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High expression of CCL2 in tumor cells and abundant infiltration with CD14 positive macrophages predict early relapse in breast cancer

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Abstract

Macrophages are important for the function of the innate immune system, and in solid tumors, they represent a significant proportion of the tumor mass. Tumor-associated macrophages (TAM) have a M2 phenotype and show a multitude of pro-tumoral functions, promoting tumor cell survival, proliferation, and dissemination. CCL2, synthesized by tumor and stromal cells, initiates a chemokine cascade inducing these processes. We studied by immunohistochemistry (IHC) the frequency of TAMs and CCL2 expressing cells in three groups of primary tumor (PT)-recurrence (R) pairs, where relapse was recorded within 2 years (group 1), between 5 and 10 years (group 2), and after 10 years (group 3). In our study all established breast cancers were heavily infiltrated by CD68 positive cells. Both in PTs and in R lesions the infiltration was more abundant in the peritumoral than in the intratumoral stroma. The mean frequency of M2 marker and CD14 positive cells in the intratumoral stroma and CCL2 expressing tumor cells was higher in the Rs as compared to the corresponding PTs. In PTs, a high frequency of CD14 positive cells and a high expression of CCL2 by tumor cells was associated with an early recurrence. The findings support the current understanding of immune cell orchestrated development, progression and metastatic spread of breast cancer. Our study showed that a high frequency of CCL2 positive tumor cells and CD14 positive TAMs are significant risk factors for rapid tumor recurrence. Potential targets for intervention are discussed.

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References

  1. Gouon-Evans V, Rothenbeg ME, Pollard JW (2000) Postnatal mammary gland development requires macrophages and eosinophils. Development 127:2269–2282

    CAS  PubMed  Google Scholar 

  2. Pollard JW (2009) Trophic macrophages in development and disease. Nat Rev Immunol 9:259–270

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Mantovani A, Sozzani S, Locati M, Allavena P, Sica A (2002) Macrophage polarization: tumor-associated macrophages as a paradigm for polarized M2 mononuclear phagocytes. Trends Immunol 23:549–555

    Article  CAS  PubMed  Google Scholar 

  4. Pollard JW (2004) Tumor-educated macrophages promote tumor progression and metastasis. Nat Rev Cancer 4:71–78

    Article  CAS  PubMed  Google Scholar 

  5. Mantovani A, Sica A, Sozzani S, Allavena P, Vecchi A, Locati M (2004) The chemokine system in diverse forms of macrophage activation and polarization. Trends Immunol 25:677–686

    Article  CAS  PubMed  Google Scholar 

  6. Zawada AM, Rogacev KS, Rotter B, Winter P, Marell RR, Fliser D, Heine GH (2011) SuperSAGE evidence for CD14++CD16+ monocytes as a third monocyte subset. Blood 118(12):e50–e61

    Article  CAS  PubMed  Google Scholar 

  7. Mosser DM, Edwards JP (2008) Exploring the full spectrum of macrophage activation. Nat Rev Immunol 8:958–969

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Ambarus CA, Krausz S, van Eijk M, Hamann J, Radstake TRDJ, Reedquist KA, Tak PP, Baeten DLP (2012) Systematic validation of specific phenotypic markers for in vitro polarized human macrophages. J Immunol Methods 375:196–206

    Article  CAS  PubMed  Google Scholar 

  9. Solinas G, Germano G, Mantovani A, Allavena P (2009) Tumor-associated macrophages (TAM) as major players of the cancer-related inflammation. J Leukoc Biol 86(5):1065–1073

    Article  CAS  PubMed  Google Scholar 

  10. Pollard JW (2008) Macrophages define the invasive microenvironment in breast cancer. J Leukoc Biol 84:623–630

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Qian B, Li J, Zhang H et al (2011) CCL2 recruits inflammatory monocytes to facilitate breast-tumour metastasis. Nature 475:222–225

