Embryonic stem cell-derived motoneurons provide a highly sensitive cell culture model for botulinum neurotoxin studies, with implications for high-throughput drug discovery.
Embryonic stem cell-derived motoneurons provide a highly sensitive cell culture model for botulinum neurotoxin studies, with implications for high-throughput drug discovery.
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DOI：
10.1016/j.scr.2011.01.002
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发表时间：
2011-05
影响因子：
1.2
通讯作者：
Bavari, Sina
中科院分区：
文献类型：
--
作者：
Kiris, Erkan;Nuss, Jonathan E.;Burnett, James C.;Kota, Krishna P.;Koh, Dawn C.;Wanner, Laura M.;Torres-Melendez, Edna;Gussio, Rick;Tessarollo, Lino;Bavari, Sina
肉毒神经毒素（BoNT）通过特异性切割对神经递质胞吐至关重要的蛋白质来抑制胆碱能突触传递。由于这些毒素的致命性，人们更加担心它们可能被用作生物恐怖主义制剂。此外，它们广泛用于美容目的和医疗，增加了意外过量和环境暴露的潜在风险。因此，迫切需要开发新的方式来对抗BoNT中毒。哺乳动物运动神经元是BoNT的主要靶点，然而，由于分离细胞所需的程序的困难和效率差，它们不适合高通量药物筛选测定。在这里，我们探讨了适合的胚胎干（ES）细胞衍生的运动神经元作为一个可再生的，可重复的，生理相关的系统BoNT的研究。我们发现，ES-衍生的运动神经元的敏感性BoNT/A中毒是相当的原代小鼠脊髓运动神经元。此外，我们证明了几种BoNT/A抑制剂保护SNAP-25，BoNT/A底物，在ES衍生的运动神经元系统。此外，该系统与基于免疫荧光的高通量研究兼容。这些数据表明，ES衍生的运动神经元提供了一个高度敏感的系统，适合大规模筛选，以快速识别和评估新疗法的生物学功效。
Botulinum neurotoxins (BoNTs) inhibit cholinergic synaptic transmission by specifically cleaving proteins that are crucial for neurotransmitter exocytosis. Due to the lethality of these toxins, there are elevated concerns regarding their possible use as bioterrorism agents. Moreover, their widespread use for cosmetic purposes, and as medical treatments, has increased the potential risk of accidental overdosing and environmental exposure. Hence, there is an urgent need to develop novel modalities to counter BoNT intoxication. Mammalian motoneurons are the main target of BoNTs, however, due to the difficulty and poor efficiency of the procedures required to isolate the cells, they are not suitable for high-throughput drug screening assays. Here, we explored the suitability of embryonic stem (ES) cell-derived motoneurons as a renewable, reproducible, and physiologically relevant system for BoNT studies. We found that the sensitivity of ES-derived motoneurons to BoNT/A intoxication is comparable to that of primary mouse spinal motoneurons. Additionally, we demonstrated that several BoNT/A inhibitors protected SNAP-25, the BoNT/A substrate, in the ES-derived motoneuron system. Furthermore, this system is compatible with immunofluorescence-based high-throughput studies. These data suggest that ES-derived motoneurons provide a highly sensitive system that is amenable to large-scale screenings to rapidly identify and evaluate the biological efficacies of novel therapeutics.
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影响因子：
56.9
作者：
Dong, M;Yeh, F;Chapman, ER
通讯作者：
Chapman, ER
DOI：
10.1073/pnas.0903111106
发表时间：
2009-06-09
影响因子：
11.1
作者：
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DOI：
10.1124/jpet.108.147744
发表时间：
2009-07-01
影响因子：
3.5
作者：
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通讯作者：
Yan, Xiuzhen
DOI：
10.1038/nrd1694
发表时间：
2005-04
期刊：
Nature reviews. Drug discovery
影响因子：
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作者：
Burnett JC;Henchal EA;Schmaljohn AL;Bavari S
通讯作者：
Bavari S
DOI：
10.1073/pnas.0401103101
发表时间：
2004-05-04
影响因子：
11.1
作者：
Harper, JM;Krishnan, C;Kerr, DA
通讯作者：
Kerr, DA