Breast Cancer Research and Treatment

, Volume 148, Issue 1, pp 41–59 | Cite as

Infiltrating S100A8+ myeloid cells promote metastatic spread of human breast cancer and predict poor clinical outcome

  • Katherine Drews-ElgerEmail author
  • Elizabeth Iorns
  • Alexandra Dias
  • Philip Miller
  • Toby M. Ward
  • Sonja Dean
  • Jennifer Clarke
  • Adriana Campion-Flora
  • Daniel Nava Rodrigues
  • Jorge S. Reis-Filho
  • James M. Rae
  • Dafydd Thomas
  • Deborah Berry
  • Dorraya El-Ashry
  • Marc E. Lippman
Preclinical Study


The mechanisms by which breast cancer (BrC) can successfully metastasize are complex and not yet fully understood. Our goal was to identify tumor-induced stromal changes that influence metastatic cell behavior, and may serve as better targets for therapy. To identify stromal changes in cancer-bearing tissue, dual-species gene expression analysis was performed for three different metastatic BrC xenograft models. Results were confirmed by immunohistochemistry, flow cytometry, and protein knockdown. These results were validated in human clinical samples at the mRNA and protein level by retrospective analysis of cohorts of human BrC specimens. In pre-clinical models of BrC, systemic recruitment of S100A8+ myeloid cells—including myeloid-derived suppressor cells (MDSCs)—was promoted by tumor-derived factors. Recruitment of S100A8+ myeloid cells was diminished by inhibition of tumor-derived factors or depletion of MDSCs, resulting in fewer metastases and smaller primary tumors. Importantly, these MDSCs retain their ability to suppress T cell proliferation upon co-culture. Secretion of macrophage inhibitory factor (MIF) activated the recruitment of S100A8+ myeloid cells systemically. Inhibition of MIF, or depletion of MDSCs resulted in delayed tumor growth and lower metastatic burden. In human BrC specimens, increased mRNA and protein levels of S100A8+ infiltrating cells are highly associated with poor overall survival and shorter metastasis free survival of BrC patients, respectively. Furthermore, analysis of nine different human gene expression datasets confirms the association of increased levels of S100A8 transcripts with an increased risk of death. Recruitment of S100A8+ myeloid cells to primary tumors and secondary sites in xenograft models of BrC enhances cancer progression independent of their suppressive activity on T cells. In clinical samples, infiltrating S100A8+ cells are associated with poor overall survival. Targeting these molecules or associated pathways in cells of the tumor microenvironment may translate into novel therapeutic interventions and benefit patient outcome.


S100A8 Myeloid-derived suppressor cells (MDSCs) Inflammation and tumor development Cytokines Molecular markers of metastasis and progression 



The authors would like to thank the members of the Oncogenomics Core Facility, Flow Cytometry Core Facility, and the Division of Veterinary Resources at the University of Miami Miller School of Medicine for their assistance during the course of the study. The authors would also like to thank Nanette Bishopric, MD and Barry I. Hudson PhD for helpful discussion of the manuscript. This Project was funded by Breast Cancer Research Foundation (BrCRF) awards to MEL and JMR. Part of these studies was conducted at the Lombardi Comprehensive Cancer Center Histopathology and Tissue Shared Resource which is supported in part by NIH/NCI Grant P30-CA051008. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Cancer Institute or the National Institutes of Health.

Conflict of interest

All authors have no conflicts of interest to declare.

Supplementary material

10549_2014_3122_MOESM1_ESM.pdf (733 kb)
Supplementary material 1 (PDF 733 kb)


