Granulocyte Colony Stimulating Factor Expression in Breast Cancer and Its Association with Carbonic Anhydrase IX and Immune Checkpoints

Abstract

Purpose: Granulocyte colony-stimulating factor (G-CSF) and hypoxia modulate the tumour immune microenvironment. In model systems, hypoxia-induced carbonic anhydrase IX (CAIX) has been associated with G-CSF and immune responses, including M2 polarization of macrophages. We investigated whether these associations exist in human breast cancer specimens, their relation to breast cancer subtypes, and clinical outcome. Methods: Using validated protocols and prespecified scoring methodology, G-CSF expression on carcinoma cells and CD163 expression on tumour-associated macrophages were assayed by immunohistochemistry and applied to a tissue microarray series of 2960 primary excision specimens linked to clinicopathologic, biomarker, and outcome data. Results: G-CSFhigh expression showed a significant positive association with ER negativity, HER2 positivity, presence of CD163+ M2 macrophages, and CAIX expression. In univariate analysis, G-CSFhigh phenotype was associated with improved survival in non-luminal cases, although the CAIX+ subset had a significantly adverse prognosis. A significant positive association was observed between immune checkpoint biomarkers on tumour-infiltrating lymphocytes and both G-CSF- and CAIX-expressing carcinoma cells. Immune checkpoint biomarkers correlated significantly with favourable prognosis in G-CSFhigh/non-luminal cases independent of standard clinicopathological features. Conclusions: The prognostic associations linking G-CSF to immune biomarkers and CAIX strongly support their immunomodulatory roles in the tumour microenvironment.

Prognostic Significance of CSF-1R Expression in Early Invasive Breast Cancer

“…We and others have previously shown that CD163+ M2 macrophages are associated with poor prognosis in breast cancer [ 25 , 54 ]. We performed an exploratory analysis to examine the relationship of carcinoma cells expressing CSF-1R in the context of tumor-associated macrophages stratified by ER expression.…”

“…The median follow-up of the patients is 12.5 years. This cohort has been comprehensively profiled for several key biomarkers, [ 50 ] including those related to immune-oncology, i.e., stromal tumor-infiltrating lymphocytes (assessed on hematoxylin- and eosin-stained slides), CD8, FOXP3, TIM3, LAG-3, PD-1 (on intraepithelial tumor-infiltrating lymphocytes (iTILs)), PD-L1 (on carcinoma cells), and CD163 (on macrophages), and assessed by immunohistochemistry [ 51 , 52 , 53 , 54 , 55 ]. The patient population of the BC Cancer cohort has been split into internal training and validation sets as described previously [ 56 ].…”

“…Images from these slides are available for public access via the website of the Genetic Pathology Evaluation Center ( (accessed on 27 September 2021)). The scoring methodologies for the immune biomarkers have been described previously [ 51 , 52 , 53 , 54 , 55 ].…”

The immune microenvironment and relation to outcome in patients with advanced breast cancer treated with docetaxel with or without gemcitabine

“…38 Supplementary, CD163, a macrophage marker, was assessed for predictive capacity. 39 Analyses were carried out on the whole cohort, and as a preselected subgroup analysis on the non-luminal breast cancers, a category combining the basal-like and HER2enriched (HER2E) PAM50 subtypes based on published research showing these two subtypes to be similar in their tumor-immune microenvironment. 7,32 Tests for heterogeneity of prognostic significance in luminal subtypes (luminal A and luminal B) versus non-luminal subtypes were also performed.…”

“…48 The cutoffs for CD163 expressing tumor associated macrophages were also adjusted from studies in our previous cohort and were set to <56 vs ≥56. 39 PD-L1 scoring was performed according to manufacturer's guidelines as area of PD-L1 positive TILs out of total tumor area. 49 A cutoff of ≥1% for positive expression was used when relevant, as this is the commonly used cutoff for clinical trials and treatment indications.…”