Breast Cancer Research and Treatment

, Volume 169, Issue 1, pp 83–92 | Cite as

Circulating small-sized endothelial microparticles as predictors of clinical outcome after chemotherapy for breast cancer: an exploratory analysis

  • Elisa García Garre
  • Ginés Luengo Gil
  • Silvia Montoro García
  • Enrique Gonzalez Billalabeitia
  • Marta Zafra Poves
  • Elena García Martinez
  • Vanessa Roldán Schilling
  • Esther Navarro Manzano
  • Alejandra Ivars Rubio
  • Gregory Y. H. Lip
  • Francisco Ayala de la Peña
Clinical trial



Therapeutic exploitation of angiogenesis in breast cancer has been limited by the lack of reliable biomarkers. Circulating small-sized endothelial microparticles (sEMP) are likely to play a significant role as messengers of angiogenesis. Higher levels of EMP have been observed in cancer patients, but their prognostic value in breast cancer is unknown. Our aim was to determine the value of circulating sEMP as a marker of response to chemotherapy in breast cancer.


We included patients with breast cancer treated with neoadjuvant or first-line chemotherapy. Baseline and post-treatment circulating sEMP (CD144+) were quantified using a flow cytometer approach specifically designed for analysis of small-sized particles (0.1–0.5 μm). Small-sized EMP response was defined as a post-treatment decrease of sEMP larger than the median decrease of sEMP after chemotherapy. Baseline and post-chemotherapy VEGFA levels were determined with ELISA.


Forty-four breast cancer patients were included (19 with metastatic and 25 with locally advanced disease). Median levels of sEMP decreased after chemotherapy (P = 0.005). Response to chemotherapy showed a non-significant trend to associate with sEMP response (P = 0.056). A sEMP response was observed in 51% of patients and was associated with better overall survival (HR 0.18; 95% CI 0.04–0.87; P = 0.02) and progression free survival (HR 0.30; 95% CI 0.09–0.99; P = 0.04) in the group of women with metastatic disease. Post-chemotherapy decrease of VEGFA levels was not associated with breast cancer prognosis.


Our results did not support sEMP as a marker of response to chemotherapy. However, our exploratory analysis suggests that in patients with metastatic breast cancer, the decrease of sEMP levels after chemotherapy is associated with better overall and disease free survival and might be superior to VEGFA levels as an angiogenesis-related prognostic marker.


Small-sized microparticles Endothelial Breast cancer Chemotherapy VEGFA 



Breast cancer




Hormonal receptors


Locally advanced breast cancer


Metastatic breast cancer




Extracellular microvesicles


Small-sized endothelial microparticles


Triple negative breast cancer


Vascular and endothelial growth factor A



We acknowledge technical assistance by José A. López Oliva (Clinical Research Unit, Department of Hematology and Medical Oncology, Hospital Universitario Morales Meseguer) and Alberto Martínez (Biobancmur, Nodo 3, Hospital Universitario Morales Meseguer, IMIB Arrixaca). This work was partially funded by “Calasparra se mueve,” a research-funding initiative inspired by women from Calasparra (Murcia), Spain. SMG was supported by the People Program (Marie Curie Actions) of the European Union’s Seventh Framework Program (FP7/2007-2013) under REA Grant Agreement No. 608765.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in this study were in accordance with the ethical standards of the institution and with the 1964 Helsinki declaration and its later amendments.

Informed consent

Informed consent was obtained from all individual participants included in the study and the Institutional Review Board of Hospital Morales Meseguer approved the study.

Supplementary material

10549_2017_4656_MOESM1_ESM.pdf (754 kb)
Electronic supplementary material 1 (PDF 754 kb)