  1. 1.
    Hanahan D, Weinberg RA (2011) Hallmarks of cancer: the next generation. Cell 144(5):646–674. doi: 10.1016/j.cell.2011.02.013 PubMedCrossRefGoogle Scholar
  2. 2.
    Finak G, Bertos N, Pepin F, Sadekova S, Souleimanova M, Zhao H, Chen H, Omeroglu G, Meterissian S, Omeroglu A, Hallett M, Park M (2008) Stromal gene expression predicts clinical outcome in breast cancer. Nat Med 14(5):518–527. doi: 10.1038/nm1764 PubMedCrossRefGoogle Scholar
  3. 3.
    Iorns E, Drews-Elger K, Ward TM, Dean S, Clarke J, Berry D, El-Ashry D, Lippman M (2012) A new mouse model for the study of human breast cancer metastasis. PLoS ONE 7(10):e47995. doi: 10.1371/journal.pone.0047995 PubMedCrossRefPubMedCentralGoogle Scholar
  4. 4.
    Sartelet H, Durrieu L, Fontaine F, Nyalendo C, Haddad E (2012) Description of a new xenograft model of metastatic neuroblastoma using NOD/SCID/Il2rg null (NSG) mice. In Vivo 26(1):19–29PubMedGoogle Scholar
  5. 5.
    Quintana H, Piskounova E, Shackleton M, Weinberg D, Eskiocak U, Fullen DR, Johnson TM, Morrison SJ (2012) Human melanoma metastasis in NSG mice correlates with clinical outcome in patients. Sci Transl Med 4(159):159ra149. doi: 10.1126/scitranslmed.3004599 PubMedCrossRefGoogle Scholar
  6. 6.
    Iorns E, Clarke J, Ward T, Dean S, Lippman M (2012) Simultaneous analysis of tumor and stromal gene expression profiles from xenograft models. Breast Cancer Res Treat 131(1):321–324. doi: 10.1007/s10549-011-1784-8 PubMedCrossRefGoogle Scholar
  7. 7.
    Murdoch C, Muthana M, Coffelt SB, Lewis CE (2008) The role of myeloid cells in the promotion of tumour angiogenesis. Nat Rev Cancer 8(8):618–631. doi: 10.1038/nrc2444 PubMedCrossRefGoogle Scholar
  8. 8.
    Nagaraj S, Gabrilovich DI (2010) Myeloid-derived suppressor cells in human cancer. Cancer J 16(4):348–353. doi: 10.1097/PPO.0b013e3181eb3358 PubMedCrossRefGoogle Scholar
  9. 9.
    Youn JI, Nagaraj S, Collazo M, Gabrilovich DI (2008) Subsets of myeloid-derived suppressor cells in tumor-bearing mice. J Immunol 181(8):5791–5802PubMedCrossRefPubMedCentralGoogle Scholar
  10. 10.
    Cuenca AG, Delano MJ, Kelly-Scumpia KM, Moreno C, Scumpia PO, Laface DM, Heyworth PG, Efron PA, Moldawer LL (2011) A paradoxical role for myeloid-derived suppressor cells in sepsis and trauma. Mol Med 17(3–4):281–292. doi: 10.2119/molmed.2010.00178 PubMedPubMedCentralGoogle Scholar
  11. 11.
    Diaz-Montero CM, Salem ML, Nishimura MI, Garrett-Mayer E, Cole DJ, Montero AJ (2009) Increased circulating myeloid-derived suppressor cells correlate with clinical cancer stage, metastatic tumor burden, and doxorubicin-cyclophosphamide chemotherapy. Cancer Immunol Immunother 58(1):49–59. doi: 10.1007/s00262-008-0523-4 PubMedCrossRefPubMedCentralGoogle Scholar
  12. 12.
    Srivastava MK, Sinha P, Clements VK, Rodriguez P, Ostrand-Rosenberg S (2010) Myeloid-derived suppressor cells inhibit T-cell activation by depleting cystine and cysteine. Cancer Res 70(1):68–77. doi: 10.1158/0008-5472.CAN-09-2587 PubMedCrossRefPubMedCentralGoogle Scholar
  13. 13.