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Movahedi K, Laoui D, Gysemans C, Baeten M, Stange G, van den Bossche J, Mack M, Pipeleers D, in't Veld P, de Baetselier P, van Ginderachter JA (2010) Different tumor microenvironments contain functionally distinct subsets of macrophages derived from Ly6C(high) monocytes. Cancer Res 70:5728–5739

    Article  CAS  PubMed  Google Scholar 

  13. Lu X, Kang Y (2009) Chemokine (C-C motif) ligand 2 engages CCR2 + stromal cells of monocytic origin to promote breast cancer metastasis to lung and bone. J Biol Chem 284:29087–29096

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Li M, Knight DA, Snyder LA, Smyth MJ, Stewart TJ (2013) A role for CCL2 in both tumor progression and immunosurveillance. OncoImmunology 2(7):e25474

    Article  PubMed  PubMed Central  Google Scholar 

  15. Zhu X, Fujita L, Snyder A, Okada H (2011) Systemic delivery of neutralizing antibody targeting CCL2 for glioma therapy. J Neuro-Oncol 104:83–92

    Article  CAS  Google Scholar 

  16. Hussein MR, Hassan HI (2006) Analysis of the mononuclear inflammatory cell infiltrate in the normal breast, benign proliferative breast disease, in situ and infiltrating ductal breast carcinomas: preliminary observations. J Clin Pathol 59:972–977

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Volodko N, Reiner A, Rudas M, Jakesz R (1998) Tumour-associated macrophages in breast cancer and their prognostic correlations. Breast 7:99–105

    Article  Google Scholar 

  18. Medrek C, Ponten F, Jirstrom K, Leandersson K (2012) The presence of tumor associated macrophages in tumor stroma as a prognostic marker for breast cancer patients. BMC Cancer 12:306–315

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Leek RD, Lewis CE, Whitehouse R, Greenall M, Clarke J, Harris AL (1996) Association of macrophage infiltration with angiogenesis and prognosis in invasive breast carcinoma. Cancer Res 56:4625–4629

    CAS  PubMed  Google Scholar 

  20. Joensuu K, Leidenius M, Kero M, Andersson LC, Horwitz KB, Heikkilä P (2013) ER, PR, HER2, Ki67 and CK5 in early and late relapsing breast cancer - reduced CK5 expression in metastases. Breast Cancer: Bacic Clin Res 7:23–34

    Google Scholar 

  21. Laoui D, Movahedi K, van Overmeire E, van den Bossche J, Schouppe E, Mommer C, Nikolaou A, Morias Y, de Baetselier P, van Ginderachter JA (2011) Tumor-associated macrophages in breast cancer: distinct subsets, distinct functions. Int J Dev Biol 55:861–867

    Article  PubMed  Google Scholar 

  22. Yang J, Zhang L, Yu C, Yang X-F, Wang H (2014) Monocyte and macrophage differentiation: circulation inflammatory monocyte as biomarker for inflammatory diseases. Biomarker Research 2:1–9

    Article  PubMed  PubMed Central  Google Scholar 

  23. Loberg RD, Ying C, Craig M, Day LL, Sargent E, Neeley C, Wojno K, Snyder LA, Yan L, Pienta KJ (2007) Targeting CCL2 with systemic delivery of neutralizing antibodies induces prostate cancer tumor regression in vivo. Cancer Res 67:9417–9424

    Article  CAS  PubMed  Google Scholar 

  24. Conti I, Rollins BJ (2004) CCL2 (monocyte chemoattractant protein-1) and cancer. Semin Cancer Biol 14:149–154

    Article  CAS  PubMed  Google Scholar 

  25. Roca H, Varsos ZS, Sud S, Craig MJ, Ying C, Pienta KJ (2009) CCL2 and Interleukin-6 promote survival of human CD11b + peripheral blood mononuclear cells and induce M2-type macrophage polarization. J Biol Chem 284(49):34342–34354