  1. 1.
    Miles DW, Diéras V, Cortés J et al (2013) First-line bevacizumab in combination with chemotherapy for HER2-negative metastatic breast cancer: pooled and subgroup analyses of data from 2447 patients. Ann Oncol 24:2773–2780CrossRefPubMedGoogle Scholar
  2. 2.
    Ghosh S, Sullivan CAW, Zerkowski MP et al (2008) High levels of vascular endothelial growth factor and its receptors (VEGFR-1, VEGFR-2, neuropilin-1) are associated with worse outcome in breast cancer. Hum Pathol 39:1835–1843CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Berns EMJJ, Klijn JGM, Look MP et al (2003) Combined vascular endothelial growth factor and TP53 status predicts poor response to tamoxifen therapy in estrogen receptor-positive advanced breast cancer. Clin Cancer Res 9:1253–1258PubMedGoogle Scholar
  4. 4.
    Foekens JA, Peters HA, Grebenchtchikov N et al (2001) High tumor levels of vascular endothelial growth factor predict poor response to systemic therapy in advanced breast cancer. Cancer Res 61:5407–5414PubMedGoogle Scholar
  5. 5.
    Lissoni P, Rovelli F, Malugani F et al (2003) Changes in circulating VEGF levels in relation to clinical response during chemotherapy for metastatic cancer. Int J Biol Markers 18:152–155CrossRefPubMedGoogle Scholar
  6. 6.
    dos Santos LV, Cruz MR, de Lopes G, Lima JP (2015) VEGF-A levels in bevacizumab-treated breast cancer patients: a systematic review and meta-analysis. Breast Cancer Res Treat 151:481–489CrossRefPubMedGoogle Scholar
  7. 7.
    Al-Nedawi K, Rak J, D’Asti E et al (2011) Microvesicles as mediators of intercellular communication in cancer—the emerging science of cellular “debris”. Semin Immunopathol 33:455–467CrossRefPubMedGoogle Scholar
  8. 8.
    Lozito TP, Tuan RS (2014) Endothelial and cancer cells interact with mesenchymal stem cells via both microparticles and secreted factors. J Cell Mol Med 18:2372–2384CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Piccin A, Murphy WG, Smith OP (2007) Circulating microparticles: pathophysiology and clinical implications. Blood Rev 21:157–171CrossRefPubMedGoogle Scholar
  10. 10.
    Orozco AF, Lewis DE (2010) Flow cytometric analysis of circulating microparticles in plasma. Cytometry A 77:502–514CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Melo SA, Luecke LB, Kahlert C et al (2015) Glypican-1 identifies cancer exosomes and detects early pancreatic cancer. Nature 523:177–182CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Valenti R, Huber V, Iero M et al (2007) Tumor-released microvesicles as vehicles of immunosuppression. Cancer Res 67:2912–2915CrossRefPubMedGoogle Scholar
  13. 13.
    Martinez MC, Andriantsitohaina R (2011) Microparticles in Angiogenesis: therapeutic Potential. Circ Res 109:110–119CrossRefPubMedGoogle Scholar
  14. 14.
    Ribeiro MF, Zhu H, Millard RW, Fan G-C (2013) Exosomes function in pro- and anti-angiogenesis. Curr Angiogenes 2:54–59PubMedPubMedCentralGoogle Scholar
  15. 15.
    Taraboletti G, D’Ascenzo S, Borsotti P et al (2002) Shedding of the matrix metalloproteinases MMP-2, MMP-9, and MT1-MMP as membrane vesicle-associated components by endothelial cells. Am J Pathol 160:673–680CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Janowska-Wieczorek A, Marquez-Curtis LA, Wysoczynski M, Ratajczak MZ (2006) Enhancing effect of platelet-derived microvesicles on the invasive potential of breast cancer cells. Transfusion 46:1199–1209CrossRefPubMedGoogle Scholar
  17. 17.
    Millimaggi D, Mari M, D’Ascenzo S et al (2007) Tumor vesicle-associated CD147 modulates the angiogenic capability of endothelial cells. Neoplasia 9:349–357CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Hoyer FF, Nickenig G, Werner N (2010) Microparticles–messengers of biological information. J Cell Mol Med 14:2250–2256CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Sheremata WA, Jy W, Delgado S et al (2006) Interferon-beta1a reduces plasma CD31 + endothelial microparticles (CD31 + EMP) in multiple sclerosis. J Neuroinflammation 3:23CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Montoro-García S, Shantsila E, Tapp LD et al (2013) Small-size circulating microparticles in acute coronary syndromes: relevance to fibrinolytic status, reparative markers and outcomes. Atherosclerosis 227:313–322CrossRefPubMedGoogle Scholar
  21. 21.
