Deciphering the Proteome Dynamics during Development of Neurons Derived from Induced Pluripotent Stem Cells

Neuronal development is a complex multistep process that shapes neurons by progressing though several typical stages, including axon outgrowth, dendrite formation, and synaptogenesis. Knowledge of the mechanisms of neuronal development is mostly derived from the study of animal models. Advances in stem cell technology now enable us to generate neurons from human induced pluripotent stem cells (iPSCs). Here we provide a mass spectrometry-based quantitative proteomic signature of human iPSC-derived neurons, i.e., iPSC-derived induced glutamatergic neurons and iPSC-derived motor neurons, throughout neuronal differentiation. Tandem mass tag 10-plex labeling was carried out to perform proteomic profiling of cells at different time points. Our analysis reveals significant expression changes (FDR < 0.001) of several key proteins during the differentiation process, e.g., proteins involved in the Wnt and Notch signaling pathways. Overall, our data provide a rich resource of information on protein expression during human iPSC neuron differentiation.

神经元发育是一类复杂的多阶段过程，通过历经多个典型阶段塑造神经元的形态与功能，典型阶段涵盖轴突生长、树突形成与突触发生。目前学界对神经元发育机制的认知大多来自动物模型研究。如今干细胞技术的发展已可让我们从人类诱导多能干细胞（induced pluripotent stem cells，iPSCs）中诱导生成神经元。本研究提供了基于质谱的人类诱导多能干细胞来源神经元在整个神经元分化周期中的定量蛋白质组特征谱，具体涵盖两类神经元：由诱导多能干细胞分化得到的诱导型谷氨酸能神经元与运动神经元。研究采用串联质谱标签10重标记技术，对不同时间点的细胞开展蛋白质组学表征分析。分析结果显示，在分化过程中多种关键蛋白的表达水平发生显著变化（错误发现率False Discovery Rate, FDR < 0.001），例如参与Wnt与Notch信号通路的蛋白。综上，本研究数据集为人类诱导多能干细胞源神经元分化过程中的蛋白质表达情况提供了丰富的信息资源。