    Bunt SK, Yang L, Sinha P, Clements VK, Leips J, Ostrand-Rosenberg S (2007) Reduced inflammation in the tumor microenvironment delays the accumulation of myeloid-derived suppressor cells and limits tumor progression. Cancer Res 67(20):10019–10026. doi: 10.1158/0008-5472.CAN-07-2354 PubMedCrossRefGoogle Scholar
  14. 14.
    Sinha P, Clements VK, Bunt SK, Albelda SM, Ostrand-Rosenberg S (2007) Cross-talk between myeloid-derived suppressor cells and macrophages subverts tumor immunity toward a type 2 response. J Immunol 179(2):977–983PubMedCrossRefGoogle Scholar
  15. 15.
    Ostrand-Rosenberg S, Sinha P (2009) Myeloid-derived suppressor cells: linking inflammation and cancer. J Immunol 182(8):4499–4506. doi: 10.4049/jimmunol.0802740 PubMedCrossRefPubMedCentralGoogle Scholar
  16. 16.
    Odink K, Cerletti N, Bruggen J, Clerc RG, Tarcsay L, Zwadlo G, Gerhards G, Schlegel R, Sorg C (1987) Two calcium-binding proteins in infiltrate macrophages of rheumatoid arthritis. Nature 330(6143):80–82. doi: 10.1038/330080a0 PubMedCrossRefGoogle Scholar
  17. 17.
    Vogl T, Tenbrock K, Ludwig S, Leukert N, Ehrhardt C, van Zoelen MA, Nacken W, Foell D, van der Poll T, Sorg C, Roth J (2007) Mrp8 and Mrp14 are endogenous activators of Toll-like receptor 4, promoting lethal, endotoxin-induced shock. Nat Med 13(9):1042–1049. doi: 10.1038/nm1638 PubMedCrossRefGoogle Scholar
  18. 18.
    Wang L, Chang EW, Wong SC, Ong SM, Chong DQ, Ling KL (2013) Increased myeloid-derived suppressor cells in gastric cancer correlate with cancer stage and plasma S100A8/A9 proinflammatory proteins. J Immunol 190(2):794–804. doi: 10.4049/jimmunol.1202088 PubMedCrossRefGoogle Scholar
  19. 19.
    Zhao F, Hoechst B, Duffy A, Gamrekelashvili J, Fioravanti S, Manns MP, Greten TF, Korangy F (2012) S100A9 a new marker for monocytic human myeloid-derived suppressor cells. Immunology 136(2):176–183. doi: 10.1111/j.1365-2567.2012.03566.x PubMedCrossRefPubMedCentralGoogle Scholar
  20. 20.
    Arai K, Takano S, Teratani T, Ito Y, Yamada T, Nozawa R (2008) S100A8 and S100A9 overexpression is associated with poor pathological parameters in invasive ductal carcinoma of the breast. Curr Cancer Drug Targets 8(4):243–252PubMedCrossRefGoogle Scholar
  21. 21.
    Ott HW, Lindner H, Sarg B, Mueller-Holzner E, Abendstein B, Bergant A, Fessler S, Schwaerzler P, Zeimet A, Marth C, Illmensee K (2003) Calgranulins in cystic fluid and serum from patients with ovarian carcinomas. Cancer Res 63(21):7507–7514PubMedGoogle Scholar
  22. 22.
    Vogl T, Gharibyan AL, Morozova-Roche LA (2012) Pro-inflammatory S100A8 and S100A9 proteins: self-assembly into multifunctional native and amyloid complexes. Int J Mol Sci 13(3):2893–2917. doi: 10.3390/ijms13032893 PubMedCrossRefPubMedCentralGoogle Scholar
  23. 23.
    Sinha P, Okoro C, Foell D, Freeze HH, Ostrand-Rosenberg S, Srikrishna G (2008) Proinflammatory S100 proteins regulate the accumulation of myeloid-derived suppressor cells. J Immunol 181(7):4666–4675PubMedCrossRefPubMedCentralGoogle Scholar
  24. 24.
    Bunt SK, Clements VK, Hanson EM, Sinha P, Ostrand-Rosenberg S (2009) Inflammation enhances myeloid-derived suppressor cell cross-talk by signaling through Toll-like receptor 4. J Leukoc Biol 85(6):996–1004. doi: 10.1189/jlb.0708446 PubMedCrossRefPubMedCentralGoogle Scholar
