Abstract 2806: Progression of ductal carcinoma in situ (DCIS), is it in the immune microenvironment

Introduction: DCIS is a non-obligate precursor to invasive breast cancer (IBC). DCIS patients are treated similarly to breast cancer with surgery, often followed by radiotherapy and/or endocrine treatment. However, most DCIS lesions will never progress to IBC, indicating that overdiagnosis and overtreatment exists. DCIS lesions show variable amounts of immune cells, particularly in the periductal stroma. Immune escape might be a critical step for transition from DCIS to IBC. We aim to identify factors within the immune microenvironment of DCIS lesions that distinguish harmless from potentially hazardous DCIS. Methods: A case-control study is being conducted consisting of women with pure DCIS diagnosed between 1989-2005 with median follow-up of 12 years, treated with breast conserving surgery only. Cases are defined as women with DCIS developing subsequent ipsilateral breast cancer (iIBC), controls as women with DCIS without subsequent iIBC. Multispectral immunohistochemical imaging was performed on primary DCIS lesions, aiming at detection of CD20+ B-cells, CD8+ T-cells, CD3+ T-cells, CD3+Foxp3+ regulatory T-cells, and CD68+ macrophages. Density of immune cell subsets in cells/mm2, immune cell ratios and spatial relationships were calculated for 27 cases and 28 controls. These immune cell related factors were correlated to outcome and integrated with RNAseq data of pure microdissected DCIS. We performed gene set enrichment analysis on the correlation between DCIS gene expression and density of immune cell types with sample permutation (flexgsea R package). Results: Stromal lymphocyte, B-cell, CD8+ T-cell, regulatory T-cell and macrophage density did not significantly differ between cases and controls. Immune cell composition (CD8+ T-cell/lymphocyte, CD8+ T-cell/CD3+Foxp3+ regulatory T-cell and CD20+/lymphocyte ratio) and fraction of regulatory T-cells in close proximity of a CD8+ T-cell did not differ between cases and controls. We find a negative association between stromal B-cell density and DCIS gene expression of estrogen receptor (ESR1) targets. Higher stromal T-cell density was associated with proliferation and expression of genes characteristic for luminal B and basal-like subtypes. Furthermore, higher density of specific immune cell subsets within the DCIS compartment was associated with several immune and cancer pathways. Conclusion: A first set of analyzed DCIS cases (n=27) and controls (n=28) show no significant differences regarding immune cell