) High molecular weight kininogen (HK) is an abundant plasma glycoprotein that plays a central role in contact activation. HK is cleaved to two-chain high molecular weight kininogen (HKa) on the endothelial cell surface. Cleavage of HK to HKa leads to the release of bradykinin, and is accompanied by a dramatic structural arrangement. Although binding of bradykinin to endothelial cells leads to well-defined responses, functional consequences that result from binding of HKa have not been reported. We have observed that HKa, but not HK, inhibits endothelial proliferation and induces endothelial apoptosis in a Zn2+-dependent manner. Proliferation is inhibited by 50% at an HKa concentration of 10 nM. In contrast, HKa does not inhibit the proliferation of aortic smooth muscle or trophoblast cells, or tumor-derived cell lines. In mice, HKa inhibits the neovascularization of basic FGF-containing Matrigel plugs. Modeling of HKa domain 5 (D5), which also induces endothelial cell apoptosis in a Zn2+-dependent manner, reveals that the structure of the heparin and zinc-binding pharmacophores within this domain closely resemble those within the antiangiogenic polypeptide, endostatin. We have also identified peptides from the endothelial cell binding regions of HKa domains 3 and 5 that inhibit endothelial cell proliferation at low micromolar to nanomolar concentrations; D5, but not D3-derived peptide exhibit Zn2+ dependence. Further insight into the mechanism of HKa is provided by the observations that HKa causes apoptosis of only subconfluent, proliferative endothelial cells, and that culture of endothelial cells on collagen I, IV or a smooth muscle cell-derived matrix protects against HKa-induced apoptosis. In this application, we propose to further define the mechanism of Hka-induced endothelial cell apoptosis, and to use site-directed mutagenesis to determine whether the Zn2+-binding pharmacophore(s) in domain 5 are important in antiangiogenic activity. We will also characterize the in viva antiangiogenic activity of HKa domains 3 and 5, as well as peptides derived from them. Finally, we propose to identify the endothelial cell receptor that mediate the antiangiogenic effects of Hka. These studies should provide new information concerning the physiologic regulation of angiogenesis, and may lead to identification of new antiangiogenic agents.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
1R01CA083134-01A1
Application #
6130885
Study Section
Special Emphasis Panel (ZCA1-SRRB-3 (J1))
Program Officer
Wolpert, Mary K
Project Start
2000-06-01
Project End
2005-05-31
Budget Start
2000-06-01
Budget End
2001-05-31
Support Year
1
Fiscal Year
2000
Total Cost
$360,861
Indirect Cost
Name
Case Western Reserve University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
077758407
City
Cleveland
State
OH
Country
United States
Zip Code
44106
Schmaier, A H; McCrae, K R (2007) The plasma kallikrein-kinin system: its evolution from contact activation. J Thromb Haemost 5:2323-9
Sun, Danyu; McCrae, Keith R (2006) Endothelial-cell apoptosis induced by cleaved high-molecular-weight kininogen (HKa) is matrix dependent and requires the generation of reactive oxygen species. Blood 107:4714-20
An, Feng-Qi; Folarin, Hope Merlene; Compitello, Nicole et al. (2006) Long-term-infected telomerase-immortalized endothelial cells: a model for Kaposi's sarcoma-associated herpesvirus latency in vitro and in vivo. J Virol 80:4833-46
McCrae, Keith R; Donate, Fernando; Merkulov, Sergei et al. (2005) Inhibition of angiogenesis by cleaved high molecular weight kininogen (HKa) and HKa domain 5. Curr Cancer Drug Targets 5:519-28
Juarez, Jose C; Guan, Xiaojun; Shipulina, Natalya V et al. (2002) Histidine-proline-rich glycoprotein has potent antiangiogenic activity mediated through the histidine-proline-rich domain. Cancer Res 62:5344-50
Zhang, Jing-Chuan; Qi, Xiaoping; Juarez, Jose et al. (2002) Inhibition of angiogenesis by two-chain high molecular weight kininogen (HKa) and kininogen-derived polypeptides. Can J Physiol Pharmacol 80:85-90
Dowlati, Afshin; Robertson, Kelly; Cooney, Matthew et al. (2002) A phase I pharmacokinetic and translational study of the novel vascular targeting agent combretastatin a-4 phosphate on a single-dose intravenous schedule in patients with advanced cancer. Cancer Res 62:3408-16
Zhang, Jing-Chuan; Donate, Fernando; Qi, Xiaoping et al. (2002) The antiangiogenic activity of cleaved high molecular weight kininogen is mediated through binding to endothelial cell tropomyosin. Proc Natl Acad Sci U S A 99:12224-9
Zhang, J C; Claffey, K; Sakthivel, R et al. (2000) Two-chain high molecular weight kininogen induces endothelial cell apoptosis and inhibits angiogenesis: partial activity within domain 5. FASEB J 14:2589-600