invasion) is primarily controlled by the TIMP-2 and -3 target membrane type (MT) 1 MMP. Additional targets of these inhibitors include MT2-MMP and ADAM-15, which also regulate EC invasion. Mutagenesis experiments reveal that TIMP-3 requires its proteinase inhibitory function to induce tube stabilization. Overall, these data reveal a novel role for both TIMP-2 and -3 in the pericyte-induced stabilization of newly formed vascular networks that are predisposed to undergo regression and reveal specific molecular targets of the inhibitors regulating these events.
Publication types
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Research Support, N.I.H., Extramural
MeSH terms
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ADAM Proteins / genetics
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ADAM Proteins / metabolism
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Angiogenesis Inhibitors / genetics
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Angiogenesis Inhibitors / physiology*
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Animals
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Capillaries / cytology
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Capillaries / growth & development*
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Capillaries / metabolism
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Cattle
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Collagen
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Embryo, Mammalian / blood supply
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Embryo, Mammalian / metabolism
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Endothelium, Vascular / cytology
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Endothelium, Vascular / growth & development*
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Endothelium, Vascular / metabolism
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Gene Expression Regulation
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Humans
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Matrix Metalloproteinase 1 / metabolism
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Matrix Metalloproteinase 10 / metabolism
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Matrix Metalloproteinases, Membrane-Associated / genetics
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Matrix Metalloproteinases, Membrane-Associated / metabolism
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Membrane Proteins / genetics
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Membrane Proteins / metabolism
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Mice
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Models, Cardiovascular
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Mutagenesis
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Pericytes / metabolism*
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RNA Interference
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Tissue Inhibitor of Metalloproteinase-2 / antagonists & inhibitors
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Tissue Inhibitor of Metalloproteinase-2 / genetics
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Tissue Inhibitor of Metalloproteinase-2 / physiology*
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Tissue Inhibitor of Metalloproteinase-3 / antagonists & inhibitors
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Tissue Inhibitor of Metalloproteinase-3 / genetics
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Tissue Inhibitor of Metalloproteinase-3 / physiology*
Substances
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Angiogenesis Inhibitors
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Membrane Proteins
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TIMP3 protein, human
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Tissue Inhibitor of Metalloproteinase-3
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Tissue Inhibitor of Metalloproteinase-2
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Collagen
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ADAM Proteins
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ADAM15 protein, human
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Matrix Metalloproteinases, Membrane-Associated
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Matrix Metalloproteinase 10
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Matrix Metalloproteinase 1