High-Content Assays for Hepatotoxicity Using Induced Pluripotent Stem Cell–Derived Cells

Abstract

Development of predictive in vitro assays for early toxicity evaluation is extremely important for improving the drug development process and reducing drug attrition rates during clinical development. High-content imaging-based in vitro toxicity assays are emerging as efficient tools for safety and efficacy testing to improve drug development efficiency. In this report we have used an induced pluripotent stem cell (iPSC)-derived hepatocyte cell model having a primary tissue-like phenotype, unlimited availability, and the potential to compare cells from different individuals. We examined a number of assays and phenotypic markers and developed automated screening methods for assessing multiparameter readouts of general and mechanism-specific hepatotoxicity. Endpoints assessed were cell viability, nuclear shape, average and integrated cell area, mitochondrial membrane potential, phospholipid accumulation, cytoskeleton integrity, and apoptosis. We assayed compounds with known mechanisms of toxicity and also evaluated a diverse hepatotoxicity library of 240 compounds. We conclude that high-content automated screening assays using iPSC-derived hepatocytes are feasible, provide information about mechanisms of toxicity, and can facilitate the safety assessment of drugs and chemicals.

Requirements for human iPS cell-derived hepatocytes as an alternative to primary human hepatocytes for assessing absorption, distribution, metabolism, excretion and toxicity of pharmaceuticals

“…The other is the large varia- tions between lots due to their different genetic and environmental backgrounds. HiPSC-derived hepatocytes have been anticipated to be used in in vitro ADMET studies as alternatives to PHHs because researchers are able to obtain the hiPSC-derived hepatocytes from the same origin repeatedly (Holmgren et al, 2014;Lu et al, 2015;Sirenko et al, 2014). PHHs were often used as controls in the studies of hiPSC-derived hepatocytes.…”

The Uses of Stem Cells in Toxicology Studies

“…hiPSC-kaynaklı hepatosit modelinin ilaç ve diğer kimyasalların güvenlik değerlendirmesini kolaylaştırabileceği ve toksisite mekanizmaları hakkında doğru bilgiler verebileceği sonucuna ulaşılmıştır [71]. hiPSC'ler birçok sitokrom (CYP) P450 genini eksprese etmektedir.…”

Toxicity testing and drug screening using iPSC-derived hepatocytes, cardiomyocytes, and neural cells

“…Recently, Sirenko et al (2014) developed imaging and analysis methods (high-content imaging-based assay using iPSC-derived hepatocyte cell model) providing tools for characterization of multiple toxicity phenotypes using live cells. Their results of testing commercially available hepatotoxic compounds showed significant promise for compound screening and safety evaluation during drug development.…”

“…Thus, animal models alone are unreliable for understanding D r a f t 4 There is growing impulse to move away from animal models for toxicity testing toward cell-based in vitro assays, predominantly in preliminary screening. Predictive in vitro cell-based assays should reduce or circumvent the dependence on animal models, simplify the assessment of candidate molecules and their metabolites, and facilitate evaluation of the mechanism of toxicity, kinetics, and dose-responses (Sirenko et al 2014). Combined with newer technology quantifying cell type-specific functions, human cell-based trials could be used to drive structure-activity relationship (SAR) to improve safety profiles.…”