神经干细胞(NSCs)增殖、分化命运转变与决定机制是NSCs基础研究中最重要的科学问题。本课题在前期研究中发现一种小分子化合物在NSCs分化命运转换过程中发挥重要作用，明显抑制NSCs分化，并首次发现其调控重要表观遗传因子CBP/P300乙酰化转移酶活性这一具有重要研究价值的现象。本研究拟在前期研究基础上，首先利用分子生物学和细胞组织学等技术检测NSCs的分化命运，及多潜能因子、命运决定因子和细胞周期调节因子的变化；再利用透射电镜和染色质免疫共沉淀等技术研究染色质重塑、组蛋白修饰变化及其相关转录因子的表达，并寻找调控CBP/P300乙酰化转移酶活性的具体分子机制。本研究建立了多潜能因子、命运决定因子、表观遗传因子及蛋白修饰等在胚胎NSCs分化命运转换阶段的相互作用网络，加深了对胚胎NSCs发育信号分子调控网络的理解，并使得未来通过使用小分子化合物改变细胞命运并且重塑所需细胞的功能成为可能。

The mechanism of proliferation, differentiation as well as the fate switch process between proliferation and differentiation is of great importance in the study of neural stem cells (NSCs). We recently found the role of a small-molecule compound in the process of fate switch between self-renewal and differentiation in NSCs. The small-molecule compound significantly inhibited the differentiation of NSCs and more interestingly，regulated the histone acetyltransferase activity of CBP/P300. In this connection， further studies are designed to explore the function and regulatory mechanism of this small-molecule compound in NSCs. First, the effects of this small-molecule compound treatment on the fate determination, expression of stemness-related gene and cell cycle regulators in NSCs will be determined using the molecular biological and cytohistological techniques. Secondly, TEM and ChIP will be used to study the changes in chromatin remodeling, histone modification and expression of related transcriptional factors. Finally, the exact molecular target in regulating CBP/P300 will be investigated. Our study will establish the net by elucidating the relationship among multipotency factors, fate determination factors, epigenetic regulators and histone modification in the fate switch in NSCs. This study will contribute to understand the mechanism of NSCs differentiation and provide a new insight into the regulation of NSCs fate, which may benefit the strategy of controlling cell fate switch and lineage reprogramming by small-molecule compound.

神经干细胞（NSCs）的增殖、分化是神经系统正常发育的细胞基础，如何准确的调控NSCs增殖、分化命运的转换是NSCs基础研究中最重要的科学问题。本项目在成功建立体外分离培养NSCs的细胞模型的基础上，利用Western blot、免疫荧光染色、FACS、透射电镜、染色质免疫共沉淀等多种形态学和分子生物学方法，对棕榈酰转移酶抑制剂——小分子化合物2BRO，在神经发育中的作用及分子机制进行了研究。我们研究发现2BRO通过抑制重要表观遗传学因子CBP/p300的组蛋白乙酰转移酶活性，降低转录因子启动子区域组蛋白H3的乙酰化水平，从而阻碍了NSCs的分化。本研究建立了分化命运决定因子、表观遗传因子及蛋白修饰等在胚胎NSCs分化命运转换阶段的相互作用网络，加深了对NSCs发育信号分子调控网络的理解，并使得未来通过使用小分子化合物改变细胞命运并且重塑所需细胞的功能成为可能。