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Querzoli P, Albonico G, Ferretti S, Rnaldi R, Magri E, Indelli M, Nenci I (1996) MIB-1 proliferative activity in invasive breast cancer measured by image analysis. J Clin Pathol 49(11):926–930

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Taneja P, Maglic KF, Zhu S, Kendig RD, Fry EA, Inoue K (2010) Classical and novel prognostic markers for breast cancer and their clinical significance. Clin Med Insights Oncol 4:15–34

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Allavena P, Signorelli M, Chiappa M et al (2005) Anti-inflammatory properties of the novel antitumor agent yondelis (trabectedin): inhibition of macrophage differentiation and cytokine production. Cancer Res 65:2964–2971

    Article  CAS  PubMed  Google Scholar 

  29. Frow EK, Reckless J, Grainger DJ (1999) Tools for anti-inflammatory drug design: in vitro models of leukocyte migration. Med Res Rev 24(3):276–282

    Article  CAS  Google Scholar 

  30. Nywening TM, Wang-Gillam A, Sanford DA et al (2016) Phase 1b study targeting tumour associated macrophages with CCR2 inhibition plus FOLFIRINOX in locally advanced and borderline resectable pancreatic cancer. Lancet Oncol 17(5):651–662. https://doi.org/10.1016/S1470-2045(16)00078-4

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Giraudo E, Inoue M, Hanahan D (2004) An amino-bisphosphonate targets MMP-9-expressing macrophages and angiogenesis to impair cervical carcinogenesis. J Clin Invest 114:623–633

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Kitamura T, Qian B-Z, Soong D, Cassetta L, Noy R, Sugano G, Kato Y, Li J, Pollard JW (2015) CCL2-induced chemokine cascade promotes breast cancer metastasis by enhancing retention of metastasis-associated macrophages. J Exp Med 212(7):1043–1059

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Mathenge E, Dean C, Clements D et al (2014) Core needle biopsy of breast Cancer tumors increases distant metastases in a mouse model. Neoplasia 16:950–960

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We thank Eija Heiliö for her excellent technical assistance, and Antti Nevanlinna, Msc, for facilitating the statistical analysis.

Funding

This work was supported by the Helsinki University Central Hospital Research Foundation, the Finnish Cancer Foundation, the Sigrid Juselius Foundation, and the Finnish Breast Cancer Group.

Author information

Authors and Affiliations

  1. Department of Pathology and HUSLAB, Helsinki University Hospital and University of Helsinki, FIN-00290, Helsinki, Finland

    Marja Heiskala, Kristiina Joensuu & Päivi Heikkilä

  2. Breast Surgery Unit, Comprehensive Cancer Center Helsinki University Hospital, P.O Box 163 , 00290 HUS, Helsinki, Finland University of Helsinki, Central Hospital, FIN-00290, Helsinki, Finland

    Marjut Leidenius

Authors
  1. Marja Heiskala
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  2. Marjut Leidenius
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  3. Kristiina Joensuu
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  4. Päivi Heikkilä
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Contributions

Marja Heiskala: doing the literature search and planning the study, evaluating the samples, writing the article, and preparing the graphics

Marjut Leidenius: planning the study, writing the article

Kristiina Joensuu: planning the study, responsible for the collection and handling of the samples, evaluating the immunohistochemical stainings, making the statistical work, and writing the article

Päivi Heikkilä: planning the study and writing the article, analyzing the stainings.

Corresponding author

Correspondence to Kristiina Joensuu.

Ethics declarations

The Ethics Committee of the Helsinki University Central Hospital approved the study protocol.

Conflict of interest

The authors declare that they have no conflict of interest.

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Heiskala, M., Leidenius, M., Joensuu, K. et al. High expression of CCL2 in tumor cells and abundant infiltration with CD14 positive macrophages predict early relapse in breast cancer. Virchows Arch 474, 3–12 (2019). https://doi.org/10.1007/s00428-018-2461-7

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  • DOI: https://doi.org/10.1007/s00428-018-2461-7

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