  25. 25.
    Yin C, Li H, Zhang B, Liu Y, Lu G, Lu S, Sun L, Qi Y, Li X, Chen W (2013) RAGE-binding S100A8/A9 promotes the migration and invasion of human breast cancer cells through actin polymerization and epithelial–mesenchymal transition. Breast Cancer Res Treat 142(2):297–309. doi: 10.1007/s10549-013-2737-1 PubMedCrossRefGoogle Scholar
  26. 26.
    Ye XZ, Yu SC, Bian XW (2010) Contribution of myeloid-derived suppressor cells to tumor-induced immune suppression, angiogenesis, invasion and metastasis. J Genet Genomics = Yi chuan xue bao 37(7):423–430. doi: 10.1016/S1673-8527(09)60061-8 CrossRefGoogle Scholar
  27. 27.
    Hiratsuka S, Watanabe A, Aburatani H, Maru Y (2006) Tumour-mediated upregulation of chemoattractants and recruitment of myeloid cells predetermines lung metastasis. Nat Cell Biol 8(12):1369–1375. doi: 10.1038/ncb1507 PubMedCrossRefGoogle Scholar
  28. 28.
    Zhang J, Liu J (2013) Tumor stroma as targets for cancer therapy. Pharmacol Ther 137(2):200–215. doi: 10.1016/j.pharmthera.2012.10.003 PubMedCrossRefPubMedCentralGoogle Scholar
  29. 29.
    Drews-Elger K, Brinkman JA, Miller P, Shah SH, Harrell JC, da Silva TG, Ao Z, Schlater A, Azzam DJ, Diehl K, Thomas D, Slingerland JM, Perou CM, Lippman ME, El-Ashry D (2014) Primary breast tumor-derived cellular models: characterization of tumorigenic, metastatic, and cancer-associated fibroblasts in dissociated tumor (DT) cultures. Breast Cancer Res Treat. doi: 10.1007/s10549-014-2887-9 Google Scholar
  30. 30.
    Simpson KD, Templeton DJ, Cross JV (2012) Macrophage migration inhibitory factor promotes tumor growth and metastasis by inducing myeloid-derived suppressor cells in the tumor microenvironment. J Immunol 189(12):5533–5540. doi: 10.4049/jimmunol.1201161 PubMedCrossRefPubMedCentralGoogle Scholar
  31. 31.
    Ichikawa M, Williams R, Wang L, Vogl T, Srikrishna G (2011) S100A8/A9 activate key genes and pathways in colon tumor progression. Mol Cancer Res 9(2):133–148. doi: 10.1158/1541-7786.MCR-10-0394 PubMedCrossRefPubMedCentralGoogle Scholar
  32. 32.
    Freund A, Chauveau C, Brouillet JP, Lucas A, Lacroix M, Licznar A, Vignon F, Lazennec G (2003) IL-8 expression and its possible relationship with estrogen-receptor-negative status of breast cancer cells. Oncogene 22(2):256–265. doi: 10.1038/sj.onc.1206113 PubMedCrossRefPubMedCentralGoogle Scholar
  33. 33.
    Pederson L, Winding B, Foged NT, Spelsberg TC, Oursler MJ (1999) Identification of breast cancer cell line-derived paracrine factors that stimulate osteoclast activity. Cancer Res 59(22):5849–5855PubMedGoogle Scholar
  34. 34.
    Verjans E, Noetzel E, Bektas N, Schutz AK, Lue H, Lennartz B, Hartmann A, Dahl E, Bernhagen J (2009) Dual role of macrophage migration inhibitory factor (MIF) in human breast cancer. BMC Cancer 9:230. doi: 10.1186/1471-2407-9-230 PubMedCrossRefPubMedCentralGoogle Scholar
  35. 35.
    McAllister SS, Gifford AM, Greiner AL, Kelleher SP, Saelzler MP, Ince TA, Reinhardt F, Harris LN, Hylander BL, Repasky EA, Weinberg RA (2008) Systemic endocrine instigation of indolent tumor growth requires osteopontin. Cell 133(6):994–1005. doi: 10.1016/j.cell.2008.04.045 PubMedCrossRefPubMedCentralGoogle Scholar
