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Cellular electrophysiological effects of botulinum toxin A on neonatal rat cardiomyocytes and on cardiomyocytes derived from human-induced pluripotent stem cells

Naunyn-Schmiedeberg's Archives of Pharmacology volume 396, pages 513–524 (2023)Cite this article

Abstract 

Botulinum toxin A is a well-known neurotransmitter inhibitor with a wide range of applications in modern medicine. Recently, botulinum toxin A preparations have been used in clinical trials to suppress cardiac arrhythmias, especially in the postoperative period. Its antiarrhythmic action is associated with inhibition of the nervous system of the heart, but its direct effect on heart tissue remains unclear. Accordingly, we investigate the effect of botulinum toxin A on isolated cardiac cells and on layers of cardiac cells capable of conducting excitation. Cardiomyocytes of neonatal rat pups and human cardiomyocytes obtained through cell reprogramming were used. A patch-clamp study showed that botulinum toxin A inhibited fast sodium currents and L-type calcium currents in a dose-dependent manner, with no apparent effect on potassium currents. Optical mapping showed that in the presence of botulinum toxin A, the propagation of the excitation wave in the layer of cardiac cells slows down sharply, conduction at high concentrations becomes chaotic, but reentry waves do not form. The combination of botulinum toxin A with a preparation of chitosan showed a stronger inhibitory effect by an order of magnitude. Further, the inhibitory effect of botulinum toxin A is not permanent and disappears after 12 days of cell culture in a botulinum toxin A-free medium. The main conclusion of the work is that the antiarrhythmic effect of botulinum toxin A found in clinical studies is associated not only with depression of the nervous system but also with a direct effect on heart tissue. Moreover, the combination of botulinum toxin A and chitosan reduces the effective dose of botulinum toxin A.

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Data availability

The raw/processed data required to reproduce these findings could be found at: https://drive.google.com/drive/folders/1EmvSYCro63dlE20vXIjHQ-tHpsUJpF65

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Acknowledgements

This work was carried out within the state assignment of the Ministry of Health of the Russian Federation (theme # 121031300224-1). We thank Suren Zakian’s lab for providing the hiPSCs of a healthy donor. This work was supported by own funds of the Moscow Institute of Physics and Technology and M. F. Vladimirsky Moscow Regional Research Clinical Institute, Moscow, Russia. The work was supported by the strategic academic leadership program “Priority 2030” (Agreement 075-02-2021-1316 30.09.2021).

Funding

Ministry of Health of the Russian Federation (project 121031300224–1)

• D. Sergeevichev

M. F. Vladimirsky Moscow Regional Research Clinical Institute, Moscow, Russia (own fundings)

• Sh. Frolova

• S. Kovalenko

• M. Slotvitsky

• K. Agladze

Moscow Institute of Physics and Technology (own fundings, strategic academic leadership program “Priority 2030”)

• V. Tsvelaya

• A. Nizamieva

• A. Nikitina

The funders had no role in study design, data collection and interpretation, or the decision to submit work for publication.

Author information

Authors and Affiliations

  1. Laboratory of Biophysics of Excitable Systems, Moscow Institute of Physics and Technology, Dolgoprudny, Russia

    Aygul Nizamieva, Sheida Frolova, Anna Nikitina & Konstantin Agladze

  2. M. F. Vladimirsky Moscow Regional Research Clinical Institute, Moscow, Russia

    Sheida Frolova, Mihail Slotvitsky, Sandaara Kovalenko, Valeriya Tsvelaya & Konstantin Agladze

  3. Laboratory of Experimental and Cellular Medicine, Moscow Institute of Physics and Technology, Dolgoprudny, Russia

    Sheida Frolova, Mihail Slotvitsky, Sandaara Kovalenko, Valeriya Tsvelaya & Konstantin Agladze

  4. “E. Meshalkin National Medical Research Center” of the Ministry of Health of the Russian Federation, 15 Rechkunovskaya St, Novosibirsk, Russia

    David Sergeevichev

Authors
  1. Aygul Nizamieva
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  2. Sheida Frolova
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  3. Mihail Slotvitsky
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  4. Sandaara Kovalenko
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  5. Valeriya Tsvelaya
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  6. Anna Nikitina
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  7. David Sergeevichev
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  8. Konstantin Agladze
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Contributions

All authors contributed to the study conception and design. Sh. Frolova and S. Kovalenko performed all patch-clamp studies. V. Tsvelaya and A. Nikitina conducted studies on human cardiomyocytes derived from IPSCs. A. Nizamieva and M. Slotvitsky performed substance studies on neonatal rat cardiomyocytes, processed all data, and prepared graphs and figures. D. Sergeevichev and K. Agladze wrote the main manuscript text. All authors read and approved the final manuscript. The authors declare that all data were generated in-house and that no paper mill was used.

Corresponding author

Correspondence to Valeriya Tsvelaya.

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Ethical approval

This study was performed in line with the principles of the Declaration of Helsinki and the Guide for the Care and Use of Laboratory Animals, published by the United States National Institutes of Health (Publication No. 85–23, revised 1996), and was approved by the Moscow Institute of Physics and Technology Life Science Center Provisional Animal Care and Research Procedures Committee, Protocol #A2-2012–09-02.

The cell line m34Sk3 is provided by the “E. Meshalkin National Medical Research Center” of the Ministry of Health of the Russian Federation and handling approved by the Institute of Circulation Pathology Ethics Committee (#27, March 21, 2013). All experiments and procedures were performed in accordance with principles for human experimentation as defined in the 1964 Declaration of Helsinki and its later amendments and were approved by the Scientific Council of the MIPT Life Science Center.

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Informed consent was obtained from all individual participants included in the study.

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The authors affirm that human research participants provided informed consent for publication of all figures, tables, and data included in this article.

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The authors declare no competing interests.

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Nizamieva, A., Frolova, S., Slotvitsky, M. et al. Cellular electrophysiological effects of botulinum toxin A on neonatal rat cardiomyocytes and on cardiomyocytes derived from human-induced pluripotent stem cells. Naunyn-Schmiedeberg's Arch Pharmacol 396, 513–524 (2023). https://doi.org/10.1007/s00210-022-02332-1

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  • DOI: https://doi.org/10.1007/s00210-022-02332-1

Keywords

  • Botulinum toxin A
  • Cardiotoxicity
  • Antiarrhythmic effect
  • Cardiomyocytes