ERK inhibition promotes neuroectodermal precursor commitment by blocking self-renewal and primitive streak formation of the epiblast.
ERK inhibition promotes neuroectodermal precursor commitment by blocking self-renewal and primitive streak formation of the epiblast.
复制标题
ERK 抑制通过阻止外胚层的自我更新和原条形成来促进神经外胚层前体定型
DOI：
10.1186/s13287-017-0750-8
复制
发表时间：
2018-01-05
影响因子：
7.5
通讯作者：
Li L
中科院分区：
文献类型：
--
作者：
Yu Y;Wang X;Zhang X;Zhai Y;Lu X;Ma H;Zhu K;Zhao T;Jiao J;Zhao ZA;Li L
背景多能干细胞在再生医学方面具有巨大的前景。然而，在临床应用之前，应建立可重复的多能干细胞分化方案。细胞外信号调节蛋白激酶（ERK）信号对于外胚层干细胞（EpiSC）的自我更新发挥着核心作用，但其在随后的胚层分化中的作用仍然不明确。我们提出ERK可以调节外胚层的分化。方法PD0325901用于抑制胚胎干细胞和EpiSC分化过程中的ERK激活。采用免疫荧光、蛋白质印迹分析、实时PCR和流式细胞术检测胚层标记和通路激活。结果我们证明小鼠E7.5胚胎神经外胚层中的ERK磷酸化水平低于原条中的ERK磷酸化水平。 ERK 抑制导致外胚层的神经谱系定型。从机制上讲，PD0325901 通过保留在细胞质中的 β-catenin 消除原条标记物的表达，并在 EpiSC 分化过程中抑制 OCT4 和 NANOG 的表达。因此，EpiSC 在 PD0325901 治疗下有效分化为神经外胚层谱系。这些结果表明，神经外胚层分化不需要外在信号，支持神经谱系的默认分化。结论我们报道，单一ERK抑制剂PD0325901可以将外胚层和EpiSC指定为神经样细胞，为神经分化提供了有效的策略。
BackgroundPluripotent stem cells hold great promise for regenerative medicine. However, before clinical application, reproducible protocols for pluripotent stem cell differentiation should be established. Extracellular signal-regulated protein kinase (ERK) signaling plays a central role for the self-renewal of epiblast stem cells (EpiSCs), but its role for subsequent germ layer differentiation is still ambiguous. We proposed that ERK could modulate differentiation of the epiblast.MethodsPD0325901 was used to inhibit ERK activation during the differentiation of embryonic stem cells and EpiSCs. Immunofluorescence, western blot analysis, real-time PCR and flow cytometry were used to detect germ layer markers and pathway activation.ResultsWe demonstrate that the ERK phosphorylation level is lower in neuroectoderm of mouse E7.5 embryos than that in the primitive streak. ERK inhibition results in neural lineage commitment of epiblast. Mechanistically, PD0325901 abrogates the expression of primitive streak markers by β-catenin retention in the cytoplasm, and inhibits the expression of OCT4 and NANOG during EpiSC differentiation. Thus, EpiSCs differentiate into neuroectodermal lineage efficiently under PD0325901 treatment. These results suggest that neuroectoderm differentiation does not require extrinsic signals, supporting the default differentiation of neural lineage.ConclusionsWe report that a single ERK inhibitor, PD0325901, can specify epiblasts and EpiSCs into neural-like cells, providing an efficient strategy for neural differentiation.
登录
查看更多内容
影响因子：
4.6
作者：
Dunn, NR;Vincent, SD;Bikoff, EK
通讯作者：
Bikoff, EK
影响因子：
64.8
作者：
Brons, I. Gabrielle M.;Smithers, Lucy E.;Vallier, Ludovic
通讯作者：
Vallier, Ludovic
影响因子：
4.8
作者：
Hayashida, T;deCaestecker, M;Schnaper, HW
通讯作者：
Schnaper, HW
影响因子：
4.6
作者：
Guo, Ge;Yang, Jian;Smith, Austin
通讯作者：
Smith, Austin
DOI：
10.1242/dev.156406
发表时间：
2017-10-15
期刊：
Development (Cambridge, England)
影响因子：
--
作者：
Piliszek A;Madeja ZE;Plusa B
通讯作者：
Plusa B