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Evaluation of In Vitro Biotransformation Using HepaRG Cells to Improve High-Throughput Chemical Hazard Prediction: A Toxicogenomics Analysis (SOT)
Citation:
Franzosa, J., J. Jack, P. Kothiya, D. Filer, J. Liu, J. Bonzo, S. Ferguson, I. Shah, A. Richard, R. Thomas, J. Wambaugh, AND K. Houck. Evaluation of In Vitro Biotransformation Using HepaRG Cells to Improve High-Throughput Chemical Hazard Prediction: A Toxicogenomics Analysis (SOT). Presented at 2015 Annual SOT Meeting, San Diego, CA, March 22 - 26, 2015. https://doi.org/10.23645/epacomptox.5178868
Impact/Purpose:
Present poster at SOT annual meeting in San Diego, CA on March 24, 2015
Description:
The US EPA’s ToxCast program has generated a wealth of data in >600 in vitro assayson a library of 1060 environmentally relevant chemicals and failed pharmaceuticals to facilitate hazard identification. An inherent criticism of many in vitro-based strategies is the inability of assays to detect in vivo relevant activities in cells lacking biotransformation capacity. To address this potential limitation, weused an in vitro liver toxicogenomics approach to explore gene-specific perturbations elicited by ToxCast library chemicals in metabolically-competent HepaRG™ cells. Here, cells were treated in eight-point concentration-response for 48 hours in a 96-well plate. The expression of 93 genes was profiled via qPCR using Fluidigm 96.96 dynamic arrays. To leverage the xenobiotic metabolism competency of the cells, these results were combined with morphologic assessments and lactate dehydrogenase assay evaluation of cytotoxicity to characterize effects on biological processes and disease states, including: cell proliferation, survival and death; nuclear receptor (NR)-mediated metabolism and transport; oxidative stress; and steatosis. Genes regulated by xenobiotic-sensing NRs (CAR, PXR, PPARa, FXR) as well as AhR, were induced upon exposure to positive controls and CYP-inducing chemicals, such as polycyclic aromatic hydrocarbons and organophosphate pesticides. Potential molecular initiating events were identified to further adverse outcome pathway elucidation for target organ (liver) effects often hallmark of systemic toxicity. Furthermore, these data complement existing ToxCast data by facilitating identification and investigation into false positives and negatives to improve chemical hazard prediction and strengthen the utility of alternative methods to support human risk assessment decisions. This abstract does not necessarily reflect EPA policy
URLs/Downloads:
DOI: Evaluation of In Vitro Biotransformation Using HepaRG Cells to Improve High-Throughput Chemical Hazard Prediction: A Toxicogenomics Analysis (SOT)