Spatial Metrics of Interaction between CD163-Positive Macrophages and Cancer Cells and Progression-Free Survival in Chemo-Treated Breast Cancer.

Abstract

Simple SummaryA majority of breast cancer deaths are caused by aggressive molecular subtypes that are at high risk of progression. Patients with high-risk breast cancer commonly receive first-line systemic chemotherapy. Chemotherapy exerts direct cytotoxic effects on proliferating cancer cells. In addition, significant effects of chemotherapy are mediated through immune-boosting anti-cancer mechanisms that counteract immunosuppressive tumor-associated macrophages (TAMs). The aim of our study was to evaluate the potential prognostic value of the prevalence and the spatial localization of CD TAMs in tumor tissue from breast cancer patients treated with chemotherapy after surgery. We developed a novel algorithm that identifies CD TAMs in an objective manner and quantifies spatial interactions between CD TAMs and cancer cells using distance-based metrics. Our results demonstrate that close spatial proximity of CD TAMs to cancer cells and the average number of CD cells either directly adjacent to or within communicating distance of each cancer cell are independent predictors of unfavorable prognosis in breast cancer.Tumor-associated macrophages (TAMs) promote progression of breast cancer and other solid malignancies via immunosuppressive, pro-angiogenic and pro-metastatic effects. Tumor-promoting TAMs tend to express M2-like macrophage markers, including CD163. Histopathological assessments suggest that the density of CD163-positive TAMs within the tumor microenvironment is associated with reduced efficacy of chemotherapy and unfavorable prognosis. However, previous analyses have required research-oriented pathologists to visually enumerate CD TAMs, which is both laborious and subjective and hampers clinical implementation. Objective, operator-independent image analysis methods to quantify TAM-associated information are needed. In addition, since M2-like TAMs exert local effects on cancer cells through direct juxtacrine cell-to-cell interactions, paracrine signaling, and metabolic factors, we hypothesized that spatial metrics of adjacency of M2-like TAMs to breast cancer cells will have further information value. Immunofluorescence histo-cytometry of CD TAMs was performed retrospectively on tumor microarrays of 443 cases of invasive breast cancer from patients who subsequently received adjuvant chemotherapy. An objective and automated algorithm was developed to phenotype CD TAMs and calculate their density within the tumor stroma and derive several spatial metrics of interaction with cancer cells. Shorter progression-free survival was associated with a high density of CD TAMs, shorter median cancer-to-CD nearest neighbor distance, and a high number of either directly adjacent CD TAMs (within juxtacrine proximity <12 μm to cancer cells) or communicating CD TAMs (within paracrine communication distance <250 μm to cancer cells) after multivariable adjustment for clinical and pathological risk factors and correction for optimistic bias due to dichotomization.