The role of the mRNA decay factor CNOT3 in cortical development

mRNA衰减因子CNOT3在皮质发育中的作用

• 批准号: 10388793
• 负责人: Lucas Serdar
• 金额: $ 6.76万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-10 至 2025-02-09
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10388793
• 关键词: Address; Affect; Apoptosis; Area; Brain; Cell Cycle; Cell Cycle Regulation; Cell physiology; Cells; Cerebral cortex; Data; Data Set; Defect; Development; Developmental Delay Disorders; Disease; Electroporation; Embryo; Equilibrium; Event; Excision; Exhibits; Frequencies; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Image; Impairment; In Vitro; Inherited; Intellectual functioning disability; Knockout Mice; Knowledge; Label; Lead; Learning Disabilities; Light; Link; Maintenance; Measures; Mediating; Messenger RNA; Metabolic; Microcephaly; Missense Mutation; Monitor; Mus; Mutate; Neurons; Nonsense Mutation; Patients; Poly(A) Tail; Population; Post-Transcriptional Regulation; Process; Proliferating; Proteins; RNA; RNA Decay; RNA Stability; RNA analysis; Radial; Regulation; Role; Technology; Testing; Thick; Transcript; Transcriptional Regulation; Work; autism spectrum disorder; cell type; cognitive function; cohort; conditional knockout; density; excitatory neuron; experimental study; imaging approach; in utero; in vivo; insight; loss of function; mRNA Decay; mRNA Stability; mRNA Transcript Degradation; mouse model; mutant; nerve stem cell; neurogenesis; novel; progenitor; relating to nervous system; self-renewal; stem cell fate; stem cells; transcriptome sequencing

ABSTRACT Proper spatial and temporal control of gene expression during development of the cerebral cortex is critical for the maintenance of neural progenitors and for neural differentiation. While transcriptional regulation has long been recognized as critical to the control of gene expression during cortical development, the contribution of RNA decay is less well understood. Despite this, mis-regulation regulation of gene expression at the post-transcriptional level is emerging as a major cause of neurodevelopmental diseases, including intellectual disability and autism. CNOT3, a regulator of mRNA stability, has recently been found to be mutated in a cohort of patients suffering from intellectual disability and autism. Additional studies have shown that CNOT3 regulates the stability of transcripts encoding proteins involved in cell cycle regulation and stem cell fate. Despite the links between Cnot3, mRNA degradation, neurodevelopmental disease, how Cnot3 contributes to brain development, and its key targets in this process, are unknown. I will address this knowledge gap using a Cre-lox conditional knockout mouse model to ablate Cnot3 expression in neural progenitors. Preliminary data indicates that at embryonic day (E)14.5 (approximately mid-neurogenesis), cKO mice exhibit microcephaly, apoptosis, and a reduction in the number of actively proliferating cells. In this proposal, I will build upon these observations and test the overall hypothesis that that CNOT3-mediated regulation of mRNA stability is required for proper cortical development. Aim 1 investigates the impact of Cnot3 disruption across corticogenesis, and uses live imaging approaches to monitor the requirements for CNOT3 in progenitor fate decisions. Aim 2 uses state-of-the-art technology to globally monitor mRNA stability in neural progenitors and assess the consequences of Cnot3 deletion on mRNA stability. These approaches will reveal previously uncharacterized role for CNOT3 in cortical development, while providing an unprecedented view of RNA stability during cortical development, and how its mis-regulation contributes to disease.

抽象的 大脑皮层发育过程中基因表达的适当空间和时间控制是 对于神经祖细胞的维持和神经分化至关重要。在转录调控的同时 长期以来一直被认为对于皮质发育过程中基因表达的控制至关重要 RNA 衰变的贡献尚不清楚。尽管如此，基因表达的错误调节 转录后水平正在成为神经发育疾病（包括智力障碍）的主要原因 残疾和自闭症。 CNOT3 是 mRNA 稳定性的调节因子，最近在一个队列中被发现发生突变 患有智力障碍和自闭症的患者。其他研究表明 CNOT3 调节 编码参与细胞周期调节和干细胞命运的蛋白质的转录物的稳定性。尽管有链接 Cnot3、mRNA 降解、神经发育疾病、Cnot3 如何促进大脑发育、 及其在此过程中的关键目标尚不清楚。我将使用 Cre-lox 条件来解决这个知识差距 敲除小鼠模型以消除神经祖细胞中的 Cnot3 表达。初步数据表明，在 胚胎日 (E)14.5（大约神经发生中期），cKO 小鼠表现出小头畸形、细胞凋亡和 活跃增殖细胞数量减少。在本提案中，我将基于这些观察和 检验总体假设，即 CNOT3 介导的 mRNA 稳定性调节是正常皮质所必需的。 发展。目标 1 研究 Cnot3 破坏对皮质生成的影响，并使用实时成像 监测祖细胞命运决定中 CNOT3 需求的方法。目标 2 使用最先进的技术 全局监测神经祖细胞中 mRNA 稳定性并评估 Cnot3 后果的技术 缺失对 mRNA 稳定性的影响。这些方法将揭示 CNOT3 在皮质中以前未表征的作用 的发展，同时提供了皮质发育过程中 RNA 稳定性的前所未有的观点，以及它如何 调节不当会导致疾病。