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: 10618819
• 负责人: Lucas Serdar
• 金额: $ 7.18万
• 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/10618819
• 关键词: Ablation; Address; Affect; Apoptosis; Area; Brain; Cell Cycle; Cell Cycle Regulation; Cell Proliferation; 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; Image; Impairment; In Vitro; Inherited; Intellectual functioning disability; Knockout Mice; Knowledge; Label; Learning Disabilities; Link; Maintenance; Measures; Mediating; Messenger RNA; Metabolic; Microcephaly; Missense Mutation; Monitor; Mus; Mutate; Neurodevelopmental Disorder; 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; neural; neurogenesis; novel; posttranscriptional; progenitor; 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 在大脑皮层发育过程中，基因表达的适当空间和时间控制是 维持神经祖细胞和神经分化至关重要。同时转录调节 长期以来，人们一直被认为是控制基因表达在皮质发育过程中的至关重要的关键， RNA衰减的贡献知之甚少。尽管如此，基因表达的错误调节在 转录后级别正在成为神经发育疾病的主要原因，包括智力 残疾和自闭症。 CNOT3是mRNA稳定性的调节剂，最近已被发现在队列中被突变 患有智障和自闭症的患者。其他研究表明，CNOT3调节 编码涉及细胞周期调节和干细胞命运的蛋白质的转录本的稳定性。尽管有链接 在CNOT3，mRNA降解，神经发育疾病之间，CNOT3如何促进大脑发育， 并且其在此过程中的主要目标是未知的。我将使用CRE-LOX条件解决此知识差距 敲除小鼠模型在神经祖细胞中消除CNOT3表达。初步数据表明在 胚胎日（E）14.5（大约在神经发生），CKO小鼠表现出小头畸形，凋亡和A 积极增殖细胞的数量减少。在此提案中，我将以这些观察为基础 测试总体假设，即需要正确皮质的CNOT3介导的mRNA稳定性调节 发展。 AIM 1研究CNOT3跨皮质发生的影响，并使用实时成像 在祖细胞命运决策中监视CNOT3要求的方法。 AIM 2使用最先进的 全球监测神经祖细胞中mRNA稳定性的技术并评估CNOT3的后果 mRNA稳定性上的删除。这些方法将揭示CNOT3先前未表征的角色 开发，同时为皮质发育过程中的RNA稳定性提供了前所未有的观点，以及如何 错误调节会导致疾病。