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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Amacher DE (2011) The mechanistic basis for the induction of hepatic steatosis by xenobiotics. Exp Opin Drug Metab Toxicol 7(8):949–965. doi:10.1517/17425255.2011.577740
Amacher DE, Martin BA (1997) Tetracycline-induced steatosis in primary canine hepatocyte cultures. Fundam Appl Toxicol 40:256–263. doi:10.1006/faat.1997.2389
Anderson N, Borlak J (2008) Molecular mechanisms and therapeutic targets in steatosis and steatohepatitis. Pharmacol Rev 60:311–357. doi:10.1124/pr.108.00001
Angulo P (2002) Nonalcoholic fatty liver disease. N Engl J Med 346(16):1221–1231. doi:10.1056/NEJMra011775
Antherieu S, Rogue A, Fromenty B, Guillouzo A, Robin MA (2011) Induction of vesicular steatosis by amiodarone and tetracycline is associated with up-regulation of lipogenic genes in HepaRG Cells. Hepatology 53:1895–1905. doi:10.1002/hep.24290
Baxter MA, Rowe C, Alder J, Harrison S, Hanley KP, Park BK, Kitteringham NR, Goldring CE, Hanley NA (2010) Generating hepatic cell lineages from pluripotent stem cells for drug toxicity screening. Stem Cell Res 5:4–22. doi:10.1016/j.scr.2010.02.002
Brunt EM, Janney CG, Di Bisceglie AM, Neuschwander-Tetri BA, Bacon BR (1999) Nonalcoholic steatohepatitis: a proposal for grading and staging the histological lesions. Am J Gastroenterol 94:2467–2474. doi:10.1016/s0002-9270(99)00433-5
Charlton MR, Burns JM, Pedersen RA, Watt KD, Heimbach JK, Dierkhising RA (2011) Frequency and outcomes of liver transplantation for nonalcoholic steatohepatitis in the United States. Gastroenterology 141:1249–1253. doi:10.1053/j.gastro.2011.06.061
Davila JC, Cezar GG, Thiede M, Strom S, Miki T, Trosko J (2004) Use and application of stem cells in toxicology. Toxicol Sci 79:214–223. doi:10.1093/toxsci/kfh100
Donato MT, Jose Gomez-Lechon M (2012) Drug-induced liver steatosis and phospholipidosis: cell-based assays for early screening of drug candidates. Curr Drug Metab 13:1160–1173. doi:10.2174/138920012802850001
Donato MT, Martinez-Romero A, Jimenez N, Negro A, Herrera G, Castell JV, O’Connor J-E, Jose Gomez-Lechon M (2009) Cytometric analysis for drug-induced steatosis in HepG2 cells. Chem Biol Interact 181:417–423. doi:10.1016/j.cbi.2009.07.019
El-Serag HB, Rudolph L (2007) Hepatocellular carcinoma: epidemiology and molecular carcinogenesis. Gastroenterology 132:2557–2576. doi:10.1053/j.gastro.2007.04.061
Guguen-Guillouzo C, Guillouzo A (2010) General review on in vitro hepatocyte models and their applications. Methods Mol Biol 610:1–40. doi:10.1007/978-1-60761-688-7_1
Harrison SA, Torgerson S, Hayashi PH (2003) The natural history of nonalcoholic fatty liver disease: a clinical histopathological study. Am J Gastroenterol 98:2042–2047. doi:10.1016/s0002-9270(03)00622-1
Irizarry RA, Bolstad BM, Collin F, Cope LM, Hobbs B, Speed TP (2003) Summaries of affymetrix GeneChip probe level data. Nucleic Acids Res. doi:10.1093/nar/gng015
Kacew S, Lee BM (2013) Lu’s basic toxicology: fundamentals, target organs, and risk assessment, 6th edition Sam Kacew, Byung-Mu Lee. edition London: Informa Healthcare, pp 170–179
Kleiner DE, Gaffey MJ, Sallie R, Tsokos M, Nichols L, McKenzie R, Straus SE, Hoofnagle JH (1997) Histopathologic changes associated with fialuridine hepatotoxicity. Mod Pathol 10:192–199
Landesmann B, Goumenou M, Munn S, Whelan M (2012) Description of prototype modes-of-action related to repeated dose toxicity. JRC Sci Policy Report 75689
LeCluyse EL, Alexandre E (2010) Isolation and culture of primary hepatocytes from resected human liver tissue. Methods Mol Biol 640:57–82. doi:10.1007/978-1-60761-688-7_3
