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(Arteriosclerosis, Thrombosis, and Vascular Biology. 2007;27:310.)
© 2007 American Heart Association, Inc.

Vascular Biology

Adenovirus-Mediated Expression of Tissue Factor Pathway Inhibitor-2 Inhibits Endothelial Cell Migration and Angiogenesis

Lacramioara Ivanciu; Robert D. Gerard; Haiwang Tang; Florea Lupu; Cristina Lupu

From the Cardiovascular Biology Research Program (L.I., H.T., F.L., C.L.), Oklahoma Medical Research Foundation, Oklahoma City; the Department of Internal Medicine (R.D.G.), University of Texas Southwestern Medical Center, Dallas; and the Department of Pathology (F.L.), Oklahoma University Health Sciences Center, Oklahoma City.

Correspondence to Cristina Lupu, PhD, Oklahoma Medical Research Foundation, 825 NE 13th Street, Oklahoma City, OK 73104. E-mail cristina-lupu{at}omrf.ouhsc.edu

Objective— Extracellular matrix (ECM) remodeling during angiogenesis is accomplished through plasmin-dependent pericellular proteolysis and through the action of matrix metalloproteinases (MMPs). Because tissue factor pathway inhibitor-2 (TFPI-2), a Kunitz-type protease inhibitor with prominent ECM localization, inhibits plasmin and MMPs activity, we investigated the role of TFPI-2 in endothelial cell (EC) migration and angiogenesis.

Methods and Results— Real-time polymerase chain reaction and immunostaining showed that the expression of TFPI-2 mRNA and protein was upregulated in migrating ECs. The effect of TFPI-2 on angiogenesis was studied in mouse models of Matrigel and polyvinylalcohol sponge implants by overexpressing TFPI-2 through infection with a replication-deficient adenovirus (AdTFPI-2). Using (immuno)fluorescence and confocal microscopy we observed that TFPI-2 reduced neovascularization and promoted ECM deposition. Lateral cell migration and capillary tube formation in vitro also were impaired by TFPI-2, a process reversed by anti–TFPI-2 antibodies. Increased apoptosis occurred both in AdTFPI-2–treated ECs and in the mouse implants. Zymography and assays in the absence of plasminogen confirmed plasmin inhibition as a main mechanism through which TFPI-2 inhibits EC migration.

Conclusions— Our data suggest that TFPI-2 may be an important regulator of aberrant angiogenesis associated with tumor growth/metastasis, cardiovascular diseases, chronic inflammation, or diabetes.

This study shows that overexpression of TFPI-2 is an effective strategy to suppress angiogenesis. The mechanism apparently involves inhibition by TFPI-2 of plasmin and/or MMPs, thus reducing cell migration, promoting extracellular matrix deposition, and inhibiting neovascularization. The present data raise the possibility that TFPI-2 is a negative regulator of physiological angiogenesis.

Key Words: TFPI-2 • angiogenesis • endothelial cell • adenovirus gene delivery • mouse model