Correlation between circulating endothelial cell counts and plasma thrombomodulin levels as markers for endothelial damage

Michiel H. Strijbos; Chandra Rao; Paul I. M. Schmitz; Jaco Kraan; Cor H. Lamers; Stefan Sleijfer; Leon W. M. M. Terstappen; Jan W. Gratama

doi:10.1160/TH08-02-0122

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2008; 100(04): 642-647
DOI: 10.1160/TH08-02-0122

Endothelium and Vascular Development

Schattauer GmbH

Correlation between circulating endothelial cell counts and plasma thrombomodulin levels as markers for endothelial damage

Michiel H. Strijbos

¹Laboratory of Medical and Tumor Immunology, Department of Medical Oncology, Erasmus University Medical Center, Rotterdam, The Netherlands

,

Chandra Rao

²Immunicon Corporation, Huntingdon Valley, Pennsylvania, USA

,

Paul I. M. Schmitz

³Department of Trials and Statistics, Erasmus University Medical Center, Rotterdam, The Netherlands

,

Jaco Kraan

¹Laboratory of Medical and Tumor Immunology, Department of Medical Oncology, Erasmus University Medical Center, Rotterdam, The Netherlands

,

Cor H. Lamers

¹Laboratory of Medical and Tumor Immunology, Department of Medical Oncology, Erasmus University Medical Center, Rotterdam, The Netherlands

,

Stefan Sleijfer

¹Laboratory of Medical and Tumor Immunology, Department of Medical Oncology, Erasmus University Medical Center, Rotterdam, The Netherlands

,

Leon W. M. M. Terstappen

²Immunicon Corporation, Huntingdon Valley, Pennsylvania, USA

,

Jan W. Gratama

¹Laboratory of Medical and Tumor Immunology, Department of Medical Oncology, Erasmus University Medical Center, Rotterdam, The Netherlands

› Author Affiliations

Abstract

PDF Download

Summary

Increased numbers of circulating endothelial cells (CEC) in peripheral blood have been observed in diseases with vascular involvement, and are considered a promising surrogate marker for vascular damage. It was the objective of this study to evaluate the correlation between putative soluble markers of endothelial injury, activation, and endothelial proliferation, and absolute numbers of CEC. CEC were evaluated in 125 healthy donors and 40 patients with metastatic carcinoma by automated CD146 driven immunomagnetic isolation. Plasma concentrations of E-selectin, endoglin, and thrombomodulin were assessed by ELISA in plasma obtained from 40 healthy donors and 40 patients. CEC numbers in blood were positively correlated with plasma thrombomodulin levels, but not with levels of E-se-lectin and endoglin. Multivariate analysis demonstrated a significant increase in CEC numbers with age. The levels of plasma biomarkers were not influenced by age. Higher levels of thrombomodulin and E-selectin were observed in males when compared to females. In conclusion, CEC numbers correlate positively with plasma levels of thrombomodulin.

Keywords

Biomarkers - CEC - endothelium - thrombomodulin - vascular damage

PDF (323 kb)

