Summary
Measurement of tumor angiogenesis to predict and/or to assess the efficacy of antiangiogenic therapies is mainly based on the evaluation of microvessel density (MVD). We developed a novel flow cytometry procedure to measure circulating endothelial cells (CECs) and circulating endothelial cells progenitors (CECPs) in either preclinical and clinical studies.
Preclinical studies were performed on an animal model of human lymphoma. A trend toward higher CECs values was observed on day 7 and 14 after transplant, and differences vs controls were highly significant on day 21 (p=0.0061). A strong correlation was found between CECs and tumor volumen mice given cyclophosphamide, most of circulating apoptotic cells were hem (r=0.942, p=0.004) and between CECs and tumor-generated VEGF (r=0.669, p=0.02). I atopoietic and not endothelial. Conversely, in mice given endostatin, all of the increase in apoptotic cells was in the endothelial cell compartment.
In a parallel study, we looked for CECs in the peripheral blood of 20 healthy controls and 76 newly diagnosed cancer patients by means of four-color flow cytometry. In breast cancer (n=46) and lymphoma (n=30) patients, both resting and activated CECs were increased by 5 fold (P<O.OOO8 vs control). CECs significantly correlated with plasma levels of VCAM-I and VEGF. Resting and activated CECs were similar to healthy controls in 7 lymphoma patients achieving complete remission after chemotherapy, and activated CECs were found to decrease in 13 breast cancer patients evaluated before and 24h after quadrantectomy.
In conclusion, our findings indicate a close relation between CEC increase and tumor progression, and support CECs evaluation as a clinically relevant, non invasive angiogenesis marker. Furthermore, this assay offers insight into anti-angiogenic activity of different drugs.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
J. Folkman, Angiogenesis in cancer, vascular. rheumatoid and otherdiseases,Nar.Med.1:27–31, 1995.
J. Folkman, P. Hahnfeldt, and J. Hlatky. Cancer: looking outside the genomeNat. Rev. Mol Cell Biol.1: 76–79,2000.
L. Hlatky, P. Hahnfeldt and J. Folkman, Clinical Application of Antiangiogenic Therapy: Microvessel Density, What it does and doesn’t tell us.J Natl Cancer Inst.94:883–893, 2002.
N. Weidner, Angiogenesis as a predictor of clinical outcome in cancer patients.Hum. Pathol.31:403–405, 2000.
A. Solovey, Y. Lin, P. Browne. S. Choong, E. Wayner and Hebbel RP, Circulating endothelial cells in sickle cell anemiaNeu . Engl J Med.337:1584–1590, 1997.
A. Grefte, M. Van der Giessen and W. van Son, The TH. Circulating cytomegalovirus-infected endothelial cells in patients with an active CMV infection.J Infect Dis.167:70–277, 1993.
M. Drancourt, F. George, P. Brouqui. J. Sampol and D. Raoult. Diagnosis of Mediterranean spotted fever by indirect immunofuorescence of Rickettsia conorii in circulating endothelial cells isolated with monoclonal antibody-coated immunomagnetic beadsJ Infect Dis.166:660–663. 1992.
M. Mutin, L Canavy. A. Blann. M. Bory. J. Sampol and F. Dignar-George. Direct evidence of endothelial injury in acute myocardial infarction and unstable angina by demonstration of circulating endothelial cellsBlood.93:2951–2958, 1999.
Y. Lin, D.J. Weisdorf, A. Solovey and R.P. Hebbel, Origins of circulating endothelial cells and endothelial outgrowth from blood.J Clin Invest.105:71–77. 2000.
F. George, P. Poncelet, J.C. Laurent, O. Massot. D. Amoux, N. Lequeux, P. Ambrosi, C. Chicheportiche and J. Sampol, Cytofluorometric detection of human endothelial cells in whole blood using S-Endo I monoclonal antibodyJ Immunol Methods.139:65–75. 1991.
L. Fusetti, A. Gobbi, C. Rabascio. G. Pruneri, N. Carboni. F. Peccatori, G. Martinelli and F. Benolini, Human Myeloid and Lymphoid Malignancies in the Non-Obese Diabetic/Severe Combined Immunodeficiency Mouse Model: Frequency of Apoptotic Cells in Solid Tumors and Efficiency and Speed of Engraftment Correlate with Vascular Endothelial Growth Factor ProductionCancer Res.60:2527–2534, 2000.
F. Bertolini, L. Fusetti, P. Mancuso. A. Gobbi, C. Corsini, P.F. Ferrucci, G. Martinelli and G.Pruneri, Endostatin, an antiangiogenic drug. induces tumor stabilization after chemotherapy or anti-CD20 therapy in a NOD/SCID mouse model of human high-grade non-Hodgkin’s lymphoma.Blood.96:282–287, 2000.
T. Rohem, J. Folkman, T. Browder and M.S. OReilly, Antiangiogenic therapy of experimental cancer does not induce acquired drug resistanceNature.390:404–407, 1997.
N.J. Philpott, A.J.C. Turner, J. Scopes, M. Westby, J.C.W. Marsch, E.C. Gordon-Smith, A. G. Dalgleish and F.M. Gibson, The use of 7-amino actinomycin D in identifying apoptosis: Semplicity of use and broad spectrum of application compared with other techniquesBlood.87:2244–2251, 1996.
