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Determining feedback mechanisms between cell cycle and cell fate in pluripotent cells

确定多能细胞中细胞周期和细胞命运之间的反馈机制

基本信息

批准号：
10437621
负责人：
Ali Shariati
金额：
$ 24.9万
依托单位：
UNIVERSITY OF CALIFORNIA SANTA CRUZ
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：
2018-07-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10437621
关键词：
Attention Automobile Driving Award Binding Binding Sites Biochemical Biological Biological Sciences Biology CRISPR/Cas technology Cell Cycle Cell Cycle Progression Cell Cycle Regulation Cell Lineage Cell division Cells ChIP-seq Chemicals Collaborations Complement Factor B Complex Conflict (Psychology)Cyclins Disease ES Cell Line Engineering Ensure Feedback Future Teacher G1 Phase G1/S Checkpoint Pathway Genetic Genetic Transcription Germany Goals Growth Factor In Vitro Interdisciplinary Study Link MEKKs MEKs Maintenance Measures Medicine Methods Mitogen-Activated Protein Kinases Mitogens Modeling Molecular Pathway interactions Phase Phosphorylation Site Phosphotransferases Pluripotent Stem Cells Process Protein Engineering Public Health Qi Regenerative Medicine Regulation Reporter Research Research Personnel Research Project Grants Role Series Signal Transduction Somatic Cell Stem cell pluripotency Testing Training Universities analog career cell type embryonic stem cell experimental study factor C genome-wide in vivo insight novel novel therapeutic intervention phosphoproteomics pluripotency pluripotency factor promoter quantitative imaging response self-renewal transcription factor upstream kinase virtual

项目摘要

Project Summary/Abstract Embryonic stem cells (ESCs) hold great promise for medicine because they can be propagated to virtually unlimited numbers and can generate any disease relevant cell type. ESCs have three unique cell biological features that make them distinct from somatic cell lineages: (i) A pluripotency transcriptional network that promotes its own activity; (ii) An atypically rapid cell cycle with short G1 phase that lacks a typical restriction point; (iii) An atypical response to Mitogen Activated Protein Kinase (MAPK) activity1–3. While a lot of attention has focused on the maintenance of ESC pluripotency by a transcriptional network, research on the ESC cell cycle network has been largely descriptive, and the mechanistic links between the two networks have yet to be explored. In addition, a critical but poorly understood process is how ESCs use MAPK pathway to control the exit from pluripotency. My long-term career goal is to discover the molecular mechanisms that allows ESCs to choose between two conflicting fates, i.e, self-renewal vs. fate commitment. The driving hypothesis of this proposal is that the ESC-specific cell cycle is functionally linked with the transcriptional pluripotency network by mutual, positive feedback that is regulated by upstream activity of MAPK signaling. To test this hypothesis, we propose to investigate the function of phosphorylation sites on the pluripotency factors using both genetic and biochemical methods (Aim1). We will employ single cell quantitative imaging to measure dynamics of cell cycle in ESCs expressing reporters of pluripotency and cell cycle. To test if pluripotency factors directly promote cell cycle progression, we are proposing to develop a novel method by repurposing the CRISPR/Cas9 technology to examine the function of specific transcription factor binding sites in vivo (Aim2). To determine how MAPK signaling regulates exit from pluripotency, we will combine protein engineering and quantitative phosphoproteomics to uncover novel targets of this pathway (Aim3). Completion of these aims will reveal the mechanisms by which ESCs choose between opposing fates, i.e. self-renewal vs. fate commitment. During the training phase of this award (K99), I plan to leverage quantitative insight of Skotheim’s lab to advance an interdisciplinary research plan to study cell fate in pluripotent cells in my lab. To this end, I have established collaboration with Wernig’s lab (Stanford), Qi’s lab(Stanford) and Macek’s lab(Tubingen University, Germany) that will greatly facilitate the progress of my research project. To prepare for transition to an independent investigator, I will take part in career training courses such as The Future Faculty Series that are offered by Stanford University. In addition to representing an important advance in basic biological sciences, our mechanistic insight may facilitate propagation and lineage differentiation of ESCs for regenerative medicine.

项目总结/文摘