TY - GEN A1 - Rosellini, Matteo A1 - Schulze, Alicia A1 - Omer, Ejlal A. A1 - Ali, Nadeen T. A1 - Marini, Federico A1 - Küpper, Jan-Heiner A1 - Efferth, Thomas T1 - The effect of plastic-related compounds on transcriptome-wide gene expression on CYP2C19-overexpressing HepG2 cells T2 - Molecules N2 - In recent years, plastic and especially microplastic in the oceans have caused huge problems to marine flora and fauna. Recently, such particles have also been detected in blood, breast milk, and placenta, underlining their ability to enter the human body, presumably via the food chain and other yet-unknown mechanisms. In addition, plastic contains plasticizers, antioxidants, or lubricants, whose impact on human health is also under investigation. At the cellular level, the most important enzymes involved in the metabolism of xenobiotic compounds are the cytochrome P450 monooxygenases (CYPs). Despite their extensive characterization in the maintenance of cellular balance, their interactions with plastic and related products are unexplored. In this study, the possible interactions between several plastic-related compounds and one of the most important cytochromes, CYP2C19, were analyzed. By applying virtual compound screening and molecular docking to more than 1000 commercially available plastic-related compounds, we identified candidates that are likely to interact with this protein. A growth inhibition assay confirmed their cytotoxic activity on a CYP2C19-transfected hepatic cell line. Subsequently, we studied the effect of the selected compounds on the transcriptome-wide gene expression level by conducting RNA sequencing. Three candidate molecules were identified, i.e., 2,2′-methylene bis(6-tert-butyl-4-methylphenol), 1,1-bis(3,5-di-tert-butyl-2-hydroxyphenyl) ethane, and 2,2′-methylene bis(6-cyclohexyl-4-methylphenol)), which bound with a high affinity to CYP2C19 in silico. They exerted a profound cytotoxicity in vitro and interacted with several metabolic pathways, of which the ‘cholesterol biosynthesis process’ was the most affected. In addition, other affected pathways involved mitosis, DNA replication, and inflammation, suggesting an increase in hepatotoxicity. These results indicate that plastic-related compounds could damage the liver by affecting several molecular pathways. KW - cytotoxicity KW - ecotoxicity KW - hepatotoxicity KW - microplastic KW - RNA sequencing Y1 - 2023 UR - https://www.mdpi.com/1420-3049/28/16/5952 U6 - https://doi.org/10.3390/molecules28165952 SN - 1420-3049 VL - 28 IS - 16 ER - TY - GEN A1 - Rosellini, Matteo A1 - Omer, Ejlal A. A1 - Schulze, Alicia A1 - Ali, Nadeen T. A1 - Boulos, Joelle C. A1 - Marini, Federico A1 - Küpper, Jan-Heiner A1 - Efferth, Thomas T1 - Impact of plastic-related compounds on the gene expression signature of HepG2 cells transfected with CYP3A4 T2 - Archives of Toxicology N2 - The presence of plastic and microplastic within the oceans as well as in marine flora and fauna have caused a multitude of problems that have been the topic of numerous investigations for many years. However, their impact on human health remains largely unknown. Such plastic and microplastic particles have been detected in blood and placenta, underlining their ability to enter the human body. Plastics also contain other compounds, such as plasticizers, antioxidants, or dyes, whose impact on human health is currently being studied. Critical enzymes within the metabolism of endogenous molecules, especially of xenobiotics, are the cytochrome P450 monooxygenases (CYPs). Although their importance in maintaining cellular balance has been confirmed, their interactions with plastics and related products are poorly understood. In this study, the possible relationship between different plastic-related compounds and CYP3A4 as one of the most important CYPs was analyzed using hepatic cells overexpressing this enzyme. Beginning with virtual compound screening and molecular docking of more than 1000 plastic-related compounds, several candidates were identified to interact with CYP3A4. In a second step, RNA-sequencing was used to study in detail the transcriptome-wide gene expression levels affected by the selected compounds. Three candidate molecules ((2,2′-methylenebis(6- tert -butyl-4-methylphenol), 1,1-bis(3,5-di- tert -butyl-2-hydroxyphenyl)ethane, and 2,2′-methylenebis(6-cyclohexyl-4-methylphenol)) had an excellent binding affinity to CYP3A4 in-silico as well as cytotoxic effects and interactions with several metabolic pathways in-vitro. We identified common pathways influenced by all three selected plastic-related compounds. In particular, the suppression of pathways related to mitosis and ‘DNA-templated DNA replication’ which were confirmed by cell cycle analysis and single-cell gel electrophoresis. Furthermore, several mis-regulated metabolic and inflammation-related pathways were identified, suggesting the induction of hepatotoxicity at different levels. These findings imply that these compounds may cause liver problems subsequently affecting the entire organism. KW - Cytotoxicity KW - Ecotoxicity KW - Environmental pollution KW - Hepatotoxicity KW - Marine pollution KW - Microplastic KW - RNA-sequencing KW - Biological Sciences Y1 - 2024 U6 - https://doi.org/10.1007/s00204-023-03648-4 SN - 0340-5761 SN - 1432-0738 VL - 98 IS - 2 SP - 525 EP - 536 PB - Springer Berlin Heidelberg ER -