Quantification of distinct behavioromes of anti-tumor and pro-tumor human neutrophil subtypes in a microphysiological system 3611

Abstract Description Neutrophils can either inhibit or promote tumor progression. Studies in mice suggest that polarizing neutrophils from a pro-tumor “N2” phenotype to an anti-tumor “N1” phenotype could be a promising therapeutic strategy. Understanding the behavioral differences between N1 and N2 human neutrophil subtypes in the solid tumor tissue is important for developing novel cancer immunotherapy. In this study, we engineered a human cell-based microphysiological system to quantify both real-time and end-point behaviors of LPS, IFN-γ, and IFN-β-polarized N1-like and TGF-β-polarized N2-like neutrophils during their interactions with pancreatic tumor spheroids in a 3D collagen matrix. N1-like neutrophils showed higher levels of pro-inflammatory hallmarks including ROS production, IL-8 secretion, and NET release and 2D cancer cytotoxicity than N2-like neutrophils. N1-like neutrophils were less motile in response to tumor spheroids, engaged in longer contact with tumor spheroids, and infiltrated greater into tumor spheroids than N2-like neutrophils. N1-like neutrophils also attenuated tumor progression in terms of invasion, proliferation, and epithelial-mesenchymal transition compared to N2-like neutrophils, although tumor apoptosis was not affected. Our microphysiological system reveals the distinct “behavioromes” of anti-tumor and pro-tumor human neutrophils in pancreatic cancer, which may inform the design of novel cancer immunotherapies aiming to modulate specific neutrophil behaviors. Funding Sources This work was supported by start-up funds and NSF CBET CAREER Award (2240094) from Caroline N. Jones. Shuai Shao was supported by the Jonsson Family Bioengineering Fellowship 2023-2024 and the Bioengineering Convergent Grant 2024 from UT Dallas. Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)