Assessing pulmonary response to TNF-α in human precision-cut lung slices using single-cell RNA sequencing 4294

Abstract Description Respiratory infections pose significant health challenges, particularly for high-risk groups. Understanding host responses to respiratory pathogens may open new avenues for therapeutic strategies. Human-relevant models are essential for studying host-targeting therapies that modulate immune responses and enable rapid, adaptable treatments. This study characterizes a human precision cut lung slice (hPCLS) model to investigate early, cell-type-specific inflammatory responses seen during pulmonary infections. hPCLSs were exposed to TNF-α, a pro-inflammatory cytokine induced during infection, and cellular and molecular changes were assessed using single-cell RNA sequencing. The data identified T cells, B cells, NK cells, macrophages, as well as epithelial, fibroblast, and endothelial populations. Notably, following a 24-hour exposure to TNF-α, minimal cell proliferation was observed across these populations. Macrophages exhibited the highest TNF-α receptor expression and upregulation of myeloid migration and chemotaxis genes. TNF-α exposure elevated chemokine expression (CXCL17, CCL2) in endothelial, macrophage, and epithelial cells. Additionally, epithelial cells upregulated apoptosis-related genes, indicating a damage response. These findings highlight the hPCLS model’s ability to capture early, cell-specific lung responses to inflammation, reinforcing its value as a preclinical tool to study host-pathogen interactions and inform future therapeutic strategies. Funding Sources This work was performed under the auspices of the U.S. Department of Energy by Lawrence Livermore National Laboratory under Contract DE-AC52-07NA27344. Funding was provided by internal Lawrence Livermore National Laboratory Directed Research and Development funds (24-ERD-025) to N.R.H. LLNL-ABS-2001042. Topic Categories Cellular Adhesion, Migration, and Inflammation (CAM)