Embryonic stem cell self-renewal

胚胎干细胞自我更新

基本信息

批准号：
9912157
负责人：
Qilong Ying
金额：
$ 33万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-01 至 2022-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9912157
关键词：
Affect Amino Acids Biologic Development Cell Culture Techniques Cell Nucleus Cells Chemicals Cytoplasm Data Development Disease Disease model Extracellular Signal Regulated Kinases Functional disorder Gene Deletion Glycogen Synthase Kinase 3 Goals Gold Human Individual Inner Cell Mass MAPK1 gene MAPK3 gene Mammals Mediating Mitogen-Activated Protein Kinase Kinases Modeling Molecular Mus Pathologic Phenotype Phosphotransferases Physiological Play Pluripotent Stem Cells Process Protein Kinase Proteins Rattus Regenerative Medicine Research Role Signal Pathway Signal Transduction Technology Testing Time Work base beta catenin cell type chemical genetics embryonic stem cell gene function genetic approach human disease human model inhibitor/antagonist innovation insight kinase inhibitor member natural Blastocyst Implantation novel paralogous gene preimplantation scaffold self-renewal small molecule stem cell fate success therapy development tool transmission process

项目摘要

Embryonic stem cells (ESCs) are pluripotent stem cells derived from the inner cell mass of the pre-implantation blastocyst. ESCs provide a powerful platform for elucidating gene function and creating disease models. So far, the application of ESC-based technologies has been limited to mice and rats, because germline competent ESCs have not yet been established from non-rodent species. Our previous work established that efficient self- renewal of mouse and rat ESCs could be achieved by dual inhibition of glycogen synthase kinase 3 (GSK3) and mitogen-activated protein kinase kinase (MEK). GSK3 has two paralogous members, GSK3α and GSKβ, which share nearly identical kinase domains. Similarly, extracellular signal-regulated kinase (ERK), the only known physiological substrates of MEK, also has two highly homologous paralogs, ERK1 and ERK2. Despite their high levels of homology, GSK3α and GSK3β possess distinct functions in regulating ESC self-renewal; the same is the case for ERK1 and ERK2. Therefore, fine-tuning of GSK3 and ERK signaling becomes critical in achieving optimal ESC self-renewal. By taking advantage of the chemical-genetic approach and the ESC platform, we propose to investigate the molecular mechanisms underlying the distinct functions of individual GSK3 and ERK paralogs in ESC self-renewal. In Aim 1, we will identify and characterize GSK3 paralog- specific substrates and downstream targets. We will also determine the amino acid variances responsible for the distinct functions of GSK3α and GSK3β. In Aim 2, we will identify and characterize ERK paralog-specific substrates in the cytoplasm and nucleus and investigate how subcellular localization of ERK1 and ERK2 affects their functions in ESCs. In Aim 3, we will fine-tune GSK3 and ERK signaling in mouse and human naïve ESCs to achieve optimal self-renewal effect. Success with these three aims will not only provide insights into the molecular basis of ESC self-renewal, but will also have far-reaching implications for our deep understanding of pathological conditions caused by dysfunction of GSK3 and ERK.

胚胎干细胞（ESC）是源自植入前内细胞团的多能干细胞迄今为止，胚胎干细胞为阐明基因功能和创建疾病模型提供了强大的平台。基于 ESC 的技术的应用仅限于小鼠和大鼠，因为种系具有能力我们之前的工作尚未从非啮齿动物物种中建立起高效的自体胚胎干细胞。小鼠和大鼠 ESC 的更新可以通过糖原合成酶激酶 3 (GSK3) 的双重抑制来实现丝裂原激活蛋白激酶激酶 (MEK) 有两个旁系同源成员：GSK3α 和 GSKβ，类似地，细胞外信号调节激酶（ERK）具有几乎相同的激酶结构域。尽管已知 MEK 的生理底物，但也有两个高度同源的旁系同源物：ERK1 和 ERK2。 GSK3α和GSK3β具有高度同源性，在调节ESC自我更新方面具有不同的功能； ERK1和ERK2的情况也是如此，因此，GSK3和ERK信号的微调变得至关重要。通过利用化学遗传方法和 ESC 来实现最佳的 ESC 自我更新。平台，我们建议研究个体不同功能背后的分子机制 ESC 自我更新中的 GSK3 和 ERK 旁系同源物在目标 1 中，我们将识别并表征 GSK3 旁系同源物。我们还将确定造成的氨基酸差异。 GSK3α 和 GSK3β 的不同功能在目标 2 中，我们将识别并表征 ERK 旁系同源物特异性。细胞质和细胞核中的底物并研究 ERK1 和 ERK2 的亚细胞定位影响它们在 ESC 中的功能在目标 3 中，我们将在小鼠和人类实验中微调 GSK3 和 ERK 信号传导。 ESC 要想成功实现这三个目标，不仅可以提供深入的见解。 ESC自我更新的分子基础，但也将对我们的深层产生深远的影响了解 GSK3 和 ERK 功能障碍引起的病理状况。