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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

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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

References

  • Adler M, Pellett S, Sharma SK, Lebeda FJ, Dembek ZF, Mahan MA (2022) Preclinical evidence for the role of botulinum neurotoxin A (BoNT/A) in the treatment of peripheral nerve injury. Microorganisms 10(5):886

    Article  CAS  Google Scholar 

  • Aranki SF, Shaw DP, Adams DH, Rizzo RJ, Couper GS, VanderVliet M, ..., Burstin HR (1996) Predictors of atrial fibrillation after coronary artery surgery: current trends and impact on hospital resources. Circulation 94(3):390-397

  • Atienza F, Jalife J (2007) Reentry and atrial fibrillation. Heart Rhythm 4(3):S13–S16

    Article  Google Scholar 

  • Buckley U, Rajendran PS, Shivkumar K (2017) Ganglionated plexus ablation for atrial fibrillation: just because we can, does that mean we should? Heart Rhythm 14(1):133–134

    Article  Google Scholar 

  • Burridge PW, Matsa E, Shukla P, Lin ZC, Churko JM, Ebert AD, ... Wu JC (2014) Chemically defined generation of human cardiomyocytes. Nat Methods 11(8):855-860

  • Cocco A, Albanese A (2018) Recent developments in clinical trials of botulinum neurotoxins. Toxicon 147:77–83

    Article  CAS  Google Scholar 

  • Courtemanche M, Ramirez RJ, Nattel S (1998) Ionic mechanisms underlying human atrial action potential properties: insights from a mathematical model. Am J Physiol Heart Circ Physiol 275(1):H301–H321

    Article  CAS  Google Scholar 

  • Deerenberg EB, Shao JM, Elhage SA, Lopez R, Ayuso SA, Augenstein VA, Heniford BT (2021) Preoperative botulinum toxin A injection in complex abdominal wall reconstruction–a propensity-scored matched study. Am J Surg 222(3):638–642

    Article  Google Scholar 

  • Frolova SR, Gaiko O, Tsvelaya VA, Pimenov OY, Agladze KI (2016) Photocontrol of voltage-gated ion channel activity by azobenzene trimethylammonium bromide in neonatal rat cardiomyocytes. PLoS ONE 11(3):e0152018

    Article  Google Scholar 

  • Honarbakhsh S, Schilling RJ, Orini M, Providencia R, Keating E, Finlay M, ... Hunter RJ (2019) Structural remodeling and conduction velocity dynamics in the human left atrium: relationship with reentrant mechanisms sustaining atrial fibrillation. Heart Rhythm 16(1):18-25

  • Kernik DC, Morotti S, Wu H, Garg P, Duff HJ, Kurokawa J, ... Clancy CE (2019) A computational model of induced pluripotent stem‐cell derived cardiomyocytes incorporating experimental variability from multiple data sources. J Physiol 597(17):4533-4564

  • Kimura K, Kimura H, Yokosawa N, Isogai H, Isogai E, Kozaki S, ... Fujii N (1998) Negative chronotropic effect of botulinum toxin A on neonatal rat cardiac myocytes. Biochem Biophys Res Commun 244(1):275-279

  • Kléber AG, Rudy Y (2004) Basic mechanisms of cardiac impulse propagation and associated arrhythmias. Physiol Rev 84(2):431–488

    Article  Google Scholar 

  • Lian X, Zhang J, Azarin SM, Zhu K, Hazeltine LB, Bao X, ... Palecek SP (2013) Directed cardiomyocyte differentiation from human pluripotent stem cells by modulating Wnt/β-catenin signaling under fully defined conditions. Nat Protoc 8(1):162-175

  • Lippiat JD (2008) Whole-cell recording using the perforated patch clamp technique. In Potassium Channels (pp. 141–149). Humana Press

  • Lo LW, Scherlag BJ, Chang HY, Lin YJ, Chen SA, Po SS (2013) Paradoxical long-term proarrhythmic effects after ablating the “head station” ganglionated plexi of the vagal innervation to the heart. Heart Rhythm 10(5):751–757

    Article  Google Scholar 

  • Lo LW, Chang HY, Scherlag BJ, Lin YJ, Chou YH, Lin WL, ... Po SS (2016) Temporary suppression of cardiac ganglionated plexi leads to long‐term suppression of atrial fibrillation: evidence of early autonomic intervention to break the vicious cycle of “AF begets AF”. J Am Heart Assoc 5(7):e003309

  • Meijer van Putten RM, Mengarelli I, Guan K, Zegers JG, van Ginneken AC, Verkerk AO, Wilders R (2015) Ion channelopathies in human induced pluripotent stem cell derived cardiomyocytes: a dynamic clamp study with virtual IK1. Front Physiol 6:7

    Article  Google Scholar 

  • Nazeri A, Ganapathy AV, Massumi A, Massumi M, Tuzun E, Stainback R, ... Razavi M (2017) Effect of botulinum toxin on inducibility and maintenance of atrial fibrillation in ovine myocardial tissue. Pacing Clin Electrophysiol 40(6):693-702

  • Nicolas J, Hendriksen PJ, de Haan LH, Koning R, Rietjens IM, Bovee TF (2015) In vitro detection of cardiotoxins or neurotoxins affecting ion channels or pumps using beating cardiomyocytes as alternative for animal testing. Toxicol in Vitro 29(2):281–288

