Functions of the ERK5 signalling pathway in Pluripotent Stem Cells

多能胚胎干细胞（ESC）具有自我更新和分化成成人体内所有特化细胞类型的能力。ESC的决策受到细胞信号网络的严格控制，我们的实验室试图解开并利用它来制定新的基于ESC的细胞替代策略（Fernando-Alonso et al，2017）。在最近的小分子激酶抑制剂筛选中，我们发现了ERK 5信号通路在控制ESC多能性中的关键功能。使用抑制剂工程和CRISPR/Cas9基因组编辑，我们发现，令人兴奋的是，ERK 5调节“幼稚”和“启动”多能ESC状态之间的转换（威廉姆斯等人，2016）。我们现在正在应用最先进的定量系统生物学技术来揭示ERK 5在ESCs中的分子功能。由于ERK 5独特地包含激酶结构域和推定的转录结构域，因此关键目标是其表达受ERK 5调控的基因。为此，我们将使用RNA-SEQ转录组学和总蛋白质组学分析质谱法来确定ERK 5在调节ESC基因表达和蛋白质组动力学中的作用。这揭示了ERK 5信号通路在调节早期胚胎基因表达中的新功能，这些基因与重新编程为幼稚多能性和获得全能性（一种发育不受限制的胚胎状态）相关。因此，我们要确定激活ESCs中ERK 5信号传导的分子机制，并利用这些信息将体细胞重编程为幼稚的多能性和潜在的全能性。最终，这方面的知识将提供新的途径，对细胞更有效地重新编程和totipotential. Keywords：胚胎干细胞，再生医学，信号转导，蛋白激酶定量技能：定量质谱，转录组学全生物体生理学：系统生物学，干细胞问题：1。解释跨学科接口：该项目通过采用定量质谱和转录组学分析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个生物体生理学相结合。项目是否需要大量的量化技能？3.项目是否需要大量的整体生物生理学技能？是的

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