Revealing consensus gene pathways associated with respiratory functions and disrupted by PM2.5 nitrate exposure at...

Nitrate is a major pollutant component in ambient PM_2.5. It is known that chronic exposure to PM_2.5 NO₃^- damages respiratory functions. We aim to explore the underlying toxicological mechanism at single cell resolution.

We systematically conducted exposure experiments on forty C57BL/6 mice, assessed respiratory functions, and profiled lung transcriptome. . Afterward, we estimated the cell type compositions from RNA-seq data using deconvolution analysis. The genes and pathways associated with respiratory function and dysregulated by to PM_2.5 NO₃^- exposure were characterized at bulk-tissue and single-cell resolution.

PM_2.5 NO₃^- exposure did not significantly modify the cell type composition in lung, but profoundly altered the gene expression within each cell type. At ambient concentration (22 μg/m³), exposure significantly (FDR<10%) altered 95 genes' expression. Among the genes associated with respiratory functions, a large fraction (74.6-91.7%) were significantly perturbed by PM_2.5 NO₃^- exposure. For example, among the 764 genes associated with peak expiratory flow (PEF), 608 (79.6%) were affected by exposure (p = 1.92e-345). Pathways known to play role in lung disease pathogenesis, including circadian rhythms, sphingolipid metabolism, immune response and lysosome, were found significantly associated with respiratory functions and disrupted by PM_2.5 NO₃^- exposure.

This study extended our knowledge of PM_2.5 NO₃^- exposure's effect to the levels of lung gene expression, pathways, lung cell type composition and cell specific transcriptome. At single cell resolution, we provided insights in toxicological mechanism of PM_2.5 NO₃^- exposure and subsequent pulmonary disease risks.