Integrating distribution kinetics and toxicodynamics to assess repeat dose neurotoxicity in vitro using human BrainSpheres: a case study on amiodarone

Carolina Nunes; Susana Proença; Giovanna Ambrosini; David Pamies; Aurélien Thomas; Nynke I Kramer; Marie-Gabrielle Zurich

doi:10.3389/fphar.2023.1248882

Integrating distribution kinetics and toxicodynamics to assess repeat dose neurotoxicity in vitro using human BrainSpheres: a case study on amiodarone

Front Pharmacol. 2023 Sep 6:14:1248882. doi: 10.3389/fphar.2023.1248882. eCollection 2023.

Authors

Carolina Nunes^{1

2}, Susana Proença^{3

4}, Giovanna Ambrosini^{5

6}, David Pamies^{1

2}, Aurélien Thomas^{7

8}, Nynke I Kramer^{3

4}, Marie-Gabrielle Zurich^{1

2}

Affiliations

¹ Department of Biomedical Sciences, University of Lausanne, Lausanne, Switzerland.
² Swiss Centre for Applied Human Toxicology (SCAHT), Basel, Switzerland.
³ Institute for Risk Assessment Sciences, Utrecht University, Utrecht, Netherlands.
⁴ Toxicology Division, Wageningen University, Wageningen, Netherlands.
⁵ Bioinformatics Competence Center, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
⁶ Bioinformatics Competence Center, University of Lausanne, Lausanne, Switzerland.
⁷ Unit of Forensic Toxicology and Chemistry, CURML, Lausanne and Geneva University Hospitals, Geneva, Switzerland.
⁸ Faculty Unit of Toxicology, CURML, Faculty of Biology and Medicine, University of Lausanne, Lausanne, Switzerland.

Abstract

For ethical, economical, and scientific reasons, animal experimentation, used to evaluate the potential neurotoxicity of chemicals before their release in the market, needs to be replaced by new approach methodologies. To illustrate the use of new approach methodologies, the human induced pluripotent stem cell-derived 3D model BrainSpheres was acutely (48 h) or repeatedly (7 days) exposed to amiodarone (0.625-15 µM), a lipophilic antiarrhythmic drug reported to have deleterious effects on the nervous system. Neurotoxicity was assessed using transcriptomics, the immunohistochemistry of cell type-specific markers, and real-time reverse transcription-polymerase chain reaction for various genes involved in the lipid metabolism. By integrating distribution kinetics modeling with neurotoxicity readouts, we show that the observed time- and concentration-dependent increase in the neurotoxic effects of amiodarone is driven by the cellular accumulation of amiodarone after repeated dosing. The development of a compartmental in vitro distribution kinetics model allowed us to predict the change in cell-associated concentrations in BrainSpheres with time and for different exposure scenarios. The results suggest that human cells are intrinsically more sensitive to amiodarone than rodent cells. Amiodarone-induced regulation of lipid metabolism genes was observed in brain cells for the first time. Astrocytes appeared to be the most sensitive human brain cell type in vitro. In conclusion, assessing readouts at different molecular levels after the repeat dosing of human induced pluripotent stem cell-derived BrainSpheres in combination with the compartmental modeling of in vitro kinetics provides a mechanistic means to assess neurotoxicity pathways and refine chemical safety assessment for humans.

Keywords: hiPSC; in silico modeling; in vitro distribution kinetics; lipid metabolism; microphysiological system; neurotoxicity; spheroid.

Grants and funding

This work was funded by the European Union’s Horizon 2020 Research and Innovation Program Marie Sklodowska-Curie Actions Innovative Training Network project in3, under grant agreement no. 721975 (fellowships to CN and SP).