Molecular feedback between cell division cycle and differentiation in pluripotent stem cells

多能干细胞细胞分裂周期和分化之间的分子反馈

• 批准号: 10502085
• 负责人: Ali Shariati
• 金额: $ 16.79万
• 依托单位: UNIVERSITY OF CALIFORNIA SANTA CRUZ
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10502085
• 关键词: Address; Automobile Driving; Biological; Biology; CRISPR screen; California; Cell Cycle; Cells; Collaborations; Complex; Engineering; Feedback; Genetic Engineering; Genetic Transcription; Goals; Label; Laboratories; Link; MEKs; Maintenance; Mitogen-Activated Protein Kinase Kinases; Mitogen-Activated Protein Kinases; Molecular; Outcome; Pathway interactions; Phosphotransferases; Pluripotent Stem Cells; Process; Public Health; Qi; Regenerative Medicine; Reproducibility; Testing; Therapeutic; Universities; cell type; chemical genetics; embryonic stem cell; experimental study; insight; novel; novel strategies; phosphoproteomics; pluripotency; professor; regeneration potential; response; self-renewal

Project Summary A major long-term goal of my laboratory is to determine molecular feedback mechanisms responsible for coordination between the cell division cycle and differentiation of embryonic stem cells (ESCs). ESCs have great therapeutic potential for regenerative medicine, because they can differentiate into any cell type and have unlimited self-renewal potential. This remarkable biological potential, known as pluripotency, is associated with a complex transcription network, an ultrafast cell division cycle that lacks typical checkpoints, and an atypical response to activation of the Mitogen-Activated Protein Kinase (MAPK) pathway. While a lot of progress has been made in characterizing the pluripotency transcription network, less is known about the cell division cycle of ESCs and its molecular links to the pluripotency transcription network. In addition, a critical yet poorly understood process is how ESCs use the MAPK pathway to control their self-renewal and differentiation. Our driving hypothesis is that the cell division cycle and pluripotency transcription network are linked through a bidirectional molecular feedback loop that is regulated by the MAPK activity. To test our hypothesis, we will: I. Characterize novel regulators of the MAPK pathway identified in our CRISPR screen in ESCs II. Determine functional substrate network of the MAPK kinases, Mek and Erk, in ESCs using chemical-genetic kinase engineering and quantitative phosphoproteomics. III. Determine mechanisms of pluripotency maintenance by the G1 cell cycle kinase (Cdk2) using direct labeling of the substrates by chemical-genetic engineering of Cdk2. To successfully complete proposed experiments, I have established collaborations with Professor Stanley Qi laboratory (Stanford University), Professor Alice Ting laboratory (Stanford University), Professor Seth Rubin laboratory (University of California, Santa Cruz) and Professor Boris Macek laboratory (University of Tubingen). By gaining detailed insight into the molecular mechanisms linking the cell division cycle and differentiation of ESCs, the outcome of this R35 proposal will provide novel strategies to address a key challenge in the field of regenerative medicine that is efficient and reproducible differentiation of ESCs for therapeutic purposes.

项目摘要 我的实验室的一个主要长期目标是确定分子反馈机制 负责协调胚胎干细胞的细胞分裂周期和分化 细胞(ESCs)。胚胎干细胞在再生医学方面具有巨大的治疗潜力，因为它们可以 分化为任何类型的细胞，并具有无限的自我更新潜力。这一非凡之处 生物潜能，即所谓的多能性，与复杂的转录网络有关， 缺乏典型检查点的超快细胞分裂周期，以及对激活的非典型反应 丝裂原活化蛋白激酶(MAPK)通路。虽然已经取得了很大的进展 在描述多能性转录网络时，人们对细胞分裂周期知之甚少 胚胎干细胞及其与多能性转录网络的分子链接。此外，一项关键的尚未 鲜为人知的过程是ESCs如何使用MAPK途径来控制其自我更新和 差异化。我们的驱动假设是细胞分裂周期和多能性 转录网络通过双向分子反馈环连接，即 受MAPK活性调控。为了检验我们的假设，我们将：I.描述新的监管者 我们CRISPR筛查发现的胚胎干细胞MAPK信号通路的研究II.功能底物的确定 利用化学遗传激酶工程研究胚胎干细胞中MAPK、MEK和ERK的网络 和定量磷酸蛋白质组学。三、通过以下方式确定多能性维持机制 利用化学遗传学直接标记底物的G1期细胞周期蛋白激酶(CDK2) CDK2的工程学。为了成功地完成拟议的实验，我已经建立了 与斯坦福大学实验室齐志鸿教授、丁雅玲教授合作 实验室(斯坦福大学)，Seth Rubin教授实验室(加州大学，圣塔市 和Boris Macek教授实验室(图宾根大学)。通过获得详细的洞察 关于连接胚胎干细胞分裂周期和分化的分子机制， R35提案的结果将提供新的战略，以应对这一领域的关键挑战 再生医学是胚胎干细胞有效和可重复分化的治疗方法 目的。