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Updated: Dec 18, 2025
Epithelial Cell Repopulation and Preparation of Rodent Extracellular Matrix Scaffolds for Renal Tissue Development
Published on: August 10, 2015
New phenotypic cytotoxicity assay for ROS-inducing compounds using rat renal epithelial cells
Noriko Uchiyama1, Tomoya Yukawa1, Yvonne P Dragan1
1Global Drug Safety Research Evaluation, Takeda Pharmaceutical Company Ltd., Pharmaceutical Research Division, 40 Landsdowne Street, Cambridge, MA, 02139, United States.
Abstract:
An important mechanism of chemical toxicity is the induction of oxidative stress through the production of excess reactive oxygen species (ROS). In this study, we show that the level of drug-induced ROS production between NRK52E and HepG2 cells is significantly different for several marketed drugs and a number of Takeda's internal proprietary compounds. Nifedipine, a calcium channel blocker and the initial focus of the study, was demonstrated to promote in vitro ROS production and a decrease in cell viability in NRK52E cells but not HepG2 cells. ROS production after nifedipine treatment was inhibited by a NOX inhibitor (GKT136901) but not the mitochondrial NADH dehydrogenase inhibitor, rotenone, suggesting that nifedipine decreases NRK52E cell viability primarily through a NOX-mediated pathway. To understand the breadth of NOX-mediated ROS production, 12 commercially available compounds that are structurally and/or pharmacologically related to nifedipine as well as 172 internal Takeda candidate drugs, were also evaluated against these two cell types. Over 15 % of compounds not cytotoxic to HepG2 cells (below 50 μM) were cytotoxic to NRK52E cells. Our results suggest that a combination of cell viability data from both NRK52E and HepG2 cells was superior for the prediction of in vivo toxicity findings when compared to use of only one cell line. Further, the NRK52E cell viability assay is a good predictor of NOX-mediated ROS production and can be used as a follow up assay following a negative HepG2 response to aid in the selection of suitable compounds for in vivo toxicity studies.
Insights
Drug toxicity varies by cell type, with NRK52E cells showing sensitivity to compounds not affecting HepG2 cells. Combining cell viability data improves in vivo toxicity prediction, highlighting NRK52E assays for NOX-mediated oxidative stress.
Area of Science:
- Toxicology
- Cell Biology
- Pharmacology
Background:
- Chemical toxicity often involves oxidative stress from excess reactive oxygen species (ROS).
- Drug-induced ROS production can differ significantly between cell lines.
- NRK52E and HepG2 cells exhibit differential responses to various compounds.
Purpose of the Study:
- To investigate differential drug-induced ROS production and cytotoxicity between NRK52E and HepG2 cells.
- To elucidate the mechanism of nifedipine-induced toxicity in NRK52E cells.
- To evaluate the utility of combined cell line data for predicting in vivo toxicity.
Main Methods:
- Assessed in vitro ROS production and cell viability in NRK52E and HepG2 cells following drug treatment.
- Utilized NOX inhibitor (GKT136901) and mitochondrial inhibitor (rotenone) to probe nifedipine's mechanism.
- Screened marketed drugs and Takeda proprietary compounds for differential cytotoxicity.
Main Results:
- Nifedipine induced ROS and decreased viability in NRK52E cells, but not HepG2 cells, via a NOX-mediated pathway.
- Over 15% of compounds non-cytotoxic to HepG2 cells were cytotoxic to NRK52E cells.
- Combined cell viability data from both cell lines improved prediction of in vivo toxicity.
Conclusions:
- NRK52E cell viability assays are predictive of NOX-mediated ROS production.
- Using both NRK52E and HepG2 cell data enhances the prediction of in vivo drug toxicity.
- This approach aids in selecting compounds with better safety profiles for further studies.
