Plasma Profile of Immune Determinants Predicts Pathological Complete Response in Locally Advanced Breast Cancer Patients: A Pilot Study

  • Author Footnotes
    # R.M., C.C., J.W., and R.O. contributed equally to this study.
    Rosalba Miceli
    # R.M., C.C., J.W., and R.O. contributed equally to this study.
    Clinical Epidemiology and Trial Organization Unit, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, Italy
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  • Author Footnotes
    # R.M., C.C., J.W., and R.O. contributed equally to this study.
    Chunmei Cao
    # R.M., C.C., J.W., and R.O. contributed equally to this study.
    Department of Breast Surgery, Fudan University Shanghai Cancer Center, Shanghai, People's Republic of China
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  • Nicolai N. Maolanon
    Department of Breast Surgery, Fudan University Shanghai Cancer Center, Shanghai, People's Republic of China

    Department of Chemistry, University of Copenhagen, Copenhagen, Denmark
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  • Roland Jacobs
    Department of Rheumatology and Clinical Immunology, Hannover Medical School, Hannover, Germany
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  • Author Footnotes
    # R.M., C.C., J.W., and R.O. contributed equally to this study.
    Jiong Wu
    # R.M., C.C., J.W., and R.O. contributed equally to this study.
    Department of Breast Surgery, Fudan University Shanghai Cancer Center, Shanghai, People's Republic of China
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  • Author Footnotes
    # R.M., C.C., J.W., and R.O. contributed equally to this study.
    Rosaria Orlandi
    Address for correspondence: Rosaria Orlandi, Molecular Targeting Unit, Fondazione IRCCS Istituto Nazionale dei Tumori, Via Amadeo 42, 20133 Milan, Italy
    # R.M., C.C., J.W., and R.O. contributed equally to this study.
    Molecular Targeting Unit, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, Italy
    Search for articles by this author
  • Author Footnotes
    # R.M., C.C., J.W., and R.O. contributed equally to this study.



      Complex interactions between cancer and the immune system have an impact on disease progression and therapeutic response. Our objective was to evaluate whether circulating immune-related determinants are associated with pathological complete response (pCR) in patients with locally advanced breast cancer (LABC) subjected to neoadjuvant chemotherapy (NACT).

      Patients and Methods

      Luminex technology was used to profile 22 cytokines, 10 chemokines, FGF2, PDGF-BB, VEGF, and Ca15-3/Ca125 glycoforms. Measurements were performed alongside standard hematological determinations on pretreatment plasma samples from 151 patients including 41 cases with pCR assessed following RECIST criteria.


      Random Forest model analysis selected platelets, eotaxin, IFN-γ, IP10, and TGFβ2 as significant predictors of pCR. These immune-related features were combined into a quantitative score predictive of pCR. In patients who scored 0 or 1, none had pCR; the pCR frequency increased in relation to the score value (23.5%, 41.2%, and 78.6%, in score groups 2, 3, and 4, respectively). At multivariable logistic analysis, the pCR score was highly significant (odds ratio = 3.15 per unit increment; CI: 1.85-5.38; P < .0001); among clinical covariates (age, menopausal status, tumor stage, IHC subtype, Ki-67, CA15.3, and CA125), only Ki-67 was statistically significant (P = .013).


      This explorative study aimed to lay the conceptual and practical foundation that a distinctive pattern of the immune determinant blood signature at diagnosis of LABC significantly correlates with the patient's response to NACT and provides the groundwork for larger studies that could lead to a minimally invasive tool for personalized medicine.



