title: Relevance of Tumor-Infiltrating Immune Cell Composition and Functionality for Disease Outcome in Breast Cancer.
abstract: Not all breast cancer patients benefit from neoadjuvant or adjuvant therapy, resulting in considerable undertreatment or overtreatment. New insights into the role of tumor-infiltrating immune cells suggest that their composition, as well as their functionality, might serve as a biomarker to enable optimal patient selection for current systemic therapies and upcoming treatment options such as immunotherapy.
  
  
  
  We performed several complementary unbiased in silico analyses on gene expression profiles of 7270 unrelated tumor samples of nonmetastatic breast cancer patients with known clinical follow-up. CIBERSORT was used to estimate the fraction of 22 immune cell types to study their relations with pathological complete response (pCR), disease-free survival (DFS), and overall survival (OS). In addition, we used four previously reported immune gene signatures and a CD8+ T-cell exhaustion signature to assess their relationships with breast cancer outcome. Multivariable binary logistic regression and multivariable Cox regression were used to assess the association of immune cell-type fractions and immune signatures with pCR and DFS/OS, respectively.
  
  
  
  Increased fraction of regulatory T-cells in human epidermal growth factor receptor 2 (HER2)-positive tumors was associated with a lower pCR rate (odds ratio [OR] = 0.15, 95% confidence interval [CI] = 0.03 to 0.69), as well as shorter DFS (hazard ratio [HR] = 3.13, 95% CI = 1.23 to 7.98) and OS (HR = 7.69, 95% CI = 3.43 to 17.23). A higher fraction of M0 macrophages in estrogen receptor (ER)-positive tumors was associated with worse DFS (HR = 1.66, 95% CI = 1.18 to 2.33) and, in ER-positive/HER2-negative tumors, with worse OS (HR = 1.71, 95% CI = 1.12 to 2.61). Increased fractions of γδ T-cells in all breast cancer patients related to a higher pCR rate (OR = 1.55, 95% CI = 1.01 to 2.38), prolonged DFS (HR = 0.68, 95% CI = 0.48 to 0.98), and, in HER2-positive tumors, with prolonged OS (HR = 0.27, 95% CI = 0.10 to 0.73). A higher fraction of activated mast cells was associated with worse DFS (HR = 5.85, 95% CI = 2.20 to 15.54) and OS (HR = 5.33, 95% CI = 2.04 to 13.91) in HER2-positive tumors. The composition of relevant immune cell types frequently differed per breast cancer subtype. Furthermore, a high CD8+ T-cell exhaustion signature score was associated with shortened DFS in patients with ER-positive tumors regardless of HER2 status (HR = 1.80, 95% CI = 1.07 to 3.04).
  
  
  
