A phosphoproteomics study reveals a defined genetic program for neural lineage commitment of neural stem cells induced by olfactory ensheathing cell-conditioned medium
A phosphoproteomics study reveals a defined genetic program for neural lineage commitment of neural stem cells induced by olfactory ensheathing cell-conditioned medium
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DOI：
10.1016/j.phrs.2021.105797
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发表时间：
2021-08-11
影响因子：
9.3
通讯作者：
Lu, Ming
中科院分区：
文献类型：
--
作者：
Ge, Lite;Zhang, Cheng;Lu, Ming
由于嗅鞘细胞（OECs）和神经干细胞（NSC）在神经损伤和疾病的细胞治疗中均表现出一定的功效，近年来有一系列研究着眼于NSC和OECs的共培养。蛋白磷酸化形成了鉴定多种细胞信号传导途径的基础，这些细胞信号传导途径负责调节由OECs诱导的NSC的自我更新和分化。为了更好地理解在嗅鞘细胞诱导的神经干细胞分化的早期阶段的信号级联，在第一个24小时期间的神经干细胞蛋白质组和磷酸化蛋白质组的变化进行了测定，使用二甲基标记和二氧化钛磷酸化富集与液相色谱-串联质谱法（LC-MS/MS）。共鉴定出565个蛋白质和2511个磷酸化位点。根据OECs诱导NSC分化的前12和24 h的定量磷酸蛋白质组学分析，推测至少有两个不同的信号波：一个在刺激后12 h内达到峰值，第二个在24 h后达到峰值。除了了解蛋白质组和磷酸化蛋白质组在NSC分化早期阶段的动力学外，我们的分析还确定了OECs分泌的TGF-β 3蛋白的关键作用，其可能是促进OECs诱导的NSC分化为神经元的起始因子。这些发现不仅再次证明了在细胞治疗中基于OECs的治疗策略，而且还为OECs诱导的NSCs神经谱系定型的调节网络增加了一个节点。
Since both Olfactory ensheathing cells (OECs) and neural stem cells (NSCs) have shown certain efficacy in the cellular therapy of nerve injury and disease, there have been a series of investigations in recent years looking at the co-culture of NSCs and OECs. Protein phosphorylation forms the basis for identifying a variety of cellular signaling pathways responsible for regulating the self-renewal and differentiation of NSCs induced by OECs. To better understand the signaling cascades in the early phases of OEC-induced NSC differentiation, changes in the NSC proteome and phosphoproteome during the first 24 h were determined using dimethyl labeling and TiO2 phosphorylation enrichment coupled with Liquid chromatography-tandem mass spectrometry (LC-MS/MS). A total of 565 proteins and 2511 phosphorylation sites were identified. According to quantitative phosphoproteomics analyses of NSC differentiation induced by OECs during the first 12 and 24 h, it was speculated that there were at least two different signal waves: one peaking within 12 h after stimulation and the second upsurge after 24 h. In addition to understanding the dynamics of the proteome and phosphoproteome in the early stages of NSC differentiation, our analyses identified a key role of the TGF-beta 3 protein secreted by OECs, which may be an initiating factor that promotes differentiation of NSCs into neurons induced by OECs. These findings not only redemonstrated a OECs-based therapeutic strategy in cell therapy, but also added a node to the regulatory network for the neural lineage commitment of NSCs induced by OECs.