Matteoni CA, Younossi ZM, Gramlich T, Boparai N, Liu YC, McCullough AJ (1999) Nonalcoholic fatty liver disease: a spectrum of clinical and pathological severity. Gastroenterology 116:1413–1419. doi:10.1016/S0016-5085(99)70506-8
Moylan CA, Pang H, Dellinger A, Suzuki A, Garrett ME, Guy CD, Murphy SK, Ashley-Koch AE, Choi SS, Michelotti GA, Hampton DD, Chen Y, Tillmann HL, Hauser MA, Abdelmalek MF, Diehl AM (2014) Hepatic gene expression profiles differentiate presymptomatic patients with mild versus severe nonalcoholic fatty liver disease. Hepatology 59:471–482. doi:10.1002/hep.26661
Murphy DJ (2001) The biogenesis and functions of lipid bodies in animals, plants and microorganisms. Prog Lipid Res 40:325–438. doi:10.1016/s0163-7827(01)00013-3
Pagadala MR, McCullough AJ (2012) The relevance of liver histology to predicting clinically meaningful outcomes in nonalcoholic steatohepatitis. Clin Liver Dis 16:487–504. doi:10.1016/j.cld.2012.05.006
Rodrigues RM, De Kock J (2014) Human stem cell-derived hepatocytes: breakthrough of an expedient tool for preclinical assessment of drug-induced liver injury? Arch Toxicol 88:183–184. doi:10.1007/s00204-014-1199-5
Rodrigues RM, De Kock J, Branson S, Vinken M, Meganathan K, Chaudhari U, Sachinidis A, Govaere O, Roskams T, De Boe V, Vanhaecke T, Rogiers V (2014) Human skin-derived stem cells as a novel cell source for in vitro hepatotoxicity screening of pharmaceuticals. Stem Cells Dev 23:44–55. doi:10.1089/scd.2013.0157
Schuster D, Laggner C, Langer T (2005) Why drugs fail—a study on side effects in new chemical entities. Curr Pharm Des 11:3545–3559
Scott DA, Gholson CF, Netchvolodoff CV, Ray M, Gonzalez E, Bacon BR (1991) Incidental microvesicular steatosis due to valproic acid anticonvulsant therapy. Am J Gastroenterol 86:500–502
Snykers S, De Kock J, Rogiers V, Vanhaecke T (2009) In vitro differentiation of embryonic and adult stem cells into hepatocytes: state of the art. Stem Cells 27:577–605. doi:10.1634/stemcells.2008-0963
Szkolnicka D, Farnworth SL, Lucendo-Villarin B, Storck C, Zhou WL, Iredale JP, Flint O, Hay DC (2014) Accurate prediction of drug-induced liver injury using stem cell-derived populations. Stem Cells Transl Med 3:141–148. doi:10.5966/sctm.2013-0146
Uehara T, Ono A, Maruyama T, Kato I, Yamada H, Ohno Y, Urushidani T (2010) The Japanese toxicogenomics project: application of toxicogenomics. Mol Nutr Food Res 54:218–227. doi:10.1002/mnfr.200900169
Uetrecht J, Pessayre D, Mansouri A, Berson A, Fromenty B (2010) Mitochondrial involvement in drug-induced liver injury. Adverse Drug Reactions. Springer, Berlin, pp 311–365
Urushidani T, Nagao T (2005) Toxicogenomics: Japanese initiative. Handbook of toxicogenomics. Wiley-VCH Verlag GmbH & Co. KGaA, pp 623–631. doi:10.1002/3527603719.ch26
Vinken M (2013) The adverse outcome pathway concept: a pragmatic tool in toxicology. Toxicology 312:158–165. doi:10.1016/j.tox.2013.08.011
Wieckowska A, Feldstein AE (2008) Diagnosis of nonalcoholic fatty liver disease: invasive versus noninvasive. Semin Liver Dis 28:386–395. doi:10.1055/s-0028-1091983
Wobser H, Dorn C, Weiss TS, Amann T, Bollheimer C, Buttner R, Scholmerich J, Hellerbrand C (2009) Lipid accumulation in hepatocytes induces fibrogenic activation of hepatic stellate cells. Cell Res 19:996–1005. doi:10.1038/cr.2009.73
Youssef WI, McCullough AJ (2002) Steatohepatitis in obese individuals. Best Pract Res Clin Gastroenterol 16:733–747. doi:10.1053/bega.2002.0334
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.
Joery De Kock and Tamara Vanhaecke are equally contributing senior authors.
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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
- Postnatal stem cells
- In vitro toxicology