Defining links between an intellectual disability associated RNA-binding protein and planar cell polarity in neurodevelopment

定义智力障碍相关的 RNA 结合蛋白与神经发育中平面细胞极性之间的联系

• 批准号: 9761046
• 负责人: Edwin Corgiat
• 金额: $ 4.5万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9761046
• 关键词: Affect; Afferent Neurons; Alleles; Axon; Biological; Biological Assay; Biological Process; Brain; Cells; Cochlea; Cryoultramicrotomy; Cytoplasmic Granules; Data; Data Set; Defect; Dendrites; Dependence; Development; Distal; Doctor of Philosophy; Dorsal; Dose; Drosophila genus; Drosophila melanogaster; Etiology; Exhibits; Functional disorder; Gene Expression; Genes; Genetic; Genetic Techniques; Genome; Goals; Hand; Heterogeneity; Human; Impairment; Individual; Inherited; Intellectual functioning disability; Knowledge; Larva; Lead; Learning; Length; Life; Link; Locomotion; Memory; Messenger RNA; Metabolism; Microtubules; Modeling; Molecular; Movement; Mushroom Bodies; Mutant Strains Mice; Mutation; Nerve Tissue; Nervous system structure; Neurites; Neurons; Neuropil; Nuclear; Olfactory Learning; Orthologous Gene; Pathway Analysis; Pathway interactions; Patients; Phenotype; Play; Poly(A) Tail; Poly(A)+ RNA; Population; Post-Transcriptional Regulation; Process; Protein Biosynthesis; Proteins; Proteome; Proteomics; Pupa; RNA; RNA Interference; RNA-Binding Proteins; Reading; Role; Signal Transduction; Sterile coverings; Structure; System; Techniques; Testing; Tissues; Training; Transgenes; Transgenic Organisms; Translations; Twin Multiple Birth; WNT Signaling Pathway; Work; Zinc Fingers; axon guidance; axonal guidance; base; cell type; cognitive function; cost; fly; genetic approach; in vivo; insight; link protein; man; messenger ribonucleoprotein; mutant; nervous system development; neurodevelopment; neuron loss; neuronal survival; planar cell polarity; polyadenosine; relating to nervous system; tool; training opportunity

PROJECT SUMMARY: Intellectual disability affects 1–3% of the worldwide population; these individuals have deficits in adaptive functioning necessitating ongoing support to perform activities such as dressing, reading, and interpreting the intentions of others. The cost of this support, for a single US patient, is approximately $1–2 million throughout his or her lifespan1. The pathophysiology and etiology of intellectual disability has been difficult to elucidate due to the heterogeneity in casual mutations2. Despite this complexity, mutations linked to intellectual disability tend to accumulate in pathways relating to nervous system development, cellular metabolism, and microtubule based movement and axonal transport3. Additionally, monogenic forms of intellectual disability provide direct insight into defective cell biological processes that underlie intellectual disability. Our lab found that mutations in a ubiquitously expressed zinc-finger, polyadenosine RNA-binding protein, ZC3H14, are linked to a form of monogenic, non-syndromic autosomal recessive intellectual disability4. We developed a Drosophila melanogaster model to investigate the role of dNab2, the fly ortholog to human ZC3H14. Loss of dNab2 results in neuronal, survival, and locomotive phenotypes. Importantly, many of the phenotypes can be rescued by transgenic expression of human ZC3H14 exclusively in neurons, implying a conservation of function from flies to humans and feasibility in using dNab2 to model ZC3H14 function. Data generated from our fly model suggests that dNab2 loss is critical in neurodevelopment, and that it may be regulating gene expression in neurons. However, the identity of mRNA targets of dNab2 in neurons and the mechanism by which it regulates these targets are key gaps in knowledge. Preliminary data suggest that dNab2 interacts functionally with multiple components of the planar cell polarity (PCP) pathway. PCP is a non-canonical branch of Wnt signaling that regulates axon guidance in the nervous system and tissue polarization in somatic tissue 5–8. Therefore, I will directly test the hypothesis that dNab2 regulates PCP components to control neurite extension and guidance during neurodevelopment. The Specific Aims of this project are: 1) define genetic dNab2:PCP interactions in two neurodevelopmental contexts, 2) utilize a systems-level proteomic approach to identify dNab2 regulated pathways in the brain, and 3) examine physical and functional interactions between dNab2 and PCP pathway RNAs in vivo. Aim 1 utilizes Drosophila genetic tools to assess functional genetic interactions with dNab2 in two neuronal cell types. Aim 2 uses network analyses, of a unique Drosophila proteomic dataset, to identify dNab2 regulated pathways. Aim 3 utilizes techniques to assess RNA localization and physical interaction with dNab2. Successful completion of these aims will provide insight into how dNab2 regulates local gene expression to impact neurodevelopment, and thus support our broad, long-term objective of defining the cell biological and molecular mechanisms underlying ZC3H14/dNab2 related neurodevelopmental defects.

项目总结： 智力残疾影响着世界人口的1-3%；这些人在适应能力方面存在缺陷 需要持续的支持来执行诸如穿衣、阅读和口译等活动 其他人的意图。对于一名美国患者来说，这种支持的成本在整个过程中大约为100-200万美元 他或她的一生1。智力残疾的病理生理学和病因学一直难以阐明。 偶然突变中的异质性2。尽管这种复杂性，与智力残疾有关的突变往往 在与神经系统发育、细胞代谢和微管相关的途径中积累 运动和轴突运输。此外，单基因形式的智力残疾提供了直接的洞察力 转化为有缺陷的细胞生物学过程，这是智力残疾的基础。我们的实验室发现，一种 普遍表达的锌指多聚腺苷RNA结合蛋白ZC3H14与一种形式的 单基因、非综合征型常染色体隐性智力残疾。我们培育了一只果蝇 研究与人类ZC3H14同源的果蝇dNab2基因在黑胃动物模型中的作用。丢失dNab2结果 在神经元、存活和运动表型上。重要的是，许多表型可以通过 人ZC3H14的转基因表达仅在神经元中，暗示果蝇的功能保守 以及用dNab2模拟ZC3H14函数的可行性。从我们的苍蝇模型产生的数据表明 DNab2的缺失对神经发育至关重要，它可能正在调节神经元中的基因表达。 然而，神经元中dNab2的mRNA靶标的同一性及其调节这些靶标的机制 目标是知识中的关键差距。初步数据表明，dNab2与多个 平面细胞极性(PCP)途径的组成部分。PCP是WNT的一个非规范分支，表示 调节神经系统的轴突引导和躯体组织中的组织极化5-8。因此，我将 直接验证dNab2调控PCP成分控制轴突延伸和引导的假说 在神经发育过程中。这个项目的具体目标是：1)定义基因dNAb2：PCP在 两种神经发育背景，2)利用系统水平的蛋白质组学方法来识别dNab2调节的 大脑中的通路，以及3)检查dNab2和PCP通路之间的物理和功能相互作用 活体内的RNA。目的1利用果蝇遗传工具评估与dNab2在两个区域的功能遗传相互作用 神经细胞类型。Aim 2使用一个独特的果蝇蛋白质组数据集的网络分析来鉴定dNab2 受调控的通路。AIM 3利用技术来评估RNA定位和与dNab2的物理相互作用。 这些目标的成功完成将为深入了解dNab2如何调节局部基因表达提供依据 影响神经发育，从而支持我们定义细胞生物学和 ZC3H14/dNab2相关神经发育缺陷的分子机制