Regulation of the