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The Use of Human Induced Pluripotent Stem Cells for Testing Neuroprotective Activity of Pharmacological Compounds

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Abstract

Development of therapeutic preparations involves several steps, starting with the synthesis of chemical compounds and testing them in different models for selecting the most effective and safest ones to clinical trials and introduction into medical practice. Cultured animal cells (both primary and transformed) are commonly used as models for compound screening. However, cell models display a number of disadvantages, including insufficient standardization (primary cells) and disruption of cell genotypes (transformed cells). Generation of human induced pluripotent stem cells (IPSCs) offers new possibilities for the development of high-throughput test systems for screening potential therapeutic preparations with different activity spectra. Due to the capacity to differentiate into all cell types of an adult organism, IPSCs are a unique model that allows examining the activity and potential toxicity of tested compounds during the entire differentiation process in vitro. In this work, we demonstrated the efficiency of IPSCs and their neuronal derivatives for selecting substances with the neuroprotective activity using two classes of compounds — melanocortin family peptides and endocannabinoids. None of the tested compounds displayed cyto- or embryotoxicity. Both melanocortin peptides and endocannabinoids exerted neuroprotective effect in the neuronal precursors and IPSC-derived neurons subjected to hydrogen peroxide. The endo-cannabinoid N-docosahexaenoyl dopamine exhibited the highest neuroprotective effect (∼70%) in the differentiated cultures enriched with dopaminergic neurons; the effect of melanocortin Semax was ∼40%. The possibility of using other IPSC derivatives for selecting compounds with the neuroprotective activity is discussed.

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Abbreviations

alpha-MSH:

alpha-melanocyte-stimulating hormone

BDNF:

brain-derived neurotrophic factor

DA neuron:

dopaminergic neuron

DMSO:

dimethyl sulfoxide

EC:

endocannabinoid

G418:

geneticin

GDNF:

glial cell-derived neurotrophic factor

IPSC:

induced pluripotent stem cell

MC:

melanocortin

N-ADA:

N-arachidonoyl dopamine

N-DDA:

N-docosahexaenoyl dopamine

PGP:

melanocortin family peptide Pro-Gly-Pro

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Acknowledgements

The authors express gratitude to V. V. Bezuglov (Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences) for kindly providing endocannabinoids for this study.

Funding

This work was supported by the Russian Foundation for Basic Research (project 17-04-01661a) and the Program of the Presidium of the Russian Academy of Sciences “Fundamental studies for biomedical technologies”. The equipment used in the study was provided by the Center of Collective Use of the Institute of Molecular Genetics, Russian Academy of Sciences (Center for Cell and Gene Technologies).

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Authors and Affiliations

  1. Institute of Molecular Genetics, Russian Academy of Sciences, 123182, Moscow, Russia

    E. V. Novosadova, E. L. Arsenyeva, S. A. Antonov, Y. N. Vanyushina, T. V. Malova, A. A. Komissarov, L. A. Andreeva, N. F. Myasoedov, V. Z. Tarantul & I. A. Grivennikov

  2. Research Center of Neurology, 125367, Moscow, Russia

    S. N. Illarioshkin & L. G. Khaspekov

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  1. E. V. Novosadova
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  2. E. L. Arsenyeva
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  3. S. A. Antonov
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  4. Y. N. Vanyushina
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  5. T. V. Malova
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  6. A. A. Komissarov
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  7. S. N. Illarioshkin
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  8. L. G. Khaspekov
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  9. L. A. Andreeva
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  10. N. F. Myasoedov
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  11. V. Z. Tarantul
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  12. I. A. Grivennikov
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Correspondence to E. V. Novosadova or I. A. Grivennikov.

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The authors declare no conflict of interest in financial or any other sphere.

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This article does not contain any studies with human participants or animals performed by any of the authors.

Russian Text © The Author(s), 2019, published in Biokhimiya, 2019, Vol. 84, No. 11, pp. 1610–1621.

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Novosadova, E.V., Arsenyeva, E.L., Antonov, S.A. et al. The Use of Human Induced Pluripotent Stem Cells for Testing Neuroprotective Activity of Pharmacological Compounds. Biochemistry Moscow 84, 1296–1305 (2019). https://doi.org/10.1134/S0006297919110075

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