  The main hypothesis generated in our unbiased in silico approach is that a multitude of immune cells are related to treatment response and outcome in breast cancer.
authors.affiliation: Affiliations of authors: Department of Medical Oncology, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands (RDB, MATMvV, EGEdV, CPS, RSNF); Department of Medical Oncology and Breast Cancer Translational Research Laboratory, Institut Jules Bordet, Université Libre de Bruxelles, Brussels, Belgium (CS, MJPG); Division of Immunology, Netherlands Cancer Institute, Amsterdam, the Netherlands (JBAGH).
authors.forename: Rico D
authors.initials: RD
authors.lastname: Bense
authors.affiliation: Affiliations of authors: Department of Medical Oncology, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands (RDB, MATMvV, EGEdV, CPS, RSNF); Department of Medical Oncology and Breast Cancer Translational Research Laboratory, Institut Jules Bordet, Université Libre de Bruxelles, Brussels, Belgium (CS, MJPG); Division of Immunology, Netherlands Cancer Institute, Amsterdam, the Netherlands (JBAGH).
authors.forename: Christos
authors.initials: C
authors.lastname: Sotiriou
authors.affiliation: Affiliations of authors: Department of Medical Oncology, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands (RDB, MATMvV, EGEdV, CPS, RSNF); Department of Medical Oncology and Breast Cancer Translational Research Laboratory, Institut Jules Bordet, Université Libre de Bruxelles, Brussels, Belgium (CS, MJPG); Division of Immunology, Netherlands Cancer Institute, Amsterdam, the Netherlands (JBAGH).
authors.forename: Martine J
authors.initials: MJ
authors.lastname: Piccart-Gebhart
authors.affiliation: Affiliations of authors: Department of Medical Oncology, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands (RDB, MATMvV, EGEdV, CPS, RSNF); Department of Medical Oncology and Breast Cancer Translational Research Laboratory, Institut Jules Bordet, Université Libre de Bruxelles, Brussels, Belgium (CS, MJPG); Division of Immunology, Netherlands Cancer Institute, Amsterdam, the Netherlands (JBAGH).
authors.forename: John B A G
authors.initials: JBAG
authors.lastname: Haanen
authors.affiliation: Affiliations of authors: Department of Medical Oncology, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands (RDB, MATMvV, EGEdV, CPS, RSNF); Department of Medical Oncology and Breast Cancer Translational Research Laboratory, Institut Jules Bordet, Université Libre de Bruxelles, Brussels, Belgium (CS, MJPG); Division of Immunology, Netherlands Cancer Institute, Amsterdam, the Netherlands (JBAGH).
authors.forename: Marcel A T M
authors.initials: MATM
authors.lastname: van Vugt
authors.affiliation: Affiliations of authors: Department of Medical Oncology, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands (RDB, MATMvV, EGEdV, CPS, RSNF); Department of Medical Oncology and Breast Cancer Translational Research Laboratory, Institut Jules Bordet, Université Libre de Bruxelles, Brussels, Belgium (CS, MJPG); Division of Immunology, Netherlands Cancer Institute, Amsterdam, the Netherlands (JBAGH).
authors.forename: Elisabeth G E
authors.initials: EGE
authors.lastname: de Vries
authors.affiliation: Affiliations of authors: Department of Medical Oncology, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands (RDB, MATMvV, EGEdV, CPS, RSNF); Department of Medical Oncology and Breast Cancer Translational Research Laboratory, Institut Jules Bordet, Université Libre de Bruxelles, Brussels, Belgium (CS, MJPG); Division of Immunology, Netherlands Cancer Institute, Amsterdam, the Netherlands (JBAGH).
authors.forename: Carolien P
authors.initials: CP
authors.lastname: Schröder
authors.affiliation: Affiliations of authors: Department of Medical Oncology, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands (RDB, MATMvV, EGEdV, CPS, RSNF); Department of Medical Oncology and Breast Cancer Translational Research Laboratory, Institut Jules Bordet, Université Libre de Bruxelles, Brussels, Belgium (CS, MJPG); Division of Immunology, Netherlands Cancer Institute, Amsterdam, the Netherlands (JBAGH).
authors.forename: Rudolf S N
authors.initials: RSN
authors.lastname: Fehrmann
mesh_terms: D001943:Breast Neoplasms / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D003198:Computer Simulation; D018572:Disease-Free Survival; D005260:Female; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000276:immunology*; D016246:Lymphocytes, Tumor-Infiltrating / Q000737:chemistry; D008264:Macrophages / Q000276:immunology*; D008407:Mast Cells / Q000276:immunology*; D008875:Middle Aged; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000032:analysis; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000032:analysis; D011960:Receptors, Estrogen / Q000032:analysis; D015996:Survival Rate; D050378:T-Lymphocytes, Regulatory / Q000276:immunology; D059467:Transcriptome
chemical_list: D018719:Erb-b2 Receptor Tyrosine Kinases; D016692:Receptors, Antigen, T-Cell, gamma-delta; D011960:Receptors, Estrogen; C508053:ERBB2 protein, human