    Article  CAS  Google Scholar 

  • Oh S, Choi EK, Choi YS (2010) Short-term autonomic denervation of the atria using botulinum toxin A. Korean Circ J 40(8):387–390

    Article  Google Scholar 

  • Oh S, Choi EK, Zhang, Y., & Mazgalev, T. N. (2011). Botulinum toxin A injection in epicardial autonomic ganglia temporarily suppresses vagally mediated atrial fibrillation. Circ Arrhythm Electrophysiol 4(4):560–565

  • O’Hara T, Virág L, Varró A, Rudy Y (2011) Simulation of the undiseased human cardiac ventricular action potential: model formulation and experimental validation. PLoS Comput Biol 7(5):e1002061

    Article  CAS  Google Scholar 

  • Piccini JP, Ahlsson A, Dorian P, Gillinov MA, Kowey PR, Mack MJ, ... Benussi S (2022) Design and rationale of a phase 2 study of neurotoxin (botulinum toxin type A) for the prevention of post-operative atrial fibrillation–the NOVA study. Am Heart J 245:51-59

  • Podgurskaya AD, Tsvelaya VA, Slotvitsky MM, Dementyeva EV, Valetdinova KR, Agladze KI (2019) The use of iPSC-derived cardiomyocytes and optical mapping for erythromycin arrhythmogenicity testing. Cardiovasc Toxicol 19(6):518–528

    Article  CAS  Google Scholar 

  • Pokushalov E, Kozlov B, Romanov A, Strelnikov A, Bayramova S, Sergeevichev D, ... Steinberg JS (2014) Botulinum toxin A injection in epicardial fat pads can prevent recurrences of atrial fibrillation after cardiac surgery: results of a randomized pilot study. J Am Coll Cardiol 64(6):628-629

  • Pokushalov E, Kozlov B, Romanov A, Strelnikov A, Bayramova S, Sergeevichev D, ... Steinberg JS (2015) Long-term suppression of atrial fibrillation by botulinum toxin A injection into epicardial fat pads in patients undergoing cardiac surgery: one-year follow-up of a randomized pilot study. Circ Arrhythm Electrophysiol 8(6):1334–1341

  • Romanov A, Pokushalov E, Ponomarev D, Bayramova S, Shabanov V, Losik D, ... Steinberg JS (2019) Long-term suppression of atrial fibrillation by botulinum toxin injection into epicardial fat pads in patients undergoing cardiac surgery: three-year follow-up of a randomized study. Heart Rhythm 16(2):172-177

  • Rostagno C, Blanzola C, Pinelli F, Rossi A, Carone E, Stefàno PL (2014) Atrial fibrillation after isolated coronary surgery. Incidence, long term effects and relation with operative technique. Heart, Lung and Vessels 6(3):171

  • Sergeevichev DS, Krasilnikova AA, Strelnikov AG, Fomenko VV, Salakhutdinov NF, Romanov AB, ... Steinberg JS (2018) Globular chitosan prolongs the effective duration time and decreases the acute toxicity of botulinum neurotoxin after intramuscular injection in rats. Toxicon 143:90-95

  • Sergeevichev D, Fomenko V, Strelnikov A, Dokuchaeva A, Vasilieva M, Chepeleva E, ... Chernyavskiy A (2020) Botulinum toxin-chitosan nanoparticles prevent arrhythmia in experimental rat models. Mar Drugs 18(8):410

  • Shin MC, Wakita M, Xie DJ, Yamaga T, Iwata S, Torii Y, ..., Akaike N (2011) Inhibition of membrane Na+ channels by A type botulinum toxin A at femtomolar concentrations in central and peripheral neurons. J Pharmacol Sci 1112090628–1112090628

  • Slotvitsky MM, Tsvelaya VA, Podgurskaya AD, Agladze KI (2020) Formation of an electrical coupling between differentiating cardiomyocytes. Sci Rep 10(1):1–11

    Article  Google Scholar 

  • Sung DJ, Kim JG, Won KJ, Kim B, Shin HC, Park JY, Bae YM (2012) Blockade of K+ and Ca2+ channels by azole antifungal agents in neonatal rat ventricular myocytes. Biol Pharm Bull 35(9):1469–1475

    Article  CAS  Google Scholar 

  • Tanyeli O, Isik M (2020) Botulinum toxin treatment in cardiovascular surgery. In Botulinum Toxin Treatment in Surgery, Dentistry, and Veterinary Medicine (pp. 157–171). Springer, Cham

  • Ten Tusscher KH, Panfilov AV (2006) Alternans and spiral breakup in a human ventricular tissue model. Am J Physiol Heart Circ Physiol 291(3):H1088–H1100

    Article  Google Scholar 

  • Tsuboi M, Furukawa Y, Kurogouchi F, Nakajima K, Hirose M, Chiba S (2002) Botulinum neurotoxin A blocks cholinergic ganglionic neurotransmission in the dog heart. Jpn J Pharmacol 89(3):249–254

    Article  CAS  Google Scholar 

  • Waldron NH, Cooter M, Haney JC, Schroder JN, Gaca JG, Lin SS, ... Mathew JP (2019) Temporary autonomic modulation with botulinum toxin type A to reduce atrial fibrillation after cardiac surgery. Heart Rhythm 16(2):178-184

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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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