Assessment of multiple hiPSC-derived models for botulinum neurotoxin testing

Nicoleau, Camille; Lamotte, Juliette Duchesne de; Boude, Emilie; Noirmain, Fanny; Krupp, Johannes

doi:10.1016/j.toxicon.2018.11.206

Cited by 4 publications

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“…We have recently highlighted the potential of three additional hiPSC-derived neuronal models, including a Motor Neuron model, to provide a sensitive platform for studying BoNT intoxication mechanisms and for BoNT potency determination ( Nicoleau et al, 2018a ; Nicoleau et al, 2018b ). The potential of hiPSC-derived Motor Neurons for BoNT testing has been confirmed more recently by two studies ( Pellett et al, 2019 ; Schenke et al, 2020 ).…”

Section: Introductionmentioning

confidence: 99%

hiPSC-Derived Neurons Provide a Robust and Physiologically Relevant In Vitro Platform to Test Botulinum Neurotoxins

et al. 2021

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Botulinum neurotoxins (BoNTs) are zinc metalloproteases that block neurotransmitter release at the neuromuscular junction (NMJ). Their high affinity for motor neurons combined with a high potency have made them extremely effective drugs for the treatment of a variety of neurological diseases as well as for aesthetic applications. Current in vitro assays used for testing and developing BoNT therapeutics include primary rodent cells and immortalized cell lines. Both models have limitations concerning accuracy and physiological relevance. In order to improve the translational value of preclinical data there is a clear need to use more accurate models such as human induced Pluripotent Stem Cells (hiPSC)-derived neuronal models. In this study we have assessed the potential of four different human iPSC-derived neuronal models including Motor Neurons for BoNT testing. We have characterized these models in detail and found that all models express all proteins needed for BoNT intoxication and showed that all four hiPSC-derived neuronal models are sensitive to both serotype A and E BoNT with Motor Neurons being the most sensitive. We showed that hiPSC-derived Motor Neurons expressed authentic markers after only 7 days of culture, are functional and able to form active synapses. When cultivated with myotubes, we demonstrated that they can innervate myotubes and induce contraction, generating an in vitro model of NMJ showing dose-responsive sensitivity BoNT intoxication. Together, these data demonstrate the promise of hiPSC-derived neurons, especially Motor Neurons, for pharmaceutical BoNT testing and development.

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Section: Introductionmentioning

confidence: 99%

hiPSC-Derived Neurons Provide a Robust and Physiologically Relevant In Vitro Platform to Test Botulinum Neurotoxins

et al. 2021

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“…Formation of functional NMJ between hiPSC-derived motor neurons (hMNs) and rodent skeletal muscle [ 23 ], or primary human skeletal muscle [ 24 – 26 ], or even hiPSC-derived skeletal muscle [ 27 , 28 ] has recently been described. In this context, several studies assessed BoNTs activity on hMNs using biochemical assays that confirmed cleavage of hMN’s synaptosomal-associated protein 25 KDa (SNAP25) by BoNTs [ 29 – 32 ]. Our group recently described a human motor endplate system where hMNs and immortalized human muscle cells were cocultured in the same well and showed that the system is sensitive to BoNT/A, evidencing reduced/blockade of muscle cells contractions exposed to BoNT [ 33 ].…”

Section: Introductionmentioning

confidence: 99%

Optogenetically controlled human functional motor endplate for testing botulinum neurotoxins

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Background The lack of physiologically relevant and predictive cell-based assays is one of the major obstacles for testing and developing botulinum neurotoxins (BoNTs) therapeutics. Human-induced pluripotent stem cells (hiPSCs)-derivatives now offer the opportunity to improve the relevance of cellular models and thus the translational value of preclinical data. Methods We investigated the potential of hiPSC-derived motor neurons (hMNs) optical stimulation combined with calcium imaging in cocultured muscle cells activity to investigate BoNT-sensitivity of an in vitro model of human muscle-nerve system. Results Functional muscle-nerve coculture system was developed using hMNs and human immortalized skeletal muscle cells. Our results demonstrated that hMNs can innervate myotubes and induce contractions and calcium transient in muscle cells, generating an in vitro human motor endplate showing dose-dependent sensitivity to BoNTs intoxication. The implementation of optogenetics combined with live calcium imaging allows to monitor the impact of BoNTs intoxication on synaptic transmission in human motor endplate model. Conclusions Altogether, our findings demonstrate the promise of optogenetically hiPSC-derived controlled muscle-nerve system for pharmaceutical BoNTs testing and development.

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“…Many studies confirmed that iPSCs-derived neurons (i.e., motor neurons, cortical neurons, dopaminergic neurons, and sensory neurons) express all the necessary receptors and substrates for BoNT intoxication, and are sensitive for detection of several BoNTs serotypes with different potencies, making iPSC-derivatives cells of choice for BoNT research [ 96 , 157 , 160 , 161 , 162 , 163 ]. Motor neurons derived from hiPSCs have proved to be highly sensitive to BoNT intoxication [ 157 , 160 , 161 ]. Neurons derived from hiPSCs exhibited appropriate morphology, electrical behaviors, transsynaptic signaling and network activity.…”

Section: Emerging Cell-based Assays Using Human Ipscs Derivativesmentioning

confidence: 99%

Emerging Opportunities in Human Pluripotent Stem-Cells Based Assays to Explore the Diversity of Botulinum Neurotoxins as Future Therapeutics

Lamotte

Perrier

Martinat

et al. 2021

IJMS

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Botulinum neurotoxins (BoNTs) are produced by Clostridium botulinum and are responsible for botulism, a fatal disorder of the nervous system mostly induced by food poisoning. Despite being one of the most potent families of poisonous substances, BoNTs are used for both aesthetic and therapeutic indications from cosmetic reduction of wrinkles to treatment of movement disorders. The increasing understanding of the biology of BoNTs and the availability of distinct toxin serotypes and subtypes offer the prospect of expanding the range of indications for these toxins. Engineering of BoNTs is considered to provide a new avenue for improving safety and clinical benefit from these neurotoxins. Robust, high-throughput, and cost-effective assays for BoNTs activity, yet highly relevant to the human physiology, have become indispensable for a successful translation of engineered BoNTs to the clinic. This review presents an emerging family of cell-based assays that take advantage of newly developed human pluripotent stem cells and neuronal function analyses technologies.

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Assessment of multiple hiPSC-derived models for botulinum neurotoxin testing

Cited by 4 publications

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hiPSC-Derived Neurons Provide a Robust and Physiologically Relevant In Vitro Platform to Test Botulinum Neurotoxins

hiPSC-Derived Neurons Provide a Robust and Physiologically Relevant In Vitro Platform to Test Botulinum Neurotoxins

Optogenetically controlled human functional motor endplate for testing botulinum neurotoxins

Emerging Opportunities in Human Pluripotent Stem-Cells Based Assays to Explore the Diversity of Botulinum Neurotoxins as Future Therapeutics

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