S1: The Resolution of Concentration Response: High Throughput Transcriptomics (S1500+) Paired with In Vitro Liver Models Reveal the...

...Importance of Cell Culture Configuration on Hepatocyte Functionality & the Resolving Power to Distinguish ‘Safer’ Case Study Analogues & Liver Injury Compounds

S1: The Resolution of Concentration Response:  High Throughput Transcriptomics, OpenTox USA 2017

Stephen S. Ferguson 






Stephen S. Ferguson, Sreenivasa C. Ramaiahgari, Trey Saddler, Julie Rice, Paul Dunlap, Stephanie Smith-Roe, Kristine Witt, Nisha Sipes, Alex Merrick, Michael J. DeVito, Richard S. Paules, Scott Auerbach


As part of Phase III of the Tox21 Program, we are developing improved approaches to investigate chemical safety/toxicity potential in humans. Initial efforts have focused on developing and integrating organotypic in vitro liver screening models (2D & 3D) and high dimensional assay systems (i.e., high throughput transcriptomics (HTT), morphological imaging, cell health assays). In this presentation, we describe our findings with a panel of 24 compounds designed to probe the impact of cell culture configuration (HepaRG cells) on cellular differentiation (i.e., xenobiotic metabolism, hepatic receptor pathway functionality, modeling of metabolically-activated biological responses (i.e., toxicity), and the power of broad-range concentration response evaluations to discriminate ‘therapeutic’ vs. ‘off-target’ or toxic responses to chemical exposures. The study highlights the major limitations of proliferating cell culture models, even with highly differentiated cells such as HepaRG, to mimic tissue-like responses to compounds (i.e., phenobarbital, fenofibric acid, chenodeoxycholic acid, rosiglitazone), the impact of proliferating status on baseline xenobiotic metabolism competence, the potential to model metabolically-activated toxicity (e.g., cyclophosphamide, troglitazone, acetaminophen, aflatoxin B1), and the promising utility to distinguish ‘case study’ liver injury compounds from less hepatotoxic analogues via increased propensities for increased/pleiotropic transcript perturbations. The approach, coupled with quantitative in vitro to in vivo extrapolation methods, provides a data-rich and cost-effective solution to rapidly survey biological response space, in a potency-driven framework, to more effectively evaluate chemical-induced biological perturbations in humans.