    Tseng C-C, Wang C-C, Chang H-C et al (2013) Levels of circulating microparticles in lung cancer patients and possible prognostic value. Dis Markers 35:301–310CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Laresche C, Pelletier F, Garnache-Ottou F et al (2013) Increased levels of circulating microparticles are associated with increased procoagulant activity in patients with cutaneous malignant melanoma. J Invest Dermatol 134:176–182CrossRefPubMedGoogle Scholar
  23. 23.
    Wang C-C, Tseng C-C, Hsiao C-C et al (2014) Circulating endothelial-derived activated microparticle: a useful biomarker for predicting 1-year mortality in patients with advanced non-small cell lung cancer. Biomed Res Int 2014:173401PubMedPubMedCentralGoogle Scholar
  24. 24.
    Campello E, Spiezia L, Radu CM et al (2011) Endothelial, platelet, and tissue factor-bearing microparticles in cancer patients with and without venous thromboembolism. Thromb Res 127:473–477CrossRefPubMedGoogle Scholar
  25. 25.
    Toth B, Nieuwland R, Liebhardt S et al (2008) Circulating microparticles in breast cancer patients: a comparative analysis with established biomarkers. Anticancer Res 28:1107–1112PubMedGoogle Scholar
  26. 26.
    Eisenhauer EA, Therasse P, Bogaerts J et al (2009) New response evaluation criteria in solid tumours: revised RECIST guideline (version 1.1). Eur J Cancer 45:228–247CrossRefPubMedGoogle Scholar
  27. 27.
    McShane LM, Altman DG, Sauerbrei W et al (2006) REporting recommendations for tumor MARKer prognostic studies (REMARK). Breast Cancer Res Treat 100:229–235CrossRefPubMedGoogle Scholar
  28. 28.
    Montoro-García S, Shantsila E, Orenes-Piñero E et al (2012) An innovative flow cytometric approach for small-size platelet microparticles: influence of calcium. Thromb Haemost 108:373–383CrossRefPubMedGoogle Scholar
  29. 29.
    Aharon A, Sabbah A, Ben-shaul S et al (2017) Chemotherapy administration to breast cancer patients affects extracellular vesicles thrombogenicity and function. Oncotarget 8:63265–63280CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Shantsila E, Montoro-García S, Gallego P, Lip GYH (2014) Circulating microparticles: challenges and perspectives of flow cytometric assessment. Thromb Haemost 111:1009–1014CrossRefPubMedGoogle Scholar
  31. 31.
    Sabatier F, Camoin-Jau L, Anfosso F et al (2009) Circulating endothelial cells, microparticles and progenitors: key players towards the definition of vascular competence. J Cell Mol Med 13:454–471CrossRefPubMedGoogle Scholar
  32. 32.
    Lechner D, Kollars M, Gleiss A et al (2007) Chemotherapy-induced thrombin generation via procoagulant endothelial microparticles is independent of tissue factor activity. J Thromb Haemost 5:2445–2452CrossRefPubMedGoogle Scholar
  33. 33.
    Bovy N, Blomme B, Frères P et al (2015) Endothelial exosomes contribute to the antitumor response during breast cancer neoadjuvant chemotherapy via microRNA transfer. Oncotarget 6:10253–10266CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Andre N, Cointe S, Barlogis V, Arnaud L et al (2015) Maintenance chemotherapy in children with aLL exerts metronomic-like thrombospondin-1 associated anti-endothelial effect. Oncotarget 6:23008–23014CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Tang J-H, Zhao J-H, Lu J-W et al (2011) Circulating levels of angiogenic cytokines in advanced breast cancer patients with system chemotherapy and their potential value in monitoring disease course. J Cancer Res Clin Oncol 137:55–63CrossRefPubMedGoogle Scholar
  36. 36.
    Munster M, Fremder E, Miller V et al (2014) Anti-VEGF-A affects the angiogenic properties of tumor-derived microparticles. PLoS ONE 9:e95983. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Elisa García Garre
    • 1
    • 2
  • Ginés Luengo Gil
    • 1
    • 2
    • 3
  • Silvia Montoro García
    • 4
    • 5
  • Enrique Gonzalez Billalabeitia
    • 1
    • 2
    • 4
  • Marta Zafra Poves
    • 1
    • 2
  • Elena García Martinez
    • 1
    • 2
    • 4
  • Vanessa Roldán Schilling
    • 1
    • 2
    • 3
  • Esther Navarro Manzano
    • 1
    • 2
    • 3
  • Alejandra Ivars Rubio
    • 1
    • 2
  • Gregory Y. H. Lip
    • 5
  • Francisco Ayala de la Peña
    • 1
    • 2
    • 3
  1. 1.Servicio de Hematología y Oncología MédicaHospital Universitario Morales MeseguerMurciaSpain
  2. 2.Instituto Murciano de Investigación Biosanitaria (IMIB-Arrixaca)MurciaSpain
  3. 3.Universidad de MurciaMurciaSpain
  4. 4.Universidad Católica San Antonio de MurciaMurciaSpain
  5. 5.Institute of Cardiovascular SciencesUniversity of BirminghamBirminghamUK

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