Inhibition of Sirt1 promotes neural progenitors toward motoneuron differentiation from human embryonic stem cells.
Inhibition of Sirt1 promotes neural progenitors toward motoneuron differentiation from human embryonic stem cells.
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DOI：
10.1016/j.bbrc.2010.12.014
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发表时间：
2011-01
影响因子：
3.1
通讯作者：
Yun Zhang;Jing Wang;Gui-an Chen;Dongsheng Fan;M. Deng
中科院分区：
文献类型：
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作者：
Yun Zhang;Jing Wang;Gui-an Chen;Dongsheng Fan;M. Deng
有几种方案指导人类胚胎干细胞(HESCs)分化为功能性运动神经元，但运动神经元的生成效率因所用的人类ESC系而异。我们的目标是开发一种新的方案来增加人类ESCs运动神经元的形成。在这项研究中，我们测试了一种核组蛋白脱乙酰酶蛋白Sirt1，以促进人ESCs分化为运动神经元过程中神经前体细胞(NPC)的发育。Sirt1的特异性抑制剂烟酰胺显著增加运动神经元的形成。我们发现，大约60%的NPC细胞表达HB9和βⅢ-微管蛋白，这是在烟酰胺处理后的ESCs神经元中发现的常用的运动神经元标志物。ESC来源的运动神经元表达胆碱乙酰转移酶(ChAT)，这是成熟运动神经元的阳性标志。此外，我们还检测了Sirt1激活剂白藜芦醇(50μM)或抑制剂烟酰胺(100μM)对分化的NPC中Mash1、NGN2和HB9mRNA转录水平的影响。烟酰胺治疗后，Mash1、Ngn2和HB9mRNA的表达水平明显高于传统治疗方案的对照组。这些结果表明，通过抑制Sirt1来提高Mash1和Ngn2的水平可以提高HB9的表达，从而促进运动神经元的分化。这项研究为生产可移植的运动神经元提供了一种替代方法，这是发展基于hESC的运动神经元疾病细胞治疗的关键要求。
Several protocols direct human embryonic stem cells (hESCs) toward differentiation into functional motoneurons, but the efficiency of motoneuron generation varies based on the human ESC line used. We aimed to develop a novel protocol to increase the formation of motoneurons from human ESCs. In this study, we tested a nuclear histone deacetylase protein, Sirt1, to promote neural precursor cell (NPC) development during differentiation of human ESCs into motoneurons. A specific inhibitor of Sirt1, nicotinamide, dramatically increased motoneuron formation. We found that about 60% of the cells from the total NPCs expressed HB9 and βIII-tubulin, commonly used motoneuronal markers found in neurons derived from ESCs following nicotinamide treatment. Motoneurons derived from ESC expressed choline acetyltransferase (ChAT), a positive marker of mature motoneuron. Moreover, we also examined the transcript levels of Mash1, Ngn2, and HB9 mRNA in the differentiated NPCs treated with the Sirt1 activator resveratrol (50μM) or inhibitor nicotinamide (100μM). The levels of Mash1, Ngn2, and HB9 mRNA were significantly increased after nicotinamide treatment compared with control groups, which used the traditional protocol. These results suggested that increasing Mash1 and Ngn2 levels by inhibiting Sirt1 could elevate HB9 expression, which promotes motoneuron differentiation. This study provides an alternative method for the production of transplantable motoneurons, a key requirement in the development of hESC-based cell therapy in motoneuron disease.