Neutrophils alter the metabolic profile of a coculture model of triple-negative breast cancer and stromal cells

Abstract Background Breast cancer is one of the most common cancer types. Triple-negative breast cancer (TNBC), one of the most lethal breast cancer subtypes, is characterized by substantial neutrophil accumulation in the tumor microenvironment (TME), although the impact of neutrophils on disease progression remains unclear. Methods To model interactions between cancer cells and stromal cells, we used a coculture system combining a TNB Ccell line with breast fibroblasts and exposed these cultures to neutrophils isolated from healthy donors. Results Coculture generated a pro-inflammatory environment, with increased levels of pro-inflammatory cytokines resembling cytokine profiles reported in cancer patients. Coculture-conditioned medium attracted neutrophils and activated them to produce reactive oxygen species (ROS). Coculture also altered metabolic activity, as measured by the Seahorse Extracellular Flux Assay, compared with the corresponding monocultures. Conclusions Adding neutrophils to the coculture increased mitochondrial activity, suggesting that neutrophils may promote a metabolic shift in the coculture system: Together, our results support further investigation of neutrophil-driven changes in energy metabolism in TNBC microenvironments.