The Role of NMD in Cortical Neural Progenitor Cells

NMD 在皮质神经祖细胞中的作用

• 批准号: 10572698
• 负责人: Sika Zheng
• 金额: $ 3.13万
• 依托单位: UNIVERSITY OF CALIFORNIA RIVERSIDE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10572698
• 关键词: Ablation; Affect; Award; Behavior; Brain; California; Cell Cycle; Cell Cycle Regulation; Cell Lineage; Cells; Cellular biology; Clustered Regularly Interspaced Short Palindromic Repeats; Cortical Malformation; Data; Defect; Development; Disease; Doctor of Philosophy; Ensure; Etiology; Funding; GADD45B/GADD45G; Genetic; Goals; Knockout Mice; Knowledge; Laboratories; Lead; Light; Macrocephaly; Mediating; Mentorship; Messenger RNA; Microcephaly; Molecular; Molecular Biology; Mutation; Neocortex; Neurodevelopmental Disorder; Neuroglia; Neurons; Neurosciences; Nonsense Codon; Output; Parents; Pathogenicity; Phenotype; Process; RNA Decay; RNA Stability; Radial; Regulation; Research; Research Training; Role; Science; Structure; Testing; Time; Training; Transcript; Universities; career; cell behavior; confocal imaging; developmental neurobiology; doctoral student; in vivo; insight; mRNA Decay; mRNA Stability; mouse genetics; nerve stem cell; neurodevelopment; neurogenesis; neuroregulation; novel; progenitor; relating to nervous system; stem; stem cells; transcriptomics

PROJECT SUMMARY Mutations in key factors of nonsense-mediated mRNA decay (NMD), including Upf2, Upf3a, Upf3b, and Smg6, are enriched in various neurodevelopmental diseases. In additional to ensuring transcript quality by degrading aberrant transcripts with a premature stop codon, NMD modulates stability of selective mRNAs to fine-tune transcript abundance. Whether and how NMD influences brain development remains elusive. Our long-term objective is to understand the functional role of NMD regulation for the complicated and dynamic process of neurogenesis and how its mis-regulation leads to neurodevelopmental disorders. We determine the requirement of NMD for neural development through selective genetic ablation of Upf2 and in vivo manipulation of other NMD factors. Our preliminary data show that deletion of UPF2 in neural stem and progenitors results in microcephaly. UPF2 loss specifically affects the cell cycle and lineage progression of radial glia cells (RGCs), the major neural progenitor cells in the developing neocortex. We will combine cutting edge molecular cellular ribogenomics approaches, mouse genetics, and developmental neurobiology to dissect the mechanisms of NMD regulating neurogenesis. We propose three independent and interrelated aims to investigate possible variables underlying the microcephaly phenotype. In Aim 1, we will determine the cell cycle behaviors of RGCs in NMD knockout mice qualitatively and quantitatively and unveil the underlying regulatory mechanisms. In Aim 2, we will determine the lineage progression of RGCs and the resulting neuronal outputs per time unit in NMD knockout mice. By characterizing these molecular cellular defects, we also aim to provide mechanistic insights to transcriptomic regulation of RGC’s lineage transitions. NMD may regulate cell fates either independent of cell cycle controls or as the consequence of affecting the cell cycle. In Aim 3, we will test these two hypotheses and leverage our results to reexamine the relationship between cell cycle and cell fate. Successful completion of these studies will provide fundamental insights into how selective mRNA stability underlies the highly regulated cortical neurogenesis process in the mammalian brain. The proposed studies will also shed light on some fundamental questions about the control of cell cycle, cell fate, and their relationship during neural development.

项目概要 无义介导的 mRNA 衰减 (NMD) 关键因子的突变，包括 Upf2、Upf3a、Upf3b 和 Smg6， 富含多种神经发育疾病。除了通过降级来确保转录质量之外 具有过早终止密码子的异常转录本，NMD 调节选择性 mRNA 的稳定性以进行微调 转录丰度。 NMD 是否以及如何影响大脑发育仍然是个谜。我们的长期 目的是了解 NMD 调节在复杂且动态的过程中的功能作用 神经发生及其失调如何导致神经发育障碍。我们确定 通过Upf2的选择性基因消融和体内NMD对神经发育的要求 操纵其他 NMD 因素。我们的初步数据表明，神经干中 UPF2 的缺失 祖细胞导致小头畸形。 UPF2 缺失特别影响细胞周期和谱系进展 放射状胶质细胞（RGC）是发育中的新皮质中的主要神经祖细胞。我们将结合切割 边缘分子细胞核基因组学方法、小鼠遗传学和发育神经生物学进行剖析 NMD 调节神经发生的机制。我们提出三个独立且相互关联的目标 研究小头畸形表型背后的可能变量。在目标 1 中，我们将确定单元格 定性和定量地研究 NMD 敲除小鼠 RGC 的循环行为，并揭示其潜在机制 监管机制。在目标 2 中，我们将确定 RGC 的谱系进展以及由此产生的结果 NMD 敲除小鼠每时间单位的神经元输出。通过表征这些分子细胞缺陷，我们 还旨在为 RGC 谱系转变的转录组调控提供机制见解。 NMD 可能 调节细胞命运要么独立于细胞周期控制，要么作为影响细胞周期的结果。在 目标 3，我们将测试这两个假设，并利用我们的结果重新审视细胞之间的关系 周期和细胞命运。成功完成这些研究将为如何选择性地提供基本见解 mRNA 稳定性是哺乳动物大脑中高度调控的皮质神经发生过程的基础。这 拟议的研究还将阐明有关细胞周期、细胞命运、 以及它们在神经发育过程中的关系。