Discovering a predictive metabolic signature of drug-induced structural cardiotoxicity in cardiac microtissues.

Abstract

Improved prediction of drug-induced structural cardiotoxicity is required to reduce attrition driven by cardiac safety concerns in drug discovery. Omics measurements are well suited to this need, offering the potential to discover molecular signatures associated with toxicological endpoints. In addition, untargeted metabolomics can simultaneously measure xenobiotic fate within the test system. We present an extensive metabolomics study to discover a predictive metabolic signature of drug-induced structural cardiotoxicity. A human-relevant in vitro cardiac model, cardiac microtissues, were exposed to twelve xenobiotics (eight clinically labelled structural cardiotoxins and four non-cardiotoxic pharmaceuticals), each at two concentrations, for 6, 24, and 48 h. The measurements were made by direct-infusion and liquid-chromatography mass spectrometry from intracellular polar and lipid extracts, and spent culture medium, respectively. Data were used to quantify levels, and reveal the metabolic fate of the xenobiotics, and to simultaneously explore their effects on the cardiac microtissues. Xenobiotic quantification revealed free concentrations to be typically lower than nominal values, whilst discovery of xenobiotic-related features evidenced the biotransformation capacity of the microtissues. Both common and condition-specific effects of the xenobiotics on the intracellular metabolome, lipidome, and metabolic footprint were discovered. Moreover, metabolic signatures with capacity to predict structural cardiotoxicity were revealed. These included features representing several ceramides, energy metabolism intermediates, e.g. creatine, purine-related metabolites, and markers of oxidative stress, e.g. glutathione.