Nuclear Receptor-Mediated Gene Expression Changes in a Human Hepatic Micropatterned Coculture Model After Treatment with Hepatotoxic Compounds

Primary cultures of cryopreserved human hepatocytes have become the preferred model system to determine the potential of compounds to induce hepatic enzymes and transporters via nuclear receptor (NR) activation. Recently, it has been shown that cryopreserved hepatocytes retain viability, morphological integrity, and a more stable phenotype, including metabolic capacity, when maintained in micropatterned cocultures (MPCCs) with 3T3-J2 cells. However, no reports have been published demonstrating its potential as a surrogate for testing the NR activation potential of new compounds. The purpose of this study was to examine the maintenance and fidelity of key xenobiotic receptor pathways in human MPCCs, namely AhR, CAR, PXR, and PPARα. MPCCs from two separate batches of cryopreserved hepatocytes were treated with prototype NR activators and 17 compounds from the ToxCast and JRC datasets that are known to be hepatotoxic and/or activators of different NRs. Points of departure for chemical-induced changes in gene expression were calculated for NR-specific target genes after 24- and 72-hour treatments. Good concordance with induction of relevant target genes by the prototypical inducers was observed. Human MPCCs recapitulated outcomes between known activators and their respective receptor/targets, thus representing a reliable, robust, and reproducible surrogate model for the measurement of changes in NR-mediated gene expression.