Serum miRNA-based distinct clusters define three groups of breast cancer patients with different clinicopathological and immune characteristics
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Breast cancer (BCa) is a heterogeneous disease with different histological, prognostic and clinical aspects. Therefore, the need for identification of novel biomarkers for diagnosis, prognosis and monitoring of disease, as well as treatment outcome prediction remains at the forefront of research. The search for circulating elements, obtainable by simple peripheral blood withdrawal, which may serve as possible biomarkers, constitutes still a challenge. In the present study, we have evaluated the expression of 6 circulating miRNAs, (miR-16, miR-21, miR-23α, miR-146α, miR-155 and miR-181α), in operable BCa patients, with non-metastatic, invasive ductal carcinoma, not receiving neoadjuvant chemotherapy. These miRNAs, known to be involved in both tumor cell progression and immune pathways regulation, were analyzed in relation to circulating cytokines, tumor immune-cell infiltration and established prognostic clinicopathological characteristics. We have identified three different clusters, with overall low (C1), moderate (C2) or high (C3) expression levels of these six circulating miRNAs, which define three distinct groups of non-metastatic BCa patients characterized by different clinicopathological and immune-related characteristics, with possibly different clinical outcomes. Our data provide the proof-of-principle to support the notion that, up- or down-regulation of the same circulating miRNA may reflect different prognosis in BCa. Nonetheless, the prognostic and/or predictive potential of these three “signatures” needs to be further evaluated in larger cohorts of BCa patients with an, at least, 5-year clinical follow-up.
KeywordsMiRNAs signatures Breast cancer Tumor infiltration Biomarkers Cytokines/chemokines
American Joint Committee on Cancer
Favorable combined immune signatures
Human epidermal growth factor receptor 2
Human leukocyte antigen
Interleukin 1 receptor antagonist
Long noncoding RNA
Major histocompatibility complex
Nuclear factor kappa beta
Receiver operating characteristic
Transforming growth factor beta
T regulatory (cell)
Unfavorable combined immune signatures
Sotirios P. Fortis designed and performed research, analyzed data, and wrote the manuscript; Christoforos K. Vaxevanis performed research, analyzed data and wrote the manuscript; Louisa G. Mahaira, Michael Sofopoulos, Nectaria N. Sotiriadou, Amalia Dinou, Niki Arnogiannaki, Catherine Stavropoulos-Giokas, contributed to experimental design, and performed research; Dimitris Thanos contributed to experimental design and data analysis; Constantin N. Baxevanis contributed to experimental design, data analysis, and wrote the manuscript; Sonia A. Perez supervised the study, contributed to experimental design, data analysis, and wrote the manuscript. All authors read and approved the manuscript.
This study was supported by grant GER_1968 (acronym ISPEBREAST) to Constantin Baxevanis from a bilateral research and innovation cooperation, funded by the General Secretariat for Research and Technology (GSRT) of the Ministry of Education, Research and Religious Affairs of the Hellenic Republic and the German Federal Ministry for Education and Research (BMBF), and a donation to Sonia Perez from the Haegeman-Goossens family, Netherlands.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
The study was approved by the Institutional Review Board of St. Savas Cancer Hospital (IRB-ID 6079/448/10-6-13).
All prospectively enrolled patients signed a written informed consent, approved by the Review Board at St. Savas Cancer Hospital. All data of other retrospectively analyzed patients were obtained from an anonymized database constructed for the purposes of a previous study . Healthy volunteers presented as blood donors at the Blood Collection and Transfusion Department of Saint Savas Hospital. They fulfilled all requirements for blood donation. Volunteers consented verbally, and no personal information was recorded. All procedures were in accordance with the ethical standards of the institutional research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.
- 3.Bai JP, Bell R, Buckman S, Burckart GJ, Eichler HG, Fang KC, Goodsaid FM, Jusko WJ, Lesko LL, Meibohm B, Patterson SD, Puig O, Smerage JB, Snider BJ, Wagner JA, Wang J, Walton MK, Weiner R (2011) Translational biomarkers: from preclinical to clinical a report of 2009 AAPS/ACCP Biomarker Workshop. AAPS J 13(2):274–283. https://doi.org/10.1208/s12248-011-9265-x CrossRefGoogle Scholar
- 4.Ravelli A, Reuben JM, Lanza F, Anfossi S, Cappelletti MR, Zanotti L, Gobbi A, Senti C, Brambilla P, Milani M, Spada D, Pedrazzoli P, Martino M, Bottini A, Generali D,, Marrow Transplantation S (2015) Solid tumor working party of European B. Breast cancer circulating biomarkers: advantages, drawbacks, and new insights. Tumour Biol J Int Soc Oncodev Biol Med 36(9):6653–6665. https://doi.org/10.1007/s13277-015-3944-7 CrossRefGoogle Scholar
