Uncovering SOD3 and GPX4 as new targets of Benzo[α]pyrene-induced hepatotoxicity through Metabolomics and Chemical Proteomics
Yanwei Wang,
Jiahui Zhao,
Yipeng Xu,
Cimin Tao,
Jie Tong,
Yingjie Luo,
Yong Chen,
Xuesong Liu,
Tengfei Xu
Affiliations
Yanwei Wang
Key Laboratory of Advanced Drug Delivery Systems of Zhejiang Province, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, Zhejiang, China
Jiahui Zhao
Department of Geriatrics and Shenzhen Clinical Research Centre for Geriatrics, Shenzhen People’s Hospital (The Second Clinical Medical College, Jinan University, The First Affiliated Hospital, Southern University of Science and Technology), Shenzhen, Guangdong, 518020, China
Yipeng Xu
Department of Urology, Zhejiang Cancer Hospital, Hangzhou, Zhejiang, 310022, China
Cimin Tao
Key Laboratory of Advanced Drug Delivery Systems of Zhejiang Province, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, Zhejiang, China
Jie Tong
PET Center, Department of Radiology and Biomedical Imaging, Yale School of Medicine, New Haven, CT, 06520, USA
Yingjie Luo
Key Laboratory of Advanced Drug Delivery Systems of Zhejiang Province, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, Zhejiang, China; Cangnan County Qiushi Innovation Research Institute of Traditional Chinese Medicine, Wenzhou, Zhejiang, 325899, China
Yong Chen
Key Laboratory of Advanced Drug Delivery Systems of Zhejiang Province, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, Zhejiang, China; Cangnan County Qiushi Innovation Research Institute of Traditional Chinese Medicine, Wenzhou, Zhejiang, 325899, China
Xuesong Liu
Key Laboratory of Advanced Drug Delivery Systems of Zhejiang Province, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, Zhejiang, China; Cangnan County Qiushi Innovation Research Institute of Traditional Chinese Medicine, Wenzhou, Zhejiang, 325899, China
Tengfei Xu
Key Laboratory of Advanced Drug Delivery Systems of Zhejiang Province, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, Zhejiang, China; Cangnan County Qiushi Innovation Research Institute of Traditional Chinese Medicine, Wenzhou, Zhejiang, 325899, China; Corresponding author. Key Laboratory of Advanced Drug Delivery Systems of Zhejiang Province, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, Zhejiang, China.
Benzo[α]pyrene (Bap) is recognized as a ubiquitous environmental pollutant among the polycyclic aromatic hydrocarbons (PAHs) class. Previous studies have shown that the hepatotoxicity of Bap is mainly caused by its metabolites, although it remains unclear whether Bap itself induces such damage. This study integrated metabolomics and chemical proteomics approaches to comprehensively identify the potential target proteins affected by Bap in liver cells. The results from the metabolomics showed that the significant changed metabolites were related with cellular redox homeostasis. CEllular Thermal Shift Assay (CETSA) showed that Bap induced protein thermal displacement of superoxide dismutase 3 (SOD3) and glutathione peroxidase 4 (GPX4), which are closely related to oxidative homeostasis. Further validation through in vitro CETSA and drug affinity response target stability (DARTS) revealed that Bap directly affected the stability of SOD3 and GPX4 proteins. The binding affinities of Bap to the potential target proteins were further evaluated using molecular docking, while the isothermal titration calorimetry (ITC) interaction measurements indicated nanomolar-level Kd values. Importantly, we found that Bap weakened the antioxidant capacity by destroying the activities of SOD3 and GPX4, which provided a new understanding of the mechanism of hepatotoxicity induced by Bap. Moreover, our provided workflow integrating metabolomics and label-free chemical proteomics, can be regarded as a practical way to identify the targets and inter-mechanisms for the various environmental compounds.
