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Src Family Kinase Inhibitors Antagonize the Toxicity of Multiple Serotypes of Botulinum Neurotoxin in Human Embryonic Stem Cell-Derived Motor Neurons

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Abstract

Botulinum neurotoxins (BoNTs), the causative agents of botulism, are potent inhibitors of neurotransmitter release from motor neurons. There are currently no drugs to treat BoNT intoxication after the onset of the disease symptoms. In this study, we explored how modulation of key host pathways affects the process of BoNT intoxication in human motor neurons, focusing on Src family kinase (SFK) signaling. Motor neurons derived from human embryonic stem (hES) cells were treated with a panel of SFK inhibitors and intoxicated with BoNT serotypes A, B, or E (which are responsible for >95 % of human botulism cases). Subsequently, it was found that bosutinib, dasatinib, KX2-391, PP1, PP2, Src inhibitor-1, and SU6656 significantly antagonized all three of the serotypes. Furthermore, the data indicated that the treatment of hES-derived motor neurons with multiple SFK inhibitors increased the antagonistic effect synergistically. Mechanistically, the small molecules appear to inhibit BoNTs by targeting host pathways necessary for intoxication and not by directly inhibiting the toxins’ proteolytic activity. Importantly, the identified inhibitors are all well-studied with some in clinical trials while others are FDA-approved drugs. Overall, this study emphasizes the importance of targeting host neuronal pathways, rather than the toxin’s enzymatic components, to antagonize multiple BoNT serotypes in motor neurons.

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Acknowledgments

We are indebted to Drs. Esta Sterneck and Balamurugan Kuppusamy for their insightful discussion concerning SFK inhibitors. Also, we thank Rajarshi Guha for R functions to optimize β for synergy calculations, and Veronica Soloveva for helpful discussion. This research was supported by grants from the Defense Threat Reduction Agency and National Institutes of Health (4R33AI101387 - 03). For JCB, this project has been funded in whole or in part with federal funds from the National Cancer Institute (NCI), National Institutes of Health (NIH), under contract no. HHSN261200800001E. LT has been supported by the Intramural Research Program of the NCI, Center for Cancer Research, NIH.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Geneva Foundation, Tacoma, WA, USA

    Erkan Kiris

  2. Department of Molecular and Translational Sciences, US Army Medical Research Institute of Infectious Diseases, 1425 Porter Street, Fort Detrick, Frederick, MD, 21702, USA

    Erkan Kiris, Jonathan E. Nuss, Laura M. Wanner, Glenn Y. Gomba, Krishna P. Kota, Rekha G. Panchal, Christopher D. Kane & Sina Bavari

  3. Neural Development Section, Mouse Cancer Genetics Program, Center for Cancer Research, NCI, Frederick, MD, 21702, USA

    Erkan Kiris, Hao T. Du & Lino Tessarollo

  4. Leidos Biomedical Research, Inc., Computational Drug Development Group (CDDG), NCI, Frederick, MD, 21702, USA

    James C. Burnett

  5. CDDG, Developmental Therapeutics Program, NCI, Frederick, MD, 21702, USA

    James C. Burnett, Brian D. Peyser & Rick Gussio

  6. Henry M. Jackson Foundation, Bethesda, MD, USA

    Christopher D. Kane

  7. DoD Biotechnology High Performance Computing Software Applications Institute (BHSAI), Telemedicine and Advanced Technology Research Center (TATRC), US Army Medical Research and Materiel Command (USAMRMC), Frederick, MD, 2170, USA

    Christopher D. Kane

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  1. Erkan Kiris
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  2. James C. Burnett
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  3. Jonathan E. Nuss
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Correspondence to Erkan Kiris or Sina Bavari.

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Kiris, E., Burnett, J.C., Nuss, J.E. et al. Src Family Kinase Inhibitors Antagonize the Toxicity of Multiple Serotypes of Botulinum Neurotoxin in Human Embryonic Stem Cell-Derived Motor Neurons. Neurotox Res 27, 384–398 (2015). https://doi.org/10.1007/s12640-015-9526-z

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