The National & Local Joint Engineering Laboratory of Animal Peptide Drug Development, College of Life Sciences, Hunan Normal University, Changsha 410006, PR China; Department of Neurology, Second Xiangya Hospital, Central South University, Changsha 410011, PR China; Hunan Provincical Key Laboratory of Neurorestoratology, The Second Affiliated Hospital, Hunan Normal University, Changsha 410003, PR China.
The National & Local Joint Engineering Laboratory of Animal Peptide Drug Development, College of Life Sciences, Hunan Normal University, Changsha 410006, PR China; Guangdong Provincial Key Laboratory of Biotechnology for Plant Development, School of Life Science, South China Normal University, Guangzhou 510631, PR China.
Pharmacol Res. 2021 Oct;172:105797. doi: 10.1016/j.phrs.2021.105797. Epub 2021 Aug 2.
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 TiO 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-β3 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.
近年来，由于嗅鞘细胞（OEC）和神经干细胞（NSC）在神经损伤和疾病的细胞治疗中都显示出一定的疗效，因此进行了一系列将 NSC 和 OEC 共培养的研究。蛋白质磷酸化构成了鉴定各种细胞信号通路的基础，这些信号通路负责调节 OEC 诱导的 NSC 的自我更新和分化。为了更好地了解 OEC 诱导 NSC 分化的早期阶段的信号级联反应，使用二甲标记和 TiO2 磷酸化富集与液相色谱-串联质谱（LC-MS/MS）联用，在最初的 24 小时内确定了 NSC 蛋白质组和磷酸蛋白质组的变化。共鉴定到 565 种蛋白质和 2511 个磷酸化位点。根据 OEC 诱导 NSC 在最初 12 小时和 24 小时分化过程中 12 小时和 24 小时的定量磷酸化蛋白质组学分析，推测至少存在两个不同的信号波：一个在刺激后 12 小时内达到峰值，另一个在 24 小时后再次增加。除了了解 NSC 分化早期蛋白质组和磷酸蛋白质组的动态变化外，我们的分析还确定了 OEC 分泌的 TGF-β3 蛋白的关键作用，它可能是促进 OEC 诱导的 NSCs 向神经元分化的起始因子。这些发现不仅在细胞治疗中再次证明了基于 OEC 的治疗策略，而且为 OEC 诱导的 NSCs 向神经谱系分化的调控网络增加了一个节点。