- 8.Bahrami A, Aledavood A, Anvari K, Hassanian SM, Maftouh M, Yaghobzade A, Salarzaee O, ShahidSales S, Avan A (2018) The prognostic and therapeutic application of microRNAs in breast cancer: tissue and circulating microRNAs. J Cell Physiol 233(2):774–786. https://doi.org/10.1002/jcp.25813 CrossRefGoogle Scholar
- 12.van Schooneveld E, Wildiers H, Vergote I, Vermeulen PB, Dirix LY, Van Laere SJ (2015) Dysregulation of microRNAs in breast cancer and their potential role as prognostic and predictive biomarkers in patient management. Breast Cancer Res: BCR 17:21. https://doi.org/10.1186/s13058-015-0526-y CrossRefGoogle Scholar
- 18.Eichmuller SB, Osen W, Mandelboim O, Seliger B (2017) Immune modulatory microRNAs involved in tumor attack and tumor immune escape. J Natl Cancer Inst 109 (10). https://doi.org/10.1093/jnci/djx034
- 21.Chen X, Ba Y, Ma L, Cai X, Yin Y, Wang K, Guo J, Zhang Y, Chen J, Guo X, Li Q, Li X, Wang W, Zhang Y, Wang J, Jiang X, Xiang Y, Xu C, Zheng P, Zhang J, Li R, Zhang H, Shang X, Gong T, Ning G, Wang J, Zen K, Zhang J, Zhang CY (2008) Characterization of microRNAs in serum: a novel class of biomarkers for diagnosis of cancer and other diseases. Cell research 18(10):997–1006. https://doi.org/10.1038/cr.2008.282 CrossRefGoogle Scholar
- 24.Fortis SP, Sofopoulos M, Sotiriadou NN, Haritos C, Vaxevanis CK, Anastasopoulou EA, Janssen N, Arnogiannaki N, Ardavanis A, Pawelec G, Perez SA, Baxevanis CN (2017) Differential intratumoral distributions of CD8 and CD163 immune cells as prognostic biomarkers in breast cancer. J Immunother Cancer 5:39. https://doi.org/10.1186/s40425-017-0240-7 CrossRefGoogle Scholar
- 25.Janssen N, Fortis SP, Speigl L, Haritos C, Sotiriadou NN, Sofopoulos M, Arnogiannaki N, Stavropoulos-Giokas C, Dinou A, Perez S, Pawelec G, Baxevanis CN, Shipp C (2017) Peripheral T cell responses to tumour antigens are associated with molecular, immunogenetic and cellular features of breast cancer patients. Breast Cancer Res Treat 161(1):51–62. https://doi.org/10.1007/s10549-016-4037-z CrossRefGoogle Scholar
- 27.Anastasopoulou EA, Voutsas IF, Keramitsoglou T, Gouttefangeas C, Kalbacher H, Thanos A, Papamichail M, Perez SA, Baxevanis CN (2015) A pilot study in prostate cancer patients treated with the AE37 Ii-key-HER-2/neu polypeptide vaccine suggests that HLA-A*24 and HLA-DRB1*11 alleles may be prognostic and predictive biomarkers for clinical benefit. Cancer Immunol Immunother 64(9):1123–1136. https://doi.org/10.1007/s00262-015-1717-1 CrossRefGoogle Scholar
- 28.Stokidis S, Dinou A, Fortis SP, Vaxevanis CK, Konstantellou M, Stavropoulos-Giokas C, Perez SA, Baxevanis CN (2017) The impact of HLA-A*02 and HLA-A*24 allele expression in prostate cancer prognosis In: 3rd Symposium on Advances in Cancer Immunology and Immunotherapy, Athens, Greece, November 2–4. https://doi.org/10.13140/RG.2.2.34617.47202
- 29.Vaxevanis C, Anastasopoulou E, Tzonis P, Ardavanis A, Baxevanis CN, Peoples GE, Perez SA (2017) An IFN-γ response-based algorithm with predictive potential in AE37-vaccinated breast cancer patients In: 3rd Symposium on Advances in Cancer Immunology and Immunotherapy, Athens, Greece, November 2–4 2017. https://doi.org/10.13140/RG.2.2.24551.14248
- 31.Jurkovicova D, Smolkova B, Magyerkova M, Sestakova Z, Kajabova VH, Kulcsar L, Zmetakova I, Kalinkova L, Krivulcik T, Karaba M, Benca J, Sedlackova T, Minarik G, Cierna Z, Danihel L, Mego M, Chovanec M, Fridrichova I (2017) Down-regulation of traditional oncomiRs in plasma of breast cancer patients. Oncotarget 8(44):77369–77384. https://doi.org/10.18632/oncotarget.20484 CrossRefGoogle Scholar
- 33.Perez EA, Romond EH, Suman VJ, Jeong JH, Sledge G, Geyer CE Jr, Martino S, Rastogi P, Gralow J, Swain SM, Winer EP, Colon-Otero G, Davidson NE, Mamounas E, Zujewski JA, Wolmark N (2014) Trastuzumab plus adjuvant chemotherapy for human epidermal growth factor receptor 2-positive breast cancer: planned joint analysis of overall survival from NSABP B-31 and NCCTG N9831. J Clin Oncol 32(33):3744–3752. https://doi.org/10.1200/JCO.2014.55.5730 CrossRefGoogle Scholar
- 39.Lu LF, Thai TH, Calado DP, Chaudhry A, Kubo M, Tanaka K, Loeb GB, Lee H, Yoshimura A, Rajewsky K, Rudensky AY (2009) Foxp3-dependent microRNA155 confers competitive fitness to regulatory T cells by targeting SOCS1 protein. Immunity 30(1):80–91. https://doi.org/10.1016/j.immuni.2008.11.010 CrossRefGoogle Scholar
- 41.Perry MM, Williams AE, Tsitsiou E, Larner-Svensson HM, Lindsay MA (2009) Divergent intracellular pathways regulate interleukin-1beta-induced miR-146a and miR-146b expression and chemokine release in human alveolar epithelial cells. FEBS Lett 583(20):3349–3355. https://doi.org/10.1016/j.febslet.2009.09.038 CrossRefGoogle Scholar
- 44.Nagata Y, Hanagiri T, Mizukami M, Kuroda K, Shigematsu Y, Baba T, Ichiki Y, Yasuda M, So T, Takenoyama M, Sugio K, Nagashima A, Yasumoto K (2009) Clinical significance of HLA class I alleles on postoperative prognosis of lung cancer patients in Japan. Lung Cancer 65(1):91–97. https://doi.org/10.1016/j.lungcan.2008.10.012 CrossRefGoogle Scholar