Functions of the ERK5 signalling pathway in Pluripotent Stem Cells

多能胚胎干细胞(ESCs)具有自我更新和分化为成人体内所有特化细胞类型的能力。ESC的决策受到蜂窝信号网络的严格控制，我们的实验室试图解开并利用这些网络来制定基于ESC的新型细胞替换策略(Fernandez-Alonso等人，2017)。在最近的一次小分子激酶抑制剂筛查中，我们发现了ERK5信号通路在控制ESC多能性方面的关键功能。利用抑制剂工程和CRISPR/Cas9基因组编辑，我们令人兴奋地发现，ERK5调节“幼稚”和“启动”多能ESC状态之间的转换(Williams等人，2016)。我们现在正在应用尖端的定量系统生物学技术来解开ERK5在ESCs中的分子功能。由于ERK5独特地含有一个激酶域和一个假定的转录结构域，因此一个关键的目的是寻找其表达受ERK5调控的基因。为此，我们将利用RNA-SEQ转录组学和总蛋白质组谱质谱学来确定ERK5在调节ESC基因表达和蛋白质组动力学中的作用。这揭示了ERK5信号通路在调节早期胚胎基因表达方面的新功能，这些基因与重新编程为天真的多能性和达到全能性相关，全能性是一种发育不受限制的胚胎状态。因此，我们需要确定在ESCs中激活ERK5信号的分子机制，并利用这一信息重新编程体细胞，使其具有天真的多能性和潜在的全能性。关键词：胚胎干细胞，再生医学，信号转导，蛋白质运动定量技能：定量质谱学，转录学全球生物生理学：系统生物学，干细胞问题：1.解释跨学科接口：这个项目通过使用定量质谱学和转录图谱来解决干细胞中的系统生物学问题，将定量分析与系统生物学和整体生物体生理学结合起来。项目是否需要大量的量化技能？是的3.项目需要大量的整体生理技能吗？是

Pluripotent Embryonic Stem Cells (ESCs) have the capacity to self-renew and differentiate into all specialized cell types in the adult body. ESC decision-making is tightly controlled by cellular signalling networks, which our lab seeks to unravel and exploit to elaborate novel ESC-based cell replacement strategies (Fernandez-Alonso et al, 2017). In a recent small molecule kinase inhibitor screen, we uncovered a critical function for the ERK5 signalling pathway in controlling ESC pluripotency. Using inhibitor engineering and CRISPR/Cas9 genome editing, we find, excitingly, that ERK5 modulates transition between "naïve" and "primed" pluripotent ESC states (Williams et al, 2016). We are now applying cutting-edge quantitative systems biology technologies to unravel molecular functions of ERK5 in ESCs. As ERK5 uniquely contains a kinase domain and a putative transcriptional domain, a key aim is to gene whose expression is regulated by ERK5. To this end, we will define the role of ERK5 in regulating ESC gene expression and proteome dynamics using RNA-SEQ transcriptomics and total proteomic profiling mass-spectrometry. This uncovers a new function for the ERK5 signalling pathway in regulating expression of early embryonic genes associated with reprogramming to naïve pluripotency and attaining totipotency, an embryonic state that is developmentally unrestricted. Therefore, we to identify molecular mechanisms that activate ERK5 signalling in ESCs, and exploit this information to reprogram somatic cells to naïve pluripotency and potentially totipotency. Ultimately, this knowledge will provide new avenues towards reprogramming cells more efficiently and towards totipotency.Key Words: Embryonic Stem Cell, Regenerative Medicine, Signal Transduction, Protein KinaseQuantitative Skills: Quantitative Mass-Spectrometry, TranscriptomicsWhole Organism Physiology: Systems Biology, Stem CellsQuestions:1. Explain interdisciplinary interface: This project interfaces quantitative analysis with systems biology and whole organism physiology by employing quantitative mass-spectrometry and transcriptomic profiling to address systems biology question in stem cells2. Does project require significant amount of quantitative skills? YES 3. Does project require significant amount of whole organism physiology skills? YES