Abstract

Extracellular matrix (ECM) is a complex mixture of proteoglycans that function in cell adhesion, migration, growth and differentiation. In addition, the ECM serves as a reservoir for a number of growth factors, proteinases and proteinase inhibitors essential for ECM homeostasis. Recent new information from the applicant's laboratory has shown that one of the serine proteinase inhibitors found in the ECM is a 32kDa glycoprotein consisting of three tandemly-arranged Kunitz-type proteinase inhibitor domains homologous to tissue factor pathway inhibitor, an important negative regulator of the extrinsic pathway of blood coagulation. This novel inhibitor, designated as type-2 tissue factor pathway inhibitor, or TFPI-2, is a potent inhibitor of trypsin, plasmin, kallikrein and factor XIa amidolytic activity in the test tube. A variety of human endothelial cells synthesize TFPI-2, and 60-90 percent of the synthesized TFPI-2 is secreted by the endothelial cells into their ECM. Inflammatory mediators upregulate endothelial cell TFPI-2 synthesis 2-14 fold. In addition, evidence was obtained that recombinant TFPI-2 inhibited the plasmin-mediated invasion and degradation of Matrigel by a high invasive fibrosarcoma cell line in a dose-dependent manner. Furthermore, treatment of confluent endothelial cell monolayers with anti-TFPI-2 IgG detached the cells from the substrata in a time and IgG concentration dependent manner, suggesting an important role for TFPI-2 in maintaining the integrity of the ECM essential for cell attachment. In spite of these findings several questions concerning the physiological role of TFPI-2 remain unanswered, such as (1) which Kurutz domain is responsible for its inhibitory activity, (2) what proteinase(s) are regulated by ECM-bound TFPI-2, and (3) what are the regulatory mechanisms of TFPI-2 expression. The objectives of this proposal are to accomplish a comprehensive characterization of TFPI-2 in order to gain further insight into its biological role in normal and pathological ECM turnover.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL064119-03
Application #
6527289
Study Section
Hematology Subcommittee 2 (HEM)
Program Officer
Link, Rebecca P
Project Start
2000-09-05
Project End
2004-07-31
Budget Start
2002-08-01
Budget End
2003-07-31
Support Year
3
Fiscal Year
2002
Total Cost
$294,000
Indirect Cost

  Institution

Name
University of New Mexico
Department
Pathology
Type
Schools of Medicine
DUNS #
829868723
City
Albuquerque
State
NM
Country
United States
Zip Code
87131

  Publications

Kempaiah, Prakasha; Danielson, Leslie A; Barry, Marc et al. (2009) Comparative effects of aprotinin and human recombinant R24K KD1 on temporal renal function in Long-Evans rats. J Pharmacol Exp Ther 331:940-5
Kempaiah, Prakasha; Chand, Hitendra S; Kisiel, Walter (2009) Human tissue factor pathway inhibitor-2 is internalized by cells and translocated to the nucleus by the importin system. Arch Biochem Biophys 482:58-65
Kempaiah, Prakasha; Kisiel, Walter (2008) Human tissue factor pathway inhibitor-2 induces caspase-mediated apoptosis in a human fibrosarcoma cell line. Apoptosis 13:702-15
Shoji, Mamoru; Sun, Aiming; Kisiel, Walter et al. (2008) Targeting tissue factor-expressing tumor angiogenesis and tumors with EF24 conjugated to factor VIIa. J Drug Target 16:185-97
Sierko, Ewa; Wojtukiewicz, Marek Z; Kisiel, Walter (2007) The role of tissue factor pathway inhibitor-2 in cancer biology. Semin Thromb Hemost 33:653-9
Wojtukiewicz, Marek Z; Sierko, Ewa; Kisiel, Walter (2007) The role of hemostatic system inhibitors in malignancy. Semin Thromb Hemost 33:621-42
Chand, H S; Kisiel, W (2007) Quantitative real-time reverse transcription polymerase chain reaction analysis of a novel tissue factor splice variant in select human solid tumors. J Thromb Haemost 5:640-1
Kempaiah, Prakasha; Chand, Hitendra S; Kisiel, Walter (2007) Identification of a human TFPI-2 splice variant that is upregulated in human tumor tissues. Mol Cancer 6:20
Xu, Zhenhua; Maiti, Debasish; Kisiel, Walter et al. (2006) Tissue factor pathway inhibitor-2 is upregulated by vascular endothelial growth factor and suppresses growth factor-induced proliferation of endothelial cells. Arterioscler Thromb Vasc Biol 26:2819-25
Torres-Collado, Antoni X; Kisiel, Walter; Iruela-Arispe, Maria L et al. (2006) ADAMTS1 interacts with, cleaves, and modifies the extracellular location of the matrix inhibitor tissue factor pathway inhibitor-2. J Biol Chem 281:17827-37

Showing the most recent 10 out of 21 publications