J. Folkman,. Incipient angiogenesisJ Natl Cancer Inst.92:94–95, 2000.
D. Hanahan and R.A. Weinberg, The hallmarks of cancerCell.100:57–70, 2000.
B. St Croix, C. Rago, V. Velculescu, G. Traverser. K.E. Romans. E. Montgomery. A. Lal. G.J. Riggins, C. Lengauer, B. Vogelstein and K.W. Kinzler,. Genes expressed in human tumor endotheliumScience.289:1197–1202.2000.
O. Fornas. J. Garcia and J. Petriz, Flow cytometry counting of CD34+ cells in whole bloodNat Med.6:833–836, 2000.
S. Rafii, Circulating endothelial precursors: mystery, reality, and promise. JClin Invest.105:17–19, 2000.
C. Li. I.N. Hampson, L. Hampson, P. Kumar, C. Bemabeu and S. Kumar, CD105 antagonizes the inhibitory signaling of transforming growth factor beta1 on human vascular endothelial cellsFASEB J.14:55–64,2000.
.L. Osborn, C. Hession, R. R.Tizard, C. Vassallo. S. Luhowskyj, G. chi-Rosso and R. Lobb. Direct expression cloning of vascular cell adhesion molecule 1, a cytokine-induced endothelial protein that binds to lymphocytes, Cell59:1203-1211. 1989.
G. Pruneri, F. Bertolini, D. Soligo, N. Carboni. A. Cortelezzi. P.F. Ferrucci. R. Buffa, G. LambertenghiDelilers and F. Pezzella et al. Angiogenesis in myelodysplastic syndromesBr J Cance81:1398–1401.1999
F. Bertolini, M. Paolucci, F. Peccatori, S. Cinieri, A. Agazzi. P.F. Ferrucci. E. Cocorocchio, A. Goldirsch and G. Martinelli, Angiogenic growth factors and endostatin in non-Hodgkin’s lymphomaBr J Haematol106 504–509, 1999.
W. Wynendaele, R. Derua, M.F. Hoylaerts, A. Pawinski, E. Waelkens, E.A. Bmijn, R. Paridaens, W. Merlevede and A.T. van Oosterom, Vascular endothelial growth factor measured in platelet poor plasma allows optimal separation between cancer patients and volunteers: A key to study an angiogenic markerin vivo Ann Oncol 10 965–971, 1999.
T. Asahara, T. Takahashi, H. Masada, C. Kalka, D. Chen, H. Iwaguro. Y. Inai. M. Silver and J.M. lsner, VEGF contributes to postnatal neovascularization by mobilizing bone marrow-derived endothelial progenitor cells.EMBO J. 18 3964–72.1999.
N. Ferrara, and T. Davis Smyth, The biology of vascular endothelial growth factor.Endocrine Rev 18: 425, 1997
Y.S. Chang, E. di Tomaso, D.M. McDonald, R. Jones, R.K. Jain, and L.L. Munn, Mosaic blood vessels in tumors: frequency of cancer cells in contact with flowing bloodProc. Nail. Acad. Sci. USA. 97:1460814613,2000.
C. Li, B. Guo, P.B. Wilson. A. Stewart, G. Byrne, N. Bundred and S. Kumar, Plasma levels of soluble CD105 correlate with metastasis in patients with breast cancerIntl Cancer 89:122–6.2000.
G.J. Byrne. A. Ghellal. J. Iddon, A.D. Blann, V. venizelos, S. Kumar, A. Howell and N.J. Bundred, Serum soluble vascularcelladhesion molecule-I: role as a surrogate marker of angiogenesis.J Nail Cancer Inst 92:1329–36 2000.
D.M. McDonald and A.J.E. Foss, Endothelial cells of tumor vessels: Abnormal but not absentCancer and metastasis reviews J9:109–120. 2000.
J. Folkman, Can mosaic tumor vessels facilitate molecular diagnosis of cancer?Proc. Nail. Acad. Sci. USA 98: 398–400, 2001.
R. Auerbach, N. Akhtar, R.L. Lewis and B. L. Shinners. Angiogenesis assays: problems and pitfallsCancer and metastasis reviews 19: 167–172. 2000.
P. Salven, A. Orpana, L. Teerenhovi. And H. Joensuu, Simultaneous elevation in the serum concentrations of the angiogenic growth factors VEGF and bFGF is an independent predictor of poor prognosis in non-Hodgkin lymphoma: a single-institution study of 200 patientsBlood.96:3712–3718, 2000.
S. Monestiroli, P. Mancuso. A. Burlini. G. Pruneri, C. Dell’Agnola, A. Gobbi, G. Martinelli and F. Bertolini, Kinetics and viability of circulating endothelial cells as surrogate angiogenesis marker in an animal model of human lymphoma.Cancer Res.61 4341–4 2001
P. Mancuso, A. Burlini, G. Pruneri, A. Goldhirsch. G. Martinelli and F. Bertolini, Resting and activated endothelial cells are increased in the peripheral blood of cancer patientsBlood.97:3658–3661, 2001.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2003 Springer Science+Business Media New York
About this chapter
Cite this chapter
Mancuso, P. et al. (2003). Circulating Endothelial Cells as a Novel Marker of Angiogenesis. In: Moldovan, N.I. (eds) Novel Angiogenic Mechanisms. Advances in Experimental Medicine and Biology, vol 522. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-0169-5_9
Download citation
DOI: https://doi.org/10.1007/978-1-4615-0169-5_9
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4613-4951-8
Online ISBN: 978-1-4615-0169-5
eBook Packages: Springer Book Archive