In vitro model for drug-induced phospholipidosis

Commonly used acronym: drug-induced phospholipidosis (DIPL)

Scope of the method

Alternative method relates to
  • Human health
Alternative method is situated in
  • Basic Research
  • Translational - Applied Research
Type of alternative method
  • In vitro - Ex vivo
This method makes use of
  • Human derived cells / tissues / organs
Specify the type of cells/tissues/organs
liver

Description

Method keywords
  • Human skin stem cells-derived hepatic cells
  • liver embryogenesisin vitro
  • drug screening
  • drug-induced liver injury
Scientific area keywords
  • liver
Method desription

Human skin stem cells-derived hepatic cells (hSKP-HPC) have already shown to be a good human in vitro tool to screen for compounds that can induce hepatic steatosis and acute liver failure. Another important liver disorder that is often reason of discontinuation of chemicals in drug discovery is drug-induced phospholipidosis. Phospholipidosis is a metabolic disorder characterized by an excessive intracellular accumulation of phospholipids. The objective of my research is the application of hSKP-HPC to investigate liver phospholipidosis and the elucidation of the mechanisms leading to this pathological condition in vitro. Hereby, hSKP-HPC are cultured in a standard two dimensional (2D) setup. Data show that hSKP-HPC exposed to amiodarone, which is known to induce PL in vivo, accumulate phospholipids and neutral lipids intracellularly. Typical lamellar inclusions can also be identified by transmission electron microscopy. Significant upregulation of genes involved in lysosomal activity and biosynthesis of new phospholipids is found in our cell model.

Method status
  • Validated by an external party (e.g. OECD, EURL ECVAM,…)

Pros, cons & Future potential

Advantages

Strenghts are represented by inter-donor variability reflecting the human situation

Future & Other applications

Potential application of this cell model in related to drug toxicity screening platform

References, associated documents and other information

References

Rodrigues RM, Heymans A, De Boe V, Sachinidis A, Chaudhari U, Govaere O, Roskams T, Vanhaecke T, Rogiers V, De Kock J. ,Toxicol Lett. 2016 Jan 5;240(1):50-9. doi: 10.1016/j.toxlet.2015.10.014. Epub 2015 Oct 20 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., Stem Cells Dev. 2014 Jan 1;23(1):44-55. doi: 10.1089/scd.2013.0157. Epub 2013 Sep 21.

Associated documents
Other remarks

Experimental design: 1- Human skin stem cells are differentiated towards hepatic cells for 24 days 2- Differentiated cells are exposed to the drug of interest (amiodarone, in this case) 3- Quantification of lipids is performed using flow cytometry 4- Modulation of gene expression under drug exposure is evaluated by qPCR Controls: cells non-exposed to the drug represent the control group Data interpretation and acceptance criteria: stastisticel analysis Performance: yes Applicability domain: amiodarone( in my study) , valproic acid and acetaminophen were already tested in this cell model. I am not aware of compounds that cannot be used on the cell model since they were not tested yet.

Contact person

Mieke Van Mulders

Organisations

Sciensano
Chemical and physical health risks
Belgium

VUB
IVTD
Belgium