Identification of toxicity pathway of diesel particulate matter using AOP of PPARγ inactivation leading to pulmonary fibrosis

Diesel particulate matter (DPM), a major subset of urban fine particulate matter (PM2.5), raises huge concerns for human health and has therefore been classified as a group 1 carcinogen by the International Agency for Research on Cancer (IARC). However, as DPM is a complex mixture of various chemicals, understanding of DPM's toxicity mechanism remains limited. As the major exposure route of DPM is through inhalation, we herein investigated its toxicity mechanism based on the Adverse Outcome Pathway (AOP) of pulmonary fibrosis, which we previously submitted to AOPWiki as AOP ID 206 (AOP206). We first screened whether individual chemicals in DPM have the potential to exert their toxicity through AOP206 by using the ToxCast database and deep learning models approach, then confirmed this by examining whether DPM as a mixture alters the expression of the molecular initiating event (MIE) and key events (KEs) of AOP206. For identifying the activeness of the component chemicals of DPM, we used 24 ToxCast assays potentially related to AOP206 and deep learning models based on these assays, which were identified and developed in our previous study. Of the 100 individual chemicals in DPM, 34 were active in PPARγ (MIE)-related assay, of which 17 were active in one or more KEs. To further identify whether individual chemicals in DPM are related to the MIE of AOP206, we performed molecular docking simulation on PPARγ for the chemicals showing activeness. Benzo[e]pyrene, benzo[a]pyrene and other related chemicals were the most likely to bind to PPARγ. In in vitro experiments, PPARγ activity increased with exposure of the DPM mixture, and the protein expression of PPARγ (MIE), and fibronectin (AO) also tended to be increased. Overall, we have demonstrated that AOP206 can be applied to identify the toxicity pathway of DPM. Further, we suggest that applying the AOP approach using ToxCast and deep learning models is useful for identifying potential toxicity pathways of chemical mixtures, such as DPM, by determining the activity of individual chemicals.