Xenobiotica-metabolizing enzyme induction potential of chemicals in animal studies: NanoString nCounter gene expression
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TOXICOKINETICS AND METABOLISM
Xenobiotica‑metabolizing enzyme induction potential of chemicals in animal studies: NanoString nCounter gene expression and peptide group‑specific immunoaffinity as accelerated and economical substitutions for enzyme activity determinations? Brandy Riffle1 · Franz Oesch2 · Annika Heckmanns3 · Eric Fabian3 · Mao Wang1 · Anita Samuga1 · Peifeng Ren1 · Helen Hammer4 · Felix Schmidt5 · Oliver Pötz4 · Bennard van Ravenzwaay3 · Robert Landsiedel3 Received: 13 February 2020 / Accepted: 4 May 2020 © Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Xenobiotica-metabolizing enzyme (XME) induction is a relevant biological/biochemical process vital to understanding the toxicological profile of xenobiotics. Early recognition of XME induction potential of compounds under development is therefore important, yet its determination by traditional XME activity measurements is time consuming and cost intensive. A proof-of-principle study was therefore designed due to the advent of faster and less cost-intensive methods for determination of enzyme protein and transcript levels to determine whether two such methods may substitute for traditional measurement of XME activity determinations. The results of the study show that determination of enzyme protein levels by peptide group-specific immunoaffinity enrichment/MS and/or determination of gene expression by NanoString nCounter may serve as substitutes for traditional evaluation methodology and/or as an early predictor of potential changes in liver enzymes. In this study, changes of XME activity by the known standard XME inducers phenobarbital, beta-naphthoflavone and Aroclor 1254 were demonstrated by these two methods. To investigate the applicability of these methods to demonstrate XME-inducing activity of an unknown, TS was also examined and found to be an XME inducer. More specifically, TS was found to be a phenobarbital-type inducer (likely mediated by CAR rather than PXR as nuclear receptor), but not due to Ah receptor-mediated or antioxidant response element-mediated beta-naphthoflavone-type induction. The results for TS were confirmed via enzymatic activity measurements. The results of the present study demonstrate the potential applicability of NanoString nCounter mRNA quantitation and peptide group-specific immunoaffinity enrichment/MS protein quantitation for predicting compounds under development to be inducers of liver XME activity. Keywords Xenobiotica-metabolizing enzyme induction · Peptide group-specific immunoaffinity enrichment/MS · NanoString nCounter Brandy Riffle and Franz Oesch contributed equally. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00204-020-02777-4) contains supplementary material, which is available to authorized users. * Robert Landsiedel [email protected] 1
BASF Corporation, 26 Davis Drive, Research Triangle Park, NC 27709, USA
2
Institute of Toxicology, Johannes Gutenberg University of Mainz, Rheinblick 21, 55263 Wackernheim, G
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