Systematic transcriptome-based comparison of cellular adaptive stress response activation networks in hepatic stem cell-derived progeny and primary human hepatocytes

Bas Ter Braak,Marije Niemeijer,Ruben Boon,Céline Parmentier,Audrey Baze,Lysiane Richert,Suzanna Huppelschoten,Steven Wink,Catherine Verfaillie,Bob Van De Water

doi:10.1016/j.tiv.2021.105107

Bas Ter Braak, Marije Niemeijer + Show 8 more

Open Access

https://doi.org/10.1016/j.tiv.2021.105107

Copy DOI

Abstract

Various adaptive cellular stress response pathways are critical in the pathophysiology of liver disease and drug-induced liver injury. Human-induced pluripotent stem cell (hiPSC)-derived hepatocyte-like cells (HLCs) provide a promising tool to study cellular stress response pathways, but in this context there is limited insight on how HLCs compare to other in vitro liver models. Here, we systematically compared the transcriptomic profiles upon chemical activation in HLCs, hiPSC, primary human hepatocytes (PHH) and HepG2 liver cancer cells. We used targeted RNA-sequencing to map concentration transcriptional response using benchmark concentration modeling for the various stress responses in the different test systems. We found that HLCs are very sensitive towards oxidative stress and inflammation conditions as corresponding genes were activated at over 3 fold lower concentrations of the corresponding pathway inducing compounds as compared to PHH. PHH were the most sensitive model when studying UPR related effects. Due to the non-proliferative nature of PHH and HLCs, these do not pose a good/sensitive model to pick up DNA damage responses, while hiPSC and HepG2 were more sensitive in these conditions. We envision that this study contributes to a better understanding on how HLCs can contribute to the assessment of cell physiological stress response activation to predict hepatotoxic events.

Highlights

Drug-induced liver injury (DILI) is one of the most frequent causes for drug withdrawal and the leading cause for compound attrition in drug development (Kullak-Ublick et al, 2017)
In the last phase of the differentiation protocol, hepatocyte growth factor (HGF) was added to the liver differentiation medium (LDM) which resulted in cells with a typical cuboidal morphology (Acikgoz et al, 2013), that stained positive for anti-alpha trypsin (AAT) and hepatocyte nuclear factor 4 alpha (HNF4α) (Fig. 1c)
Given that chemicals cause cell injury that triggers cellular stress response pathways, a question that has yet to be answered is how Human-induced pluripotent stem cell (hiPSC)-hepatic progeny compares to primary human hepatocytes (PHH) in the context of cellular stress response activation

Summary

Introduction

Drug-induced liver injury (DILI) is one of the most frequent causes for drug withdrawal and the leading cause for compound attrition in drug development (Kullak-Ublick et al, 2017). The gold standard for toxicity testing, PHH, have a lack in bioavailability, high cost, high inter-donor variability and tendency to quickly dedif ferentiate upon plating. They can only provide limited mechanistic information (Lu et al, 2015). HepG2, a liver carcinoma cell line, displays many phenotypic features of normal liver cells, but has a low metabolic capacity compared to PHH. Current models that possess relatively high metabolic capacity such as HepaRG and Upcyte, show lower hepatotoxicity predictivity than HepG2 cells in comparison to PHH (Sison et al, 2017)

Objectives

Methods

Results

Conclusion