Multi-Omics Assessment of Puff Volume-Mediated Salivary Biomarkers of Metal Exposure and Oxidative Injury Associated with Electronic Nicotine Delivery Systems.
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BACKGROUND: Since their inception, electronic nicotine delivery systems (ENDS) have gained increasing popularity, sparking a vaping epidemic among adolescents in the US and globally. Several ENDS safety concerns have emerged as device features and formats that contribute to heavy metal exposure and toxicity continue to evolve and outpace regulatory efforts. OBJECTIVES: Our objective was to integrate ENDS emission profiles with salivary proteome and metabolome data to characterize exposure factors that may influence adverse vaping-mediated health outcomes. METHODS: A total of 56 participants (38 exclusive ENDS users and 18 non-ENDS users) were recruited. A subset of 15 exclusive ENDS users completed puffing topography assessments to obtain individual vaping behavior patterns using each participant's ENDS device. Inductively coupled plasma mass spectrometry was used to determine the metal content of emissions (12 ENDS devices) generated using a programmable ENDS aerosol generation system and saliva (15 exclusive ENDS users and 5 non-ENDS users). Saliva samples from 10 exclusive ENDS users and 5 non-ENDS users were analyzed for proteomic, metabolomic, inflammatory, and oxidative stress/damage biomarkers. RESULTS: A linear puff volume-dependent increase in particle emissions and heavy metals was observed in ENDS aerosols and saliva of exclusive ENDS users. Elevated puff volume-dependent levels of salivary cytokines, including tumor necrosis factor-alpha (TNFα), interleukin-1 beta (IL-1β), and IL-6, were observed alongside the oxidative damage indicators malondialdehyde (MDA), superoxide dismutase (SOD), and 8-hydroxy-2'-deoxyguanosine (8-OHdG). Proteome-metabolome network analysis showed a higher risk of potentially developing neurological and respiratory diseases in ENDS users compared with non-ENDS users. Integrated salivary proteome-metabolome-metallome network analysis further demonstrated that heavy metals were associated with proteomic and metabolomic perturbations, with notable alterations in inflammatory response, immune function, and disease-related pathways. DISCUSSION: A significant correlation between heavy metals, cytokines, and oxidative stress markers reveals a potential role of vaping behavior in exposure to metals and changes in markers associated with DNA damage and inflammation. Our study demonstrates the importance of characterizing vaping behavior and puffing topography when examining the human health implications of ENDS use. https://doi.org/10.1289/EHP14321.