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In vitro assessment of drug-induced liver steatosis based on human dermal stem cell-derived hepatic cells

Abstract

Steatosis, also known as fatty liver disease (FLD), is a disorder in which the lipid metabolism of the liver is disturbed, leading to the abnormal retention of lipids in hepatocytes. FLD can be induced by several drugs, and although it is mostly asymptomatic, it can lead to steatohepatitis, which is associated with liver inflammation and damage. Drug-induced liver injury is currently the major cause of postmarketing withdrawal of pharmaceuticals and discontinuation of the development of new chemical entities. Therefore, the potential induction of steatosis must be evaluated during preclinical drug development. However, robust human-relevant in vitro models are lacking. In the present study, we explore the applicability of hepatic cells (hSKP-HPCs) derived from postnatal skin precursors, a stem cell population residing in human dermis, to investigate the steatosis-inducing effects of sodium valproate (Na-VPA). Exposure of hSKP-HPC to sub-cytotoxic concentrations of this reference steatogenic compound showed an increased intracellular accumulation of lipid droplets, and the modulation of key factors involved in lipid metabolism. Using a toxicogenomics approach, we further compared Na-VPA-treated hSKP-HPC and Na-VPA-treated primary human hepatocytes to liver samples from patients suffering from mild and advanced steatosis. Our data show that in hSKP-HPC exposed to Na-VPA and liver samples of patients suffering from mild steatosis, but not in primary human hepatocytes, “liver steatosis” was efficiently identified as a toxicological response. These findings illustrate the potential of hSKP-HPC as a human-relevant in vitro model to identify hepatosteatotic effects of chemical compounds.

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Acknowledgments

The authors would like to thank Mrs. Marleen Berghmans for her technical support in the processing of samples and acquisition of TEM images. This work received funding from the Fund for Scientific Research in Flanders (FWO), the European Commission’s FP7 under grant agreement number 266838 (DETECTIVE), and from ISRIB (Brustem) and BELSPO (IAP-HEPRO).

Conflict of interest

There are no competing financial interests.

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Correspondence to Robim M. Rodrigues.

Additional information

Joery De Kock and Tamara Vanhaecke are equally contributing senior authors.

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Figure 1

Supplementary material LipidTOX Green neutral lipid stain of hHEP exposed to NA-VPA (A) (5 mM, 72 h) and non-exposed controls (B). (TIFF 8003 kb)

Supplementary material 2 (TIFF 170 kb)

Supplementary material 3 (DOCX 22 kb)

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Rodrigues, R.M., Branson, S., De Boe, V. et al. In vitro assessment of drug-induced liver steatosis based on human dermal stem cell-derived hepatic cells. Arch Toxicol 90, 677–689 (2016). https://doi.org/10.1007/s00204-015-1483-z

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Keywords

  • Postnatal stem cells
  • hSKP
  • Steatosis
  • In vitro toxicology