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（miRNAs）的转录后调控协调了多种 从胚胎干细胞到哺乳动物早期发育的分子和细胞机制 （ESC）通过原肠胚形成和谱系定型。RBP Ago 2（Argonaute-2）和IRP（铁调节蛋白） 蛋白质）分别协调miRNA介导的调节和细胞铁调节， 以确保早期胚胎发育的正确执行。在人类发育中，细胞铁调节是 对非造血组织发育，包括神经发生和肠道发育很重要，此外 到红细胞生成然而，miRNA和IRP结合靶点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在很大程度上是未知的。全面了解 在这个关键的发育窗口期，IRP、miRNA及其功能靶点之间的动态关系， 需要，并可以提供一个路线图功能重新布线在干/祖细胞为基础的再生疗法。 该建议的中心假设是IRP和miRNAs在靶点上协同动态地发挥作用 是哺乳动物发育过程中细胞命运转变的重要调节因子。作为一个分子生物学家 作为一名病理学家，我的长期目标是了解和利用这些转录后整合途径， 控制设计新的工具和方法，用于干/祖细胞为基础的再生 治疗本项目的目的是研究miRNAs在转录后调控中的具体作用 和IRP在早期哺乳动物发育中的细胞命运决定，并建立平台来模拟细胞铁 在哺乳动物的早期发育中。本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（一种已知的ESC分化调节因子）的调节，2）鉴定 并从功能上剖析结合的IRP和miRNA靶标的全球网络，以及3）开发生物传感器 在早期胚胎发育过程中体外和体内模拟细胞铁利用的平台。的 拟议的研究是指导临床科学家发展奖（K 08）的核心组成部分，博士。 卡罗琳·桑戈科娅Sangokoya博士是一名经过委员会认证的解剖病理学家，拥有以下专业知识： 外科和胃肠道/肝胆病理学。该提案包括一个五年计划，以解决 在具体的研究和专业技能的差距，因为她过渡到独立作为一个医生，科学家。这 赠款是Sangokoya博士的培训工具，用于1）建立功能基因组学统计方法的知识， 2)学习和扩展建立小鼠模型技术技能，3）进行单细胞RNA测序研究， 和4）发展专业的科学领导和实验室管理技能， 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 团队制定了五年职业发展计划。建议的培训，教学法，并在丰富的研究 在UCSF的研究环境将确保成功和富有成效的过渡到独立。