Use of in vitro metabolomics in NRK cells to help predicting nephrotoxicity and differentiating the MoA of nephrotoxicants

• Published in: Toxicology letters 2021-12, Vol.353, p.43-59
• Main Authors: Birk, Barbara, Haake, Volker, Sperber, Saskia, Herold, Michael, Wallisch, Svenja K., Huener, Hans-Albrecht, Verlohner, Andreas, Amma, Meike M., Walk, Tilmann, Hernandez, Tzutzuy Ramirez, Hewitt, Nicola J., Kamp, Hennicke, van Ravenzwaay, Bennard
• Format: Article
• Language: English
• Subjects: Animals; Cell Line; Cell Survival - drug effects; Covalent protein binding; Drug-Related Side Effects and Adverse Reactions; In vitro; Kidney Diseases - chemically induced; Kidney Tubules - cytology; Lysosomal overload; Metabolomics; Mitochondrial DNA-interaction; Nephrotoxicity; NRK-52E cells; Rats
• ISSN: 0378-4274; 1879-3169
• DOI: 10.1016/j.toxlet.2021.09.011

[Display omitted] •In vitro metabolomics in kidney cells used to investigate nephrotoxicity MoAs.•A specific metabolic pattern was generated for different MoAs.•Almost all altered metabolites were lipids, indicating their role in nephrotoxicity.•Some specific key metabolites were identified.•A suitable tool for investigating the MoA of known and unknown nephrotoxicants. We describe a strategy using an in vitro metabolomics assay with tubular rat NRK-52E cells to investigate the Modes of Action (MoAs) of nephrotoxic compounds. Chemicals were selected according to their MoAs based on literature information: acetaminophen, 4-aminophenol and S-(trichlorovinyl-)L-cysteine (TCVC), (covalent protein binding); gentamycin, vancomycin, polymycin B and CdCl2 (lysosomal overload) and tenofovir and cidofovir (mitochondrial DNA-interaction). After treatment and harvesting of the cells, intracellular endogenous metabolites were quantified relative to vehicle control. Metabolite patterns were evaluated in a purely data-driven pattern generation process excluding published information. This strategy confirmed the assignment of the chemicals to the respective MoA except for TCVC and CdCl2. Finally, TCVC was defined as unidentified and CdCl2 was reclassified to the MoA “covalent protein binding”. Hierarchical cluster analysis of 58 distinct metabolites from the patterns enabled a clear visual separation of chemicals in each MoA. The assay reproducibility was very good and metabolic responses were consistent. These results support the use of metabolome analysis in NRK-52E cells as a suitable tool for understanding and investigating the MoA of nephrotoxicants. This assay could enable the early identification of nephrotoxic compounds and finally reduce animal testing.