Scherf Jacob M, Hu Xiaoyang Serene, Tepp William H, Ichtchenko Konstantin, Johnson Eric A, Pellett Sabine
Department of Bacteriology, University of Wisconsin-Madison, Madison, Wisconsin, United States of America.
Department of Pharmacology, New York University School of Medicine, New York, New York, United States of America.
PLoS One. 2014 Oct 22;9(10):e111238. doi: 10.1371/journal.pone.0111238. eCollection 2014.
Botulinum neurotoxin type A1 (BoNT/A1) is a potent protein toxin responsible for the potentially fatal human illness botulism. Notwithstanding, the long-lasting flaccid muscle paralysis caused by BoNT/A has led to its utility as a powerful and versatile bio-pharmaceutical. The flaccid paralysis is due to specific cleavage of neuronal SNAREs by BoNTs. However, actions of BoNTs on intoxicated neurons besides the cleavage of SNAREs have not been studied in detail. In this study we investigated by microarray analysis the effects of BoNT/A and a catalytically inactive derivative (BoNT/A ad) on the transcriptome of human induced pluripotent stem cell (hiPSC)-derived neurons at 2 days and 2 weeks after exposure. While there were only minor changes in expression levels at 2 days post exposure, at 2 weeks post exposure 492 genes were differentially expressed more than 2-fold in BoNT/A1-exposed cells when compared to non-exposed populations, and 682 genes were differentially expressed in BoNT/A ad-exposed cells. The vast majority of genes were similarly regulated in BoNT/A1 and BoNT/A ad-exposed neurons, and the few genes differentially regulated between BoNT/A1 and BoNT/A ad-exposed neurons were differentially expressed less than 3.5 fold. These data indicate a similar response of neurons to BoNT/A1 and BoNT/A ad exposure. The most highly regulated genes in cells exposed to either BoNT/A1 or BoNT/A ad are involved in neurite outgrowth and calcium channel sensitization.
A型肉毒杆菌神经毒素1（BoNT/A1）是一种强效蛋白质毒素，可导致人类患上具有潜在致命性的肉毒中毒。尽管如此，BoNT/A所引起的持久弛缓性肌肉麻痹使其成为一种强大且用途广泛的生物制药。弛缓性麻痹是由于BoNTs对神经元SNAREs的特异性切割所致。然而，除了SNAREs的切割之外，BoNTs对中毒神经元的作用尚未得到详细研究。在本研究中，我们通过微阵列分析研究了BoNT/A和一种催化失活衍生物（BoNT/A ad）在暴露后2天和2周时对人诱导多能干细胞（hiPSC）衍生神经元转录组的影响。暴露后2天时表达水平仅有微小变化，而暴露后2周时，与未暴露群体相比，BoNT/A1暴露细胞中有492个基因差异表达超过2倍，BoNT/A ad暴露细胞中有682个基因差异表达。绝大多数基因在BoNT/A1和BoNT/A ad暴露的神经元中受到类似调控，BoNT/A1和BoNT/A ad暴露的神经元之间少数差异调控的基因差异表达小于3.5倍。这些数据表明神经元对BoNT/A1和BoNT/A ad暴露有类似反应。暴露于BoNT/A1或BoNT/A ad的细胞中调控程度最高的基因参与神经突生长和钙通道敏化。