  36. 36.
    Kurebayashi J, Otsuki T, Kunisue H, Mikami Y, Tanaka K, Yamamoto S, Sonoo H (1999) Expression of vascular endothelial growth factor (VEGF) family members in breast cancer. Jpn J Cancer Res Gann 90(9):977–981CrossRefGoogle Scholar
  37. 37.
    Sorlie T, Perou CM, Tibshirani R, Aas T, Geisler S, Johnsen H, Hastie T, Eisen MB, van de Rijn M, Jeffrey SS, Thorsen T, Quist H, Matese JC, Brown PO, Botstein D, Lonning PE, Borresen-Dale AL (2001) Gene expression patterns of breast carcinomas distinguish tumor subclasses with clinical implications. Proc Natl Acad Sci USA 98(19):10869–10874. doi: 10.1073/pnas.191367098 PubMedCrossRefPubMedCentralGoogle Scholar
  38. 38.
    Karnoub AE, Dash AB, Vo AP, Sullivan A, Brooks MW, Bell GW, Richardson AL, Polyak K, Tubo R, Weinberg RA (2007) Mesenchymal stem cells within tumour stroma promote breast cancer metastasis. Nature 449(7162):557–563. doi: 10.1038/nature06188 PubMedCrossRefGoogle Scholar
  39. 39.
    Diaz-Cano SJ (2012) Tumor heterogeneity: mechanisms and bases for a reliable application of molecular marker design. Int J Mol Sci 13(2):1951–2011. doi: 10.3390/ijms13021951 PubMedCrossRefPubMedCentralGoogle Scholar
  40. 40.
    Zardavas D, Baselga J, Piccart M (2013) Emerging targeted agents in metastatic breast cancer. Nat Rev Clin Oncol 10(4):191–210. doi: 10.1038/nrclinonc.2013.29 PubMedCrossRefGoogle Scholar
  41. 41.
    Mahmoud SM, Paish EC, Powe DG, Macmillan RD, Grainge MJ, Lee AH, Ellis IO, Green AR (2011) Tumor-infiltrating CD8+ lymphocytes predict clinical outcome in breast cancer. J Clin Oncol Off J Am Soc Clin Oncol 29(15):1949–1955. doi: 10.1200/JCO.2010.30.5037 CrossRefGoogle Scholar
  42. 42.
    Condamine T, Gabrilovich DI (2011) Molecular mechanisms regulating myeloid-derived suppressor cell differentiation and function. Trends Immunol 32(1):19–25. doi: 10.1016/ PubMedCrossRefPubMedCentralGoogle Scholar
  43. 43.
    Sumida K, Wakita D, Narita Y, Masuko K, Terada S, Watanabe K, Satoh T, Kitamura H, Nishimura T (2012) Anti-IL-6 receptor mAb eliminates myeloid-derived suppressor cells and inhibits tumor growth by enhancing T-cell responses. Eur J Immunol 42(8):2060–2072. doi: 10.1002/eji.201142335 PubMedCrossRefGoogle Scholar
  44. 44.
    Morales JK, Kmieciak M, Knutson KL, Bear HD, Manjili MH (2010) GM-CSF is one of the main breast tumor-derived soluble factors involved in the differentiation of CD11b−Gr1− bone marrow progenitor cells into myeloid-derived suppressor cells. Breast Cancer Res Treat 123(1):39–49. doi: 10.1007/s10549-009-0622-8 PubMedCrossRefPubMedCentralGoogle Scholar
  45. 45.
    Xu X, Wang B, Ye C, Yao C, Lin Y, Huang X, Zhang Y, Wang S (2008) Overexpression of macrophage migration inhibitory factor induces angiogenesis in human breast cancer. Cancer Lett 261(2):147–157. doi: 10.1016/j.canlet.2007.11.028 PubMedCrossRefGoogle Scholar
  46. 46.
    Acharyya S, Oskarsson T, Vanharanta S, Malladi S, Kim J, Morris PG, Manova-Todorova K, Leversha M, Hogg N, Seshan VE, Norton L, Brogi E, Massague J (2012) A CXCL1 paracrine network links cancer chemoresistance and metastasis. Cell 150(1):165–178. doi: 10.1016/j.cell.2012.04.042 PubMedCrossRefPubMedCentralGoogle Scholar