      BC (breast cancer), BMI (body max index), CI (confidence interval), ER (estrogen receptor), HER2 (human epidermal growth factor receptor 2), HR (hormonal receptors), IHC (immunohistochemistry), OR (odds ratio), pCR (pathological complete response), PR (progesterone receptor), RECIST (Response Evaluation Criteria in Solid Tumors), TNBC (triple negative breast cancer)
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        • Kong X
        • Moran MS
        • Zhang N
        • Haffty B
        • Yang Q.
        Meta-analysis confirms achieving pathological complete response after neoadjuvant chemotherapy predicts favourable prognosis for breast cancer patients.
        Eur J Cancer. 2011; 47: 2084-2090
        • Cortazar P
        • Zhang L
        • Untch M
        • et al.
        Pathological complete response and long-term clinical benefit in breast cancer: the CTNeoBC pooled analysis.
        Lancet. 2014; 384 (Erratum in: Lancet. 2019 Mar 9;393:986. PMID: 24529560): 164-172
        • von Minckwitz G
        • Untch M
        • Blohmer JU
        • et al.
        Definition and impact of pathologic complete response on prognosis after neoadjuvant chemotherapy in various intrinsic breast cancer subtypes.
        J Clin Oncol. 2012; 30: 1796-1804
        • Houssami N
        • Macaskill P
        • von Minckwitz G
        • Marinovich ML
        • Mamounas E.
        Meta-analysis of the association of breast cancer subtype and pathologic complete response to neoadjuvant chemotherapy.
        Eur J Cancer. 2012; 48: 3342-3354
        • Rouzier R
        • Perou CM
        • Symmans WF
        • et al.
        Breast cancer molecular subtypes respond differently to preoperative chemotherapy.
        Clin Cancer Res. 2005; 11: 5678-5685
        • Masuda H
        • Baggerly KA
        • Wang Y
        • et al.
        Differential response to neoadjuvant chemotherapy among 7 triple-negative breast cancer molecular subtypes.
        Clin Cancer Res. 2013; 19: 5533-5540
        • Tudorica A
        • Oh KY
        • Chui SY
        • et al.
        Early prediction and evaluation of breast cancer response to neoadjuvant chemotherapy using quantitative DCE-MRI.
        Transl Oncol. 2016; 9: 8-17
        • Schreiber RD
        • Old LJ
        • Smyth MJ.
        Cancer immunoediting: integrating immunity's roles in cancer suppression and promotion.
        Science. 2011; 331: 1565-1570
        • Fridman WH
        • Zitvogel L
        • Sautès-Fridman C
        • Kroemer G.
        The immune contexture in cancer prognosis and treatment.
        Nat Rev Clin Oncol. 2017; 14: 717-734
        • Cimino-Mathews A
        • Foote JB
        • Emens LA.
        Immune targeting in breast cancer.
        Oncology (Williston Park). 2015; 29 (PubMed PMID: 25979549): 375-385
        • Denkert C
        • Loibl S
        • Noske A
        • et al.
        Tumor-associated lymphocytes as an independent predictor of response to neoadjuvant chemotherapy in breast cancer.
        J Clin Oncol. 2010; 28 (PMID:19917869): 105-113
        • Verma C
        • Kaewkangsadan V
        • Eremin JM
        • et al.
        Natural killer (NK) cell profiles in blood and tumour in women with large and locally advanced breast cancer (LLABC) and their contribution to a pathologicalcomplete response (PCR) in the tumour following neoadjuvant chemotherapy (NAC): differential restoration of blood profiles by NAC and surgery.
        J Transl Med. 2015; 13: 180
        • Kuznetsov HS
        • Marsh T
        • Markens BA
        • et al.
        Identification of luminal breast cancers that establish a tumor-supportive macroenvironment defined by proangiogenic platelets and bone marrow-derived cells.
