Collagen Mineralization Decreases NK Cell‐Mediated Cytotoxicity of Breast Cancer Cells via Increased Glycocalyx Thickness

Skeletal metastasis is common in patients with advanced breast cancer and often caused by immune evasion of disseminated tumor cells (DTCs). In the skeleton, tumor cells not only disseminate to the bone marrow but also to osteogenic niches in which they interact with newly mineralizing bone extracellular matrix (ECM). However, it remains unclear how mineralization of collagen type I, the primary component of bone ECM, regulates tumor‐immune cell interactions. Here, a combination of synthetic bone matrix models with controlled mineral content, nanoscale optical imaging, and flow cytometry are utilized to evaluate how collagen type I mineralization affects the biochemical and biophysical properties of the tumor cell glycocalyx, a dense layer of glycosylated proteins and lipids decorating their cell surface. These results suggest that collagen mineralization upregulates mucin‐type O‐glycosylation and sialylation by tumor cells, which increases their glycocalyx thickness while enhancing resistance to attack by natural killer (NK) cells. These changes are functionally linked as treatment with a sialylation inhibitor decreased mineralization‐dependent glycocalyx thickness and made tumor cells more susceptible to NK cell attack. Together, these results suggest that interference with glycocalyx sialylation may represent a therapeutic strategy to enhance cancer immunotherapies targeting bone‐metastatic breast cancer. Skeletal metastasis in advanced breast cancer involves immune evasion by disseminated tumor cells. Using synthetic bone models with controlled mineral content and advanced imaging techniques, it is revealed that collagen type I mineralization in the bone matrix upregulates mucin‐type O‐glycosylation and sialylation in tumor cells, thickening their glycocalyx and enhancing resistance to natural killer cells.