Human IPSC 3D brain model as a tool to study chemical-induced dopaminergic neuronal toxicity

David Pamies; Daphne Wiersma; Moriah E Katt; Liang Zhao; Johannes Burtscher; Georgina Harris; Lena Smirnova; Peter C Searson; Thomas Hartung; Helena T Hogberg

doi:10.1016/j.nbd.2022.105719

Human IPSC 3D brain model as a tool to study chemical-induced dopaminergic neuronal toxicity

Neurobiol Dis. 2022 Jul:169:105719. doi: 10.1016/j.nbd.2022.105719. Epub 2022 Apr 7.

Authors

Affiliations

¹ Center for Alternative to Animal Testing, Johns Hopkins University, 615 North Wolfe St., Baltimore, MD 21205, United States of America; Department of Biomedical Sciences, University of Lausanne, CH-1015 Lausanne, Switzerland. Electronic address: david.pamies@unil.ch.
² Center for Alternative to Animal Testing, Johns Hopkins University, 615 North Wolfe St., Baltimore, MD 21205, United States of America.
³ Department of Materials Science and Engineering, Johns Hopkins University, 3400 North Charles Street, Baltimore, MD 21218, United States of America; Institute of Nanobiotechnology, 100 Croft Hall, Johns Hopkins University, 3400 North Charles Street, Baltimore, MD 21218, United States of America.
⁴ Center for Alternative to Animal Testing, Johns Hopkins University, 615 North Wolfe St., Baltimore, MD 21205, United States of America; Bloomberg-Kimmel Institute for Cancer Immunotherapy, Johns Hopkins School of Medicine, Baltimore, MD 21287, United States of America.
⁵ Institute of Sport Sciences, University of Lausanne, CH-1015 Lausanne, Switzerland; Department of Biomedical Sciences, University of Lausanne, CH-1015 Lausanne, Switzerland.
⁶ Center for Alternative to Animal Testing, Johns Hopkins University, 615 North Wolfe St., Baltimore, MD 21205, United States of America; CAAT-Europe, University of Konstanz, Universitätstsr. 10, 78464 Konstanz, Germany.

Abstract

Oxidative stress is caused by an imbalance between the generation and detoxification of reactive oxygen and nitrogen species (ROS/RNS). This imbalance plays an important role in brain aging and age-related neurodegenerative diseases. In the context of Parkinson's disease (PD), the sensitivity of dopaminergic neurons in the substantia nigra pars compacta to oxidative stress is considered a key factor of PD pathogenesis. Here we study the effect of different oxidative stress-inducing compounds (6-OHDA, MPTP or MPP⁺) on the population of dopaminergic neurons in an iPSC-derived human brain 3D model (aka BrainSpheres). Treatment with 6-OHDA, MPTP or MPP⁺ at 4 weeks of differentiation disrupted the dopaminergic neuronal phenotype in BrainSpheres at (50, 5000, 1000 μM respectively). 6-OHDA increased ROS production and decreased mitochondrial function most efficiently. It further induced the greatest changes in gene expression and metabolites related to oxidative stress and mitochondrial dysfunction. Co-culturing BrainSpheres with an endothelial barrier using a transwell system allowed the assessment of differential penetration capacities of the tested compounds and the damage they caused in the dopaminergic neurons within the BrainSpheres In conclusion, treatment with compounds known to induce PD-like phenotypes in vivo caused molecular deficits and loss of dopaminergic neurons in the BrainSphere model. This approach therefore recapitulates common animal models of neurodegenerative processes in PD at similarly high doses. The relevance as tool for drug discovery is discussed.

Keywords: 3d culture; 6OHDA; MPP(+); MPTP; Microphysiological system; Organoid; Stem cells; Toxicant-induced Parkinson's disease; iPSC.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine / pharmacology
Animals
Brain / metabolism
Disease Models, Animal
Dopaminergic Neurons / metabolism
Humans
Induced Pluripotent Stem Cells* / metabolism
Mice
Mice, Inbred C57BL
Oxidopamine / pharmacology
Parkinson Disease* / metabolism
Reactive Oxygen Species / metabolism
Substantia Nigra / metabolism

Substances

Reactive Oxygen Species
Oxidopamine
1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine

Grants and funding

U18 TR000547/TR/NCATS NIH HHS/United States