        Cancer Discov. 2012; 2: 1150-1165
        • McAllister SS
        • Weinberg RA.
        The tumour-induced systemic environment as a critical regulator of cancer progression and metastasis.
        Nat Cell Biol. 2014; 16: 717-727
        • Spitzer MH
        • Carmi Y
        • Reticker-Flynn NE
        • et al.
        Systemic immunity is required for effective cancer immunotherapy.
        Cell. 2017; 168 (e15): 487-502
        • Orlandi R
        • De Bortoli M
        • Ciniselli CM
        • et al.
        Hepcidin and ferritin blood level as noninvasive tools for predicting breast cancer.
        Ann Oncol. 2014; 25: 352-357
        • Dumeaux V
        • Ursini-Siegel J
        • Flatberg A
        • et al.
        Peripheral blood cells inform on the presence of breast cancer: a population-based case-control study.
        Int J Cancer. 2015; 136: 656-667
        • Dumeaux V
        • Fjukstad B
        • Fjosne HE
        • et al.
        Interactions between the tumor and the blood systemic response of breast cancer patients.
        PLoS Comput Biol. 2017; 13e1005680
        • Kawaguchi K
        • Suzuki E
        • Yamaguchi A
        • Yamamoto M
        • Morita S
        • Toi M.
        Altered expression of major immune regulatory molecules in peripheral blood immune cells associated with breast cancer.
        Breast Cancer. 2017; 24: 111-120
        • Wang S
        • Zhang Z
        • Fang F
        • Gao X
        • Sun W
        • Liu H.
        The neutrophil/lymphocyte ratio is an independent prognostic indicator in patients with bone metastasis.
        Oncol Lett. 2011; 2: 735-740
        • Seruga B
        • Zhang H
        • Bernstein LJ
        • Tannock IF.
        Cytokines and their relationship to the symptoms and outcome of cancer.
        Nat Rev Cancer. 2008; 8: 887-899
        • Carlsson A
        • Wingren C
        • Kristensson M
        • et al.
        Molecular serum portraits in patients with primary breast cancer predict the development of distant metastases.
        Proc Natl Acad Sci U S A. 2011; 108: 14252-14257
        • Hong CC
        • Yao S
        • McCann SE
        • et al.
        Pretreatment levels of circulating Th1 and Th2 cytokines, and their ratios, are associated with ER-negative and triple negative breast cancers.
        Breast Cancer Res Treat. 2013; 139: 477-488
        • Allavena P
        • Germano G
        • Marchesi F
        • Mantovani A.
        Chemokines in cancer related inflammation.
        Exp Cell Res. 2011; 317: 664-673
        • Lippitz BE.
        Cytokine patterns in patients with cancer: a systematic review.
        Lancet Oncol. 2013; 14: e218-e228
        • Therasse P
        • Arbuck SG
        • Eisenhauer EA
        • et al.
        New guidelines to evaluate the response to treatment in solid tumors. European Organization for Research and Treatment of Cancer, National Cancer Institute of the United States, National Cancer Institute of Canada.
        J Natl Cancer Inst. 2000; 92 (PubMed PMID: 10655437): 205-216
        • Chen K
        • Gentry-Maharaj A
        • Burnell M
        • et al.
        Microarray glycoprofiling of CA125 improves differential diagnosis of ovarian cancer.
        J Proteome Res. 2013; 12: 1408-1418
        • Vignali DA.
        Multiplexed particle-based flow cytometric assays.
        J Immunol Methods. 2000; 243: 243-255
        • Freeman GH
        • Halton JH.
        Note on an exact treatment of contingency, goodness of fit and other problems of significance.
        Biometrika. 1951; 38 (PMID: 14848119): 141-149
      1. Breiman L. (2001) Random forests. Mach Learn. 45, 5-32.

