This Article Full Text Full Text (PDF) All Versions of this Article: 111/13/1627    most recent 01.CIR.0000160364.05405.B5v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Shi, C.-S. Articles by Wu, H.-L. Search for Related Content PubMed PubMed Citation Articles by Shi, C.-S. Articles by Wu, H.-L. Pubmed/NCBI databases Substance via MeSH Related Collections Angiogenesis

(Circulation. 2005;111:1627-1636.)
© 2005 American Heart Association, Inc.

Molecular Cardiology

Evidence of Human Thrombomodulin Domain as a Novel Angiogenic Factor

Chung-Sheng Shi, MS; Guey-Yueh Shi, PhD; Yi-Sheng Chang, MD; Huai-Song Han, MS; Cheng-Hsiang Kuo, BS; Che Liu, MS; Huey-Chun Huang, PhD; Yu-Jia Chang, PhD; Pin-Shern Chen, PhD; Hua-Lin Wu, PhD

From the Department of Biochemistry and Molecular Biology (G.-Y.S., C.-H.K., C.L., H.-C.H., Y.-J.C., P.-S.C., H.-L.W.), Institute of Basic Medical Sciences (C.-S.S., H.-S.H.), Department of Ophthalmology (Y.-S.C.), and Institute of Clinical Medicine (Y.-S.C.), College of Medicine, National Cheng Kung University, Tainan, Taiwan, Republic of China.

Correspondence to Hua-Lin Wu, PhD, Professor, Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung University, 1 University Rd, Tainan, Taiwan, 701, Republic of China. E-mail halnwu{at}mail.ncku.edu.tw

Received July 20, 2004; revision received December 20, 2004; accepted December 21, 2004.

Background— Thrombomodulin is an anticoagulant, endothelial-cell-membrane glycoprotein. A recombinant thrombomodulin domain containing 6 epidermal growth factor–like structures exhibits mitogenic activity. This study explored the novel angiogenic effects of the recombinant domain using in vitro and in vivo models.

Methods and Results— Human recombinant thrombomodulin containing 6 epidermal growth factor–like structures (TMD2) and TMD2 plus a serine and threonine-rich domain (TMD23) were prepared using the Pichia pastoris expression system. Combined with purified TMD2 or TMD23, thrombin effectively activated protein C. TMD23 had higher activity than TMD2 in stimulating DNA synthesis in cultured human umbilical vein endothelial cells. Additionally, TMD23 stimulated chemotactic motility and capillarylike tube formation in human umbilical vein endothelial cells, an effect mediated through phosphorylation of extracellular signal–regulated kinase 1/2 and p38 mitogen-activated protein kinase and the phosphatidylinositol-3 kinase/Akt/endothelial nitric oxide synthase pathway. TMD23 also stimulated endothelial cell expression of matrix metalloproteinases and plasminogen activators, which mediated extracellular proteolysis, leading to endothelial cell invasion and migration during angiogenesis. Furthermore, TMD23-containing implants in rat cornea induced ingrowth of new blood vessels from the limbus. With the murine angiogenesis assay, TMD23 not only induced neovascularization coinjected with Matrigel and heparin but also enhanced angiogenesis in Matrigel containing melanoma A2058 cells in nude mice.

Conclusions— The recombinant thrombomodulin domain TMD23 enhanced the angiogenic response in vitro and in vivo, suggesting that thrombomodulin fragments may play a role in the formation of new vessels. These findings may provide a new therapeutic option for treating ischemic diseases.

Key Words: angiogenesis • endothelium • glycoproteins

This article has been cited by other articles:

				K. Otsu, S. Das, S. D. Houser, S. K. Quadri, S. Bhattacharya, and J. Bhattacharya Concentration-dependent inhibition of angiogenesis by mesenchymal stem cells Blood, April 30, 2009; 113(18): 4197 - 4205. [Abstract] [Full Text] [PDF]

				I.-C. Lo, T.-M. Lin, L.-H. Chou, S.-L. Liu, L.-W. Wu, G.-Y. Shi, H.-L. Wu, and M. J. Jiang Ets-1 mediates platelet-derived growth factor-BB-induced thrombomodulin expression in human vascular smooth muscle cells Cardiovasc Res, March 1, 2009; 81(4): 771 - 779. [Abstract] [Full Text] [PDF]

				C.-S. Shi, G.-Y. Shi, S.-M. Hsiao, Y.-C. Kao, K.-L. Kuo, C.-Y. Ma, C.-H. Kuo, B.-I. Chang, C.-F. Chang, C.-H. Lin, et al. Lectin-like domain of thrombomodulin binds to its specific ligand Lewis Y antigen and neutralizes lipopolysaccharide-induced inflammatory response Blood, November 1, 2008; 112(9): 3661 - 3670. [Abstract] [Full Text] [PDF]

				C.-F. Lam, P.-J. Chang, Y.-S. Huang, Y.-H. Sung, C.-C. Huang, M.-W. Lin, Y.-C. Liu, and Y.-C. Tsai Prolonged Use of High-Dose Morphine Impairs Angiogenesis and Mobilization of Endothelial Progenitor Cells in Mice Anesth. Analg., August 1, 2008; 107(2): 686 - 692. [Abstract] [Full Text] [PDF]

				C. Cappuzzello, R. Melchionna, A. Mangoni, G. Tripodi, P. Ferrari, L. Torielli, D. Arcelli, M. Helmer-Citterich, G. Bianchi, M. C. Capogrossi, et al. Role of rat {alpha} adducin in angiogenesis: Null effect of the F316Y polymorphism Cardiovasc Res, August 1, 2007; 75(3): 608 - 617. [Abstract] [Full Text] [PDF]

				M. Presta Response to comment on "cutting edge: extracellular high mobility group box-1 protein is a proangiogenic cytokine". J. Immunol., April 15, 2006; 176(8): 4513 - 4513. [Full Text] [PDF]

				M. David-Dufilho, E. M.-V. Brussel, G. Topal, L. Walch, A. Brunet, and F. Rendu Endothelial Thrombomodulin Induces Ca2+ Signals and Nitric Oxide Synthesis through Epidermal Growth Factor Receptor Kinase and Calmodulin Kinase II J. Biol. Chem., October 28, 2005; 280(43): 35999 - 36006. [Abstract] [Full Text] [PDF]