References

References
1 Folkman J. Tumor angiogenesis: therapeutic implications. New Engl J Med 1971; 285: 1182-1186.
2 Escudier B, Eisen T, Stadler WM. et al. Sorafenib in advanced clear-cell renal-cell carcinoma. New Engl J Med 2007; 356: 125-134.
3 Motzer RJ, Hutson TE, Tomczak P. et al. Sunitinib versus interferon alfa in metastatic renal-cell carcinoma. New Engl J Med 2007; 356: 115-124.
4 Hurwitz H, Fehrenbacher L, Novotny W. et al. Bevacizumab plus irinotecan, fluorouracil, and leucovorin for metastatic colorectal cancer. New Engl J Med 2004; 350: 2335-2342.
5 Ellis LM. Mechanisms of action of bevacizumab as a component of therapy for metastatic colorectal cancer. Sem Oncol 2006; 33 (05) (Suppl. 10) S1-7.
6 Curran SD, Muellner AU, Schwartz LH. Imaging response assessment in oncology. Cancer Imag 2006; 06: S126-130.
7 Cao Y, Liu Q. Therapeutic targets of multiple angiogenic factors for the treatment of cancer and metastasis. Adv Cancer Res 2007; 97: 203-224.
8 di Tomaso E, Capen D, Haskell A. et al. Mosaic tumor vessels: cellular basis and ultrastructure of focal regions lacking endothelial cell markers. Cancer Res 2005; 65: 5740-5749.
9 Blann AD, Woywodt A, Bertolini F. et al. Circulating endothelial cells. Biomarker of vascular disease. Thromb Haemost 2005; 93: 228-235.
10 Woywodt A, Blann AD, Kirsch T. et al. Isolation and enumeration of circulating endothelial cells by immunomagnetic isolation: proposal of a definition and a consensus protocol. J Thromb Haemost 2006; 04: 671-677.
11 Mancuso P, Burlini A, Pruneri G. et al. Resting and activated endothelial cells are increased in the peripheral blood of cancer patients. Blood 2001; 97: 3658-3661.
12 Duda DG, Cohen KS, Scadden DT. et al. A protocol for phenotypic detection and enumeration of circulating endothelial cells and circulating progenitor cells in human blood. Nat Protoc 2007; 02: 805-810.
13 Strijbos MH, Kraan J, den Bakker MA. et al. Cells meeting our immunophenotypic criteria of endothelial cells are large platelets. Cytometry B Clin Cytom 2007; 72: 86-93.
14 Rowand JL, Martin G, Doyle GV. et al. Endothelial cells in peripheral blood of healthy subjects and patients with metastatic carcinomas. Cytometry A 2007; 71: 105-113.
15 Smirnov DA, Foulk BW, Doyle GV. et al. Global gene expression profiling of circulating endothelial cells in patients with metastatic carcinomas. Cancer Res 2006; 66: 2918-2922.
16 Hrachovinova I, Cambien B, Hafezi-Moghadam A. et al. Interaction of P-selectin and PSGL-1 generates microparticles that correct hemostasis in a mouse model of hemophilia A. Nature Med 2003; 09: 1020-1025.
17 McEver RP. Adhesive interactions of leukocytes, platelets, and the vessel wall during hemostasis and inflammation. Thromb Haemost 2001; 86: 746-756.
18 Rini BI, Rathmell WK. Biological aspects and binding strategies of vascular endothelial growth factor in renal cell carcinoma. Clin Cancer Res 2007; 13: 741s-746s.
19 Zoppini G, Targher G, Zamboni C. et al. Effects of moderate-intensity exercise training on plasma biomarkers of inflammation and endothelial dysfunction in older patients with type 2 diabetes. Nutr Metab Cardiovasc Dis 2006; 16: 543-549.
20 Boffa MC, Karmochkine M. Thrombomodulin: an overview and potential implications in vascular disorders. Lupus 1998; 07 (Suppl. 02) S120-125.
21 Ishii H, Uchiyama H, Kazama M. Soluble thrombomodulin antigen in conditioned medium is increased by damage of endothelial cells. Thromb´Haemost 1991; 65: 618-623.
22 Kozuka K, Kohriyama T, Nomura E. et al. Endothelial markers and adhesion molecules in acute ischemic stroke--sequential change and differences in stroke subtype. Atherosclerosis 2002; 161: 161-168.
23 Bardin N, George F, Mutin M. et al. S-Endo 1, a pan-endothelial monoclonal antibody recognizing a novel human endothelial antigen. Tissue Antigens 1996; 48: 531-539.
24 Elshal MF, Khan SS, Takahashi Y. et al. CD146 (Mel-CAM), an adhesion marker of endothelial cells, is a novel marker of lymphocyte subset activation in normal peripheral blood. Blood 2005; 106: 2923-2924.
25 George F, Poncelet P, Laurent JC. et al. Cytofluorometric detection of human endothelial cells in whole blood using S-Endo 1 monoclonal antibody. J Immunol Methods 1991; 139: 65-75.
26 Gougos A, St Jacques S, Greaves A. et al. Identification of distinct epitopes of endoglin, an RGD-containing glycoprotein of endothelial cells, leukemic cells, and syncytiotrophoblasts. Int Immunol 1992; 04: 83-92.
27 Khan SS, Solomon MA, McCoy Jr JP. Detection of circulating endothelial cells and endothelial progenitor cells by flow cytometry. Cytometry B Clin Cytom 2005; 64: 1-8.
28 Widemann A, Sabatier F, Arnaud L. et al. CD146-based immunomagnetic enrichment followed by multi-parameter flow cytometry: a new approach to count circulating endothelial cells. J Thromb Haemost. 2008 prepub online.
29 Alvarez DF, Huang L, King JA. et al. Lung microvascular endothelium is enriched with progenitor cells that exhibit vasculogenic capacity. Am J Physiol Lung Cell Mol Physiol 2008; 294: L419-430.
30 Rustom R, Leggat H, Tomura HR. et al. Plasma thrombomodulin in renal disease: effects of renal function and proteinuria. Clin Nephrol 1998; 50: 337-341.
31 Xu GB, Luo LH, Lu XG. Detection of thrombomodulin in both plasma and tissue extracts of cancer patients and its clinical significance. Zhejiang Da Xue Xue Bao Yi Xue Ban 2003; 32: 141-144.
32 Salmaggi A, Eoli M, Frigerio S. et al. Circulating intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and plasma thrombomodulin levels in glioblastoma patients. Cancer Lett 1999; 146: 169-172.
33 Biguzzi E, Franchi F, Bucciarelli P. et al. Endothelial protein C receptor plasma levels increase in chronic liver disease, while thrombomodulin plasma levels increase only in hepatocellular carcinoma. Thromb Res 2007; 120: 289-293.
34 Goessens BM, van der Graaf Y, Olijhoek JK. et al. The course of vascular risk factors and the occurrence of vascular events in patients with symptomatic peripheral arterial disease. J Vasc Surg 2007; 45: 47-54.
35 Tschudi MR, Barton M, Bersinger NA. et al. Effect of age on kinetics of nitric oxide release in rat aorta and pulmonary artery. J Clin Invest 1996; 98: 899-905.
36 Brown KA, Didion SP, Andresen JJ. et al. Effect of aging, MnSOD deficiency, and genetic background on endothelial function. Evidence for MnSOD haploin-sufficiency. Arterioscler Thromb Vasc Biol 2007; 27: 1941-1946.
37 Schrage WG, Eisenach JH, Joyner MJ. Ageing reduces nitric-oxide- and prostaglandin-mediated vasodilatation in exercising humans. J Physiol 2007; 579: 227-236.