A high-throughput metabolomics in vitro platform for the characterization of hepatotoxicity

• DOI: 10.1007/s10565-023-09809-6
• Publication Date: 2023-05-04T03:28:26Z
• Authors: Sabina Ramirez-Hi...; Barbara Birk; Philipp Ternes; Varun Giri; Volker Haake; Michael Herold; Franziska Maria Z...; Andreas Verlohner; Hans-Albrecht Huener; Hennicke Kamp; Peter Driemert; Robert Landsiedel; Elke Richling; Dorothee Funk-Weyer; Bennard van Raven...
• Keywords: Metabolome; High-Throughput Screening

Cell-based metabolomics provides multiparametric physiologically relevant readouts that can be highly advantageous for improved, biologically based decision making in early stages of compound development. Here, we present the development a 96-well plate LC-MS/MS-based targeted screening platform classification liver toxicity modes action (MoAs) HepG2 cells. Different parameters workflow (cell seeding density, passage number, cytotoxicity testing, sample preparation, metabolite extraction, analytical method, and data processing) were optimized standardized to increase efficiency testing platform. The applicability system was tested with seven substances known representative three different MoAs (peroxisome proliferation, enzyme induction, inhibition). Five concentrations per substance, aimed at covering complete dose-response curve, analyzed 221 uniquely identified metabolites measured, annotated, allocated 12 classes such as amino acids, carbohydrates, energy metabolism, nucleobases, vitamins cofactors, diverse lipid classes. Multivariate univariate analyses showed dose response metabolic effects, clear differentiation between resulted identification patterns specific each MoA. Key indicative both general mechanistic hepatotoxicity identified. method presented here offers multiparametric, mechanistic-based, cost-effective MoA sheds light into pathways involved toxicological mechanism. This assay implemented reliable improved safety assessment pipelines.