Toxicity and underlying lipidomic alterations generated by a mixture of water disinfection byproducts in human lung cells

Complex mixtures of disinfection by-products (DBPs) are present in disinfected waters, but their mixture toxicity has been rarely described. Apart from ingestion, DBP exposure can occur through inhalation, which may lead to respiratory effects in highly exposed individuals. However, the underlying biological mechanisms have yet to be elucidated. This study aimed to investigate the toxicity of a mixture of 10 DBPs, including haloacetic acids and haloaromatics, on human alveolar A549 cells by assessing their cytotoxicity, genotoxicity, and impact on the cell lipidome. A DBP mixture up to 50 μM slightly reduced cell viability, induced the generation of reactive oxygen species (ROS) up to 3.5-fold, and increased the frequency of micronuclei formation. Exposure to 50 μM DBP mixture led to a significant accumulation of triacylglycerides (TGs) and a decrease of diacylglycerides (DGs) and phosphatidylcholines (PCs) in A549 cells. Lipidomic profiling of extracellular vesicles (EVs) released in the culture medium revealed a marked increase in cholesterol esters (CEs), sphingomyelins (SMs), and other membrane lipids. Overall, these alterations in the lipidome of cells and EVs may indicate a disruption of lipid homeostasis, potentially contributing to the respiratory effects associated with DBP exposure.