  47. 47.
    Resetkova E, Reis-Filho JS, Jain RK, Mehta R, Thorat MA, Nakshatri H, Badve S (2010) Prognostic impact of ALDH1 in breast cancer: a story of stem cells and tumor microenvironment. Breast Cancer Res Treat 123(1):97–108. doi: 10.1007/s10549-009-0619-3 PubMedCrossRefGoogle Scholar
  48. 48.
    Nocito A, Kononen J, Kallioniemi OP, Sauter G (2001) Tissue microarrays (TMAs) for high-throughput molecular pathology research. Int J Cancer 94(1):1–5. doi: 10.1002/ijc.1385 PubMedCrossRefGoogle Scholar
  49. 49.
    Parker JS, Mullins M, Cheang MC, Leung S, Voduc D, Vickery T, Davies S, Fauron C, He X, Hu Z, Quackenbush JF, Stijleman IJ, Palazzo J, Marron JS, Nobel AB, Mardis E, Nielsen TO, Ellis MJ, Perou CM, Bernard PS (2009) Supervised risk predictor of breast cancer based on intrinsic subtypes. J Clin Oncol Off J Am Soc Clin Oncol 27(8):1160–1167. doi: 10.1200/JCO.2008.18.1370 CrossRefGoogle Scholar
  50. 50.
    Cooper A, van Doorninck J, Ji L, Russell D, Ladanyi M, Shimada H, Krailo M, Womer RB, Hsu JH, Thomas D, Triche TJ, Sposto R, Lawlor ER (2011) Ewing tumors that do not overexpress BMI-1 are a distinct molecular subclass with variant biology: a report from the Children’s Oncology Group. Clin Cancer Res Off J Am Assoc Cancer Res 17(1):56–66. doi: 10.1158/1078-0432.CCR-10-1417 CrossRefGoogle Scholar
  51. 51.
    Curtis C, Shah SP, Chin SF, Turashvili G, Rueda OM, Dunning MJ, Speed D, Lynch AG, Samarajiwa S, Yuan Y, Graf S, Ha G, Haffari G, Bashashati A, Russell R, McKinney S, Group M, Langerod A, Green A, Provenzano E, Wishart G, Pinder S, Watson P, Markowetz F, Murphy L, Ellis I, Purushotham A, Borresen-Dale AL, Brenton JD, Tavare S, Caldas C, Aparicio S (2012) The genomic and transcriptomic architecture of 2,000 breast tumours reveals novel subgroups. Nature 486(7403):346–352. doi: 10.1038/nature10983 PubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Katherine Drews-Elger
    • 1
    Email author
  • Elizabeth Iorns
    • 2
  • Alexandra Dias
    • 1
  • Philip Miller
    • 1
  • Toby M. Ward
    • 3
  • Sonja Dean
    • 1
  • Jennifer Clarke
    • 1
  • Adriana Campion-Flora
    • 4
  • Daniel Nava Rodrigues
    • 4
  • Jorge S. Reis-Filho
    • 5
  • James M. Rae
    • 6
  • Dafydd Thomas
    • 7
  • Deborah Berry
    • 8
  • Dorraya El-Ashry
    • 1
  • Marc E. Lippman
    • 1
  1. 1.Department of MedicineUniversity of Miami Miller School of MedicineMiamiUSA
  2. 2.Science ExchangePalo AltoUSA
  3. 3.Stanford Cancer InstituteStanford University School of MedicineStanfordUSA
  4. 4.Breakthrough Breast Cancer Research CenterThe Institute of Cancer ResearchLondonUK
  5. 5.Department of PathologyMemorial Sloan Kettering Cancer CenterNew YorkUSA
  6. 6.Breast Oncology Program, 6312 CCGCUniversity of Michigan Medical SchoolAnn ArborUSA
  7. 7.Department of PathologyUniversity of Michigan Medical SchoolAnn ArborUSA
  8. 8.Histopathology & Tissue Shared Resource, Lombardi Comprehensive Cancer CenterGeorgetown UniversityWashingtonUSA

Personalised recommendations