Uncovering the Functional Link Between SHANK3 Deletions and Deficiency in Neurodevelopment Using iPSC-Derived Human Neurons
Uncovering the Functional Link Between SHANK3 Deletions and Deficiency in Neurodevelopment Using iPSC-Derived Human Neurons
复制标题
使用 iPSC 衍生的人类神经元揭示 SHANK3 缺失与神经发育缺陷之间的功能联系
DOI：
10.3389/fnana.2019.00023
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发表时间：
2019-03-13
影响因子：
2.9
通讯作者：
Shi, Lingling
中科院分区：
文献类型：
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作者：
Huang, Guanqun;Chen, Shuting;Shi, Lingling
SHANK3突变，包括从头缺失，与自闭症谱系障碍（ASD）有关。然而，SHANK3功能丧失对神经发育的影响仍然知之甚少。在这里，我们在体外产生了人诱导多能干细胞（iPSC），然后进行神经分化和慢病毒介导的shRNA表达，以评估SHANK3敲低如何在多个时间点（长达4周）影响体外神经发育过程。我们发现，SHANK3敲低损害神经元发育的早期阶段和成熟神经元功能，如通过神经元索马大小、生长锥面积、神经突长度和分支数目的减少所证明的。值得注意的是，电生理学分析显示兴奋性和抑制性突触传递的缺陷。此外，转录组分析显示，在SHANK3敲低的细胞中，与神经元投射、运动和神经发生调节相关的多种生物学途径被破坏。总之，利用基于人类iPSC的神经诱导模型，本研究提出了形态学、电生理学和转录证据相结合的证据，支持SHANK3作为一种内在的细胞自主因子，控制人类神经元中的细胞功能发育。
SHANK3 mutations, including de novo deletions, have been associated with autism spectrum disorders (ASD). However, the effects of SHANK3 loss of function on neurodevelopment remain poorly understood. Here we generated human induced pluripotent stem cells (iPSC) in vitro, followed by neuro-differentiation and lentivirus-mediated shRNA expression to evaluate how SHANK3 knockdown affects the in vitro neurodevelopmental process at multiple time points (up to 4 weeks). We found that SHANK3 knockdown impaired both early stage of neuronal development and mature neuronal function, as demonstrated by a reduction in neuronal soma size, growth cone area, neurite length and branch numbers. Notably, electrophysiology analyses showed defects in excitatory and inhibitory synaptic transmission. Furthermore, transcriptome analyses revealed that multiple biological pathways related to neuron projection, motility and regulation of neurogenesis were disrupted in cells with SHANK3 knockdown. In conclusion, utilizing a human iPSC-based neural induction model, this study presented combined morphological, electrophysiological and transcription evidence that support that SHANK3 as an intrinsic, cell autonomous factor that controls cellular function development in human neurons.