Quantitative proteomics analysis highlights the role of redox hemostasis and energy metabolism in human embryonic stem cell differentiation to neural cells
Quantitative proteomics analysis highlights the role of redox hemostasis and energy metabolism in human embryonic stem cell differentiation to neural cells
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DOI：
10.1016/j.jprot.2014.02.002
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发表时间：
2014-04-14
影响因子：
3.3
通讯作者：
Salekdeh, Ghasem Hosseini
中科院分区：
文献类型：
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作者：
Fathi, Ali;Hatami, Maryam;Salekdeh, Ghasem Hosseini
人类胚胎干细胞（hESCs）的神经分化是体外分析人类神经发生的独特机会。通过神经板形成的外部线索在hESCs中得到了很好的描述，尽管神经发育的细胞内机制在很大程度上是未知的。hESC向神经细胞分化的蛋白质组学分析将有助于进一步确定人类神经发生的分子机制。利用二维凝胶电泳（2D-DIGE）系统分析了hESC向神经元分化的三个阶段，即早期神经分化、神经外胚层和成熟神经元的蛋白质组。利用MALDI-TOF/TOF和LC - MS/MS鉴定了137个差异积累蛋白点中的118个。我们观察到，参与氧化还原止血、维生素和能量代谢以及泛素依赖性蛋白质水解的蛋白质在分化细胞中更丰富，而与RNA加工和蛋白质折叠相关的蛋白质在hESCs中含量更高。参与维持细胞氧化还原状态的蛋白质丰度较高，表明氧化还原止血在神经分化中的重要性。此外，我们的研究结果支持神经元活动和葡萄糖利用之间耦合机制的概念。蛋白质网络分析表明，大多数相互作用蛋白与细胞周期和细胞增殖有关。这些结果增强了我们对神经承诺和分化的分子动力学的理解。通过强调神经细胞氧化还原的作用和独特的代谢特性，本研究结果为我们对hESC向神经元分化的理解提供了新的见解。14种参与维持细胞氧化还原状态的蛋白，包括10个过氧化物还氧蛋白（Prclx）家族成员，其丰度主要在分化过程中增加，从而突出了神经分化与氧化还原的联系。我们的研究结果显示，在分化过程中，编码糖酵解和氨基酸合成酶的基因的表达明显增加。蛋白质网络分析预测了一些hESC分化的关键介质。这些蛋白包括TP53， CTNNB1, SMARCA4， TNF， TERT, E2F1， MYC， RB1和AR。(C) 2014 Elsevier B.V.版权所有。
Neural differentiation of human embryonic stem cells (hESCs) is a unique opportunity for in vitro analyses of neurogenesis in humans. Extrinsic cues through neural plate formation are well described in the hESCs although intracellular mechanisms underlying neural development are largely unknown. Proteome analysis of hESC differentiation to neural cells will help to further define molecular mechanisms involved in neurogenesis in humans. Using a two-dimensional differential gel electrophoresis (2D-DIGE) system, we analyzed the proteome of hESC differentiation to neurons at three stages, early neural differentiation, neural ectoderm and mature neurons. Out of 137 differentially accumulated protein spots, 118 spots were identified using MALDI-TOF/TOF and LC MS/MS. We observed that proteins involved in redox hemostasis, vitamin and energy metabolism and ubiquitin dependent proteolysis were more abundant in differentiated cells, whereas the abundance of proteins associated with RNA processing and protein folding was higher in hESCs. Higher abundance of proteins involved in maintaining cellular redox state suggests the importance of redox hemostasis in neural differentiation. Furthermore, our results support the concept of a coupling mechanism between neuronal activity and glucose utilization. The protein network analysis showed that the majority of the interacting proteins were associated with the cell cycle and cellular proliferation. These results enhanced our understanding of the molecular dynamics that underlie neural commitment and differentiation.Biological significance In highlighting the role of redox and unique metabolic properties of neuronal cells, the present findings add insight to our understanding of hESC differentiation to neurons. The abundance of fourteen proteins involved in maintaining cellular redox state, including 10 members of peroxiredoxin (Prclx) family, mainly increased during differentiation, thus highlighting a link of neural differentiation to redox. our results revealed markedly higher expression of genes encoding enzymes involved in the glycolysis and amino acid synthesis during differentiation. Protein network analysis predicted a number of critical mediators in hESC differentiation. These proteins included TP53, CTNNB1, SMARCA4, TNF, TERT, E2F1, MYC, RB1, and AR. (C) 2014 Elsevier B.V. All rights reserved.