POST-TRANSCRIPTIONAL REGULATION OF CELL FATE IN EARLY MAMMALIAN DEVELOPMENT

早期哺乳动物发育中细胞命运的转录后调控

• 批准号: 10366001
• 负责人: CAROLYN O SANGOKOYA
• 金额: $ 14.24万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10366001
• 关键词: 3' Untranslated Regions; Address; Anatomy; Binding Proteins; Biosensor; CRISPR/Cas technology; Cell Differentiation process; Cell Fate Control; Cell model; Cells; Critical Pathways; Data; Development; Development Plans; Developmental Biology; Embryo; Embryonic Development; Endocytosis; Ensure; Environment; Erythropoiesis; Family; Fibroblast Growth Factor; Five-Year Plans; Future; Goals; Grant; Hepatobiliary; High-Throughput Nucleotide Sequencing; Human Development; Immunoprecipitation; In Vitro; Iron; Iron Regulatory Protein 1; Iron-Regulatory Proteins; Knowledge; Laboratories; Leadership; Learning; Maps; Mediating; Mentored Clinical Scientist Development Award (K08); Mentors; Messenger RNA; Methods; MicroRNAs; Modeling; Molecular; Nucleotides; Operative Surgical Procedures; Organogenesis; Pathologist; Pathology; Pathway interactions; Pattern; Physicians; Play; Post-Transcriptional Regulation; Process; RNA-Binding Proteins; Regulation; Reporter; Research; Resolution; Role; Scientist; Signal Transduction; Site; Statistical Methods; Stem cell pluripotency; Technical Expertise; Testing; Training; Transcript; base; career development; crosslink; embryonic stem cell; functional genomics; gastrointestinal; gastrulation; in vivo; mouse model; multidisciplinary; mutant; neurogenesis; pluripotency; profilin; regenerative therapy; single-cell RNA sequencing; skills; stem; stem cell biology; stem cell differentiation; stem cell fate; stem cell self renewal; stem cells; tool; transcriptome

Post-transcriptional regulation by RNA-binding proteins (RBPs) and microRNAs (miRNAs) orchestrate diverse molecular and cellular mechanisms that pattern early mammalian development from embryonic stem cells (ESCs) through gastrulation and lineage commitment. The RBPs Ago2 (Argonaute-2) and IRP (iron regulatory proteins) respectively coordinate miRNA-mediated regulation and cellular iron regulation, mechanisms essential for the proper execution of early embryonic development. In human development, cellular iron regulation is important for non-hematopoietic tissue development, including neurogenesis and gut development, in addition to erythropoiesis. However, the identities and functional roles of miRNA- and IRP-bound targets in cell fate decisions during early embryonic development are largely unknown. A comprehensive understanding of the dynamic relationships of IRPs, miRNAs, and their functional targets during this critical developmental window is needed and can provide a roadmap for functional rewiring in stem/progenitor cell-based regenerative therapies. The central hypothesis of this proposal is that IRP and miRNAs function cooperatively and dynamically on targets that are important regulators of cell fate transitions during mammalian development. As a molecular biologist and pathologist, my long-term goal is to understand and use these integrated pathways of post-transcriptional control to devise new tools and approaches for functional rewiring in stem/progenitor cell-based regenerative therapies. The objective of this project is to dissect the specific roles of post-transcriptional regulation by miRNAs and IRPs on cell fate decisions in early mammalian development and to build platforms to model cellular iron throughout early mammalian development. This project objective will be achieved by 1) determining the impact of IRP and miR-290-mediated regulation on Profilin-2, a known regulator of ESC differentiation, 2) identifying and functionally dissecting the global network of bound IRP and miRNA targets, and 3) developing biosensor platforms that model cellular iron utilization in vitro and in vivo during early embryonic development. The proposed studies are the core components of the Mentored Clinical Scientist Development Award (K08) for Dr. Carolyn Sangokoya. Dr. Sangokoya is a board-certified Anatomic Pathologist with subspeciality expertise in surgical and gastrointestinal/hepatobiliary pathology. This proposal encompasses a five-year plan to address gaps in specific research and professional skills as she transitions to independence as a physician-scientist. This grant is a training vehicle for Dr. Sangokoya to 1) build knowledge in statistical methods for functional genomics, 2) learn and expand technical skills in generating mouse models, 3) perform single-cell RNA-sequencing studies, and 4) develop professional scientific leadership and lab management skills in transition to leading a successful laboratory. To achieve these goals, Dr. Sangokoya and her multidisciplinary scientific advisory and mentoring team have devised a 5-year career development plan. The proposed training, didactics, and research in the rich research environment at UCSF will ensure a successful and productive transition to independence.

RNA 结合蛋白 (RBP) 和 microRNA (miRNA) 的转录后调控协调多种 胚胎干细胞早期哺乳动物发育模式的分子和细胞机制 （ESC）通过原肠胚形成和谱系定型。 RBP Ago2 (Argonaute-2) 和 IRP (铁监管 蛋白质）分别协调 miRNA 介导的调节和细胞铁调节，机制至关重要 为了正确执行早期胚胎发育。在人类发育中，细胞铁调节是 此外，对非造血组织发育很重要，包括神经发生和肠道发育 到红细胞生成。然而，miRNA 和 IRP 结合靶标在细胞命运中的身份和功能作用 早期胚胎发育过程中的决定很大程度上是未知的。全面了解 在这个关键的发育窗口期间，IRP、miRNA 及其功能靶标的动态关系是 需要并可以为基于干细胞/祖细胞的再生疗法中的功能重新布线提供路线图。 该提案的中心假设是 IRP 和 miRNA 在目标上协同动态地发挥作用 它们是哺乳动物发育过程中细胞命运转变的重要调节因子。作为一名分子生物学家 和病理学家，我的长期目标是理解和使用转录后的这些整合途径 控制设计新的工具和方法，用于基于干细胞/祖细胞的再生功能重新布线 疗法。该项目的目的是剖析 miRNA 转录后调控的具体作用 和 IRP 对早期哺乳动物发育中细胞命运决定的影响，并构建细胞铁模型平台 整个早期哺乳动物的发育过程。该项目目标将通过以下方式实现：1) 确定影响 IRP 和 miR-290 介导的对 Profilin-2 的调节，Profilin-2 是 ESC 分化的已知调节因子，2) 识别 并在功能上剖析绑定 IRP 和 miRNA 靶标的全局网络，以及 3) 开发生物传感器 模拟早期胚胎发育过程中体外和体内细胞铁利用的平台。这 拟议的研究是 Dr. 指导临床科学家发展奖 (K08) 的核心组成部分。 卡罗琳·桑戈科亚。 Sangokoya 博士是一位经过委员会认证的解剖病理学家，拥有以下领域的专科专业知识： 外科和胃肠道/肝胆病理学。该提案包含一个五年计划，旨在解决 当她过渡为独立的医师科学家时，她在具体研究和专业技能方面存在差距。这 赠款是 Sangokoya 博士的培训工具，旨在 1）建立功能基因组学统计方法方面的知识， 2) 学习并扩展生成小鼠模型的技术技能，3) 进行单细胞 RNA 测序研究， 4) 培养专业的科学领导力和实验室管理技能，以引领成功的转型 实验室。为了实现这些目标，Sangokoya 博士和她的多学科科学咨询和指导 团队制定了5年职业发展规划。拟议的针对富人的培训、教学和研究 加州大学旧金山分校的研究环境将确保向独立的成功和富有成效的过渡。