        • Altmann A
        • Toloşi L
        • Sander O
        • Lengauer T.
        Permutation importance: a corrected feature importance measure.
        Bioinformatics. 2010; 26: 1340-1347
        • Durrleman S
        • Simon R.
        Flexible regression models with cubic splines.
        Stat Med. 1989; 8: 551-561
        • Tian L
        • Tibshirani R.
        Adaptive index models for marker-based risk stratification.
        Biostatistics. 2011; 12: 68-86
        • Hosmer DW
        • Lemeshow S.
        Applied Logistic Regression.
        Wiley, New York2000
        • Heinze G
        • Schemper M.
        A solution to the problem of separation in logistic regression.
        Stat Med. 2002; 21: 2409-2419
        • Harrell Jr, FE
        • Lee KL
        • Mark DB.
        Multivariable prognostic models: issues in developing models, evaluating assumptions and adequacy, and measuring and reducing errors.
        Stat Med. 1996; 15: 361-387<361::AID-SIM168>3.0.CO;2-4
        • Efron B.
        • Tibshirani R.J.
        An Introduction to the Bootstrap.
        Chapman and Hall, New York1993
        • Core Team R
        R: A Language and Environment for Statistical Computing.
        R Foundation for Statistical Computing, Vienna, Austria2016 (URL Last accessed February 2018)
        • Huang X
        • Dugo M
        • Callari M
        • et al.
        Molecular portrait of breast cancer in China reveals comprehensive transcriptomic likeness to Caucasian breast cancer and low prevalence of luminal A subtype.
        Cancer Med. 2015; 4: 1016-1030
        • Duffy MJ.
        Tumor markers in clinical practice: a review focusing on common solid cancers.
        Med Princ Pract. 2013; 22: 4-11
        • Delmar P
        • Irl C
        • Tian L.
        Innovative methods for the identification of predictive biomarker signatures in oncology: application to bevacizumab.
        Contemp Clin Trials Commun. 2017; 5: 107-115
        • Mancuso ME
        • Santagostino E.
        Platelets: much more than bricks in a breached wall.
        Br J Haematol. 2017; 178 (Epub 2017Apr 17): 209-219
        • Wiesner T
        • Bugl S
        • Mayer F
        • Hartmann JT
        • Kopp HG.
        Differential changes in platelet VEGF, Tsp, CXCL12, and CXCL4 in patients with metastatic cancer.
        Clin Exp Metastasis. 2010; 27: 141-149
        • Best MG
        • Sol N
        • Kooi I
        • et al.
        RNA-Seq of tumor-educated platelets enables blood-based pan-cancer, multiclass, and molecular pathway cancer diagnostics.
        Cancer Cell. 2015; 28: 666-676
        • Liu S
        • Fang J
        • Jiao D
        • Liu Z.
        Elevated platelet count predicts poor prognosis in breast cancer patients with supraclavicular lymph node metastasis.
        Cancer Manag Res. 2020; 12 (Published 2020 Jul 20): 6069-6075
        • Drabsch Y
        • ten Dijke P.
        TGF-β signaling in breast cancer cell invasion and bone metastasis.
        J Mammary Gland Biol Neoplasia. 2011; 16: 97-108
        • Heidegger I
        • Höfer J
        • Luger M
        • et al.
        Is Eotaxin-1 a serum and urinary biomarker for prostate cancer detection and recurrence?.
        Prostate. 2015; 75: 1904-1909
        • Wang L
        • Simons DL
        • Lu X
        • et al.
        Breast cancer induces systemic immune changes on cytokine signaling in peripheral blood monocytes and lymphocytes.
        EBioMedicine. 2020; 52102631
        • Müller C
        • Tufa DM
        • Chatterjee D
        • Mühlradt PF
        • Schmidt RE
        • Jacobs R.
        The TLR-2/TLR-6 agonist macrophage-activating lipopeptide-2 augments human NK cell cytotoxicity when PGE2 production by monocytes is inhibited by a COX-2 blocker.
        Cancer Immunol Immunother. 2015; 64: 1175-1184
        • Jafarzadeh A
        • Fooladseresht H
        • Nemati M
        • Assadollahi Z
        • Sheikhi A
        • Ghaderi A.
        Higher circulating levels of chemokine CXCL10 in patients with breast cancer: evaluation of the influences of tumor stage and chemokine gene polymorphism.
        Cancer Biomark. 2016; 16: 545-554
        • Narita D
        • Seclaman E
        • Anghel A
        • et al.
        Altered levels of plasma chemokines in breast cancer and their association with clinical and pathological characteristics.
        Neoplasma. 2016; 63: 141-149
        • Jabeen S
        • Espinoza JA
        • Torland LA
        • et al.
        Noninvasive profiling of serum cytokines in breast cancer patients and clinicopathological characteristics.
        Oncoimmunology. 2018; 8e1537691
        • Espinoza JA
        • Jabeen S
        • Batra R
        • et al.
        Cytokine profiling of tumor interstitial fluid of the breast and its relationship with lymphocyte infiltration and clinicopathological characteristics.
        Oncoimmunology. 2016; 5e1248015
      2. Jabeen S, Zucknick M, Nome M, et al. Serum cytokine levels in breast cancer patients during neoadjuvant treatment with bevacizumab. Oncoimmunology. 2018;7:e1457598. doi:10.1080/2162402X.2018.1457598.