Abstract
Drug-induced liver injury (DILI) is one of the most common and serious adverse drug reactions and a major cause of drug development failure and withdrawal. Although different molecular mechanisms are implicated in DILI, enhanced ROS levels have been described as a major mechanism. Human-derived cell models are increasingly used in preclinical safety assessment because they provide quick and relatively inexpensive information in early stages of drug development. We have analyzed and compared the phenotype and functionality of two liver cell models (Upcyte human hepatocytes and HepaRG cells) to demonstrate their suitability for long-term hepatotoxicity assessments and mechanistic studies. The transcriptomic and functional analysis revealed the maintenance of phase I and phase II enzymes, and antioxidant enzymes along time in culture, although the differences found between both test systems underlie the differential sensitivity to hepatotoxins. The evaluation of several mechanisms of cell toxicity, including oxidative stress, by high-content screening, demonstrated that, by combining the stable phenotype of liver cells and repeated-dose exposure regimes to 12 test compounds at clinically relevant concentrations, both Upcyte hepatocytes and HepaRG offer suitable properties to be used in routine screening assays for toxicological assessments during drug preclinical testing.
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Abbreviations
- C max :
-
Therapeutic peak plasmatic concentration
- CYP:
-
Cytochrome P450
- DILI:
-
Drug-induced liver injury
- Fluo-4:
-
Fluo-4 acetoxymethyl ester
- GPX:
-
Glutathione peroxidase
- GSH:
-
Reduced glutathione
- GSR:
-
Glutathione reductase
- GST:
-
Glutathione S-reductase
- HCS:
-
High-content screening
- HLC:
-
Hepatocyte-like cells
- MEC:
-
Minimal effective concentration
- MMP:
-
Mitochondrial membrane potential
- PBGD:
-
Porphobilinogen deaminase
- PHH:
-
Primary human hepatocytes
- PI:
-
Propidium iodide
- TMRM:
-
Tetramethyl rhodamine methyl ester
- UGT:
-
UDP-glucuronosyltransferase
- UHH:
-
Upcyte human hepatocytes
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Acknowledgements
The scientific support from the Cytomics Unit from the Instituto de Investigación Sanitaria La Fe and the technical advice of Astrid Nörenberg is acknowledged.
Funding
This work has been supported by the Institute of Health Carlos III (ISCIII, Plan Estatal de I + D + i 2013–2016) and cofinanced by the European Regional Development Fund "A way to achieve Europe" (FEDER) through grants PI18/00993 and CP16/00097, by the Spanish Ministry of Science and Innovation Ministry-Spanish Research Agency through the Project PID2019-106000RB-C22/AEI/1 0.13039/501100011033, and by the Generalitat Valenciana (PROMETEO/2019/060). M.P. was supported by ACIF/2018/0226 (Generalitat Valenciana) and L.T. by ISCIII through grant CP16/00097.
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Conceptualization, MTD, LT: methodology, NJ, MP: writing-original draft preparation, MTD, LT: supervision, MTD, LT: writing-review and editing, MTD, LT: funding acquisition, MTD, LT All authors have read and agreed to the published version of the manuscript.
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The study was conducted according to the guidelines of the Declaration of Helsinki, and approved by the Ethics Committee of Health Research Institute La Fe (IIS La Fe) (2016/0226, date of approval 06/06/2017). Informed consent was obtained from all subjects involved in the study.
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Donato, M.T., Jiménez, N., Pelechá, M. et al. Oxidative-stress and long-term hepatotoxicity: comparative study in Upcyte human hepatocytes and hepaRG cells. Arch Toxicol 96, 1021–1037 (2022). https://doi.org/10.1007/s00204-022-03236-y
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DOI: https://doi.org/10.1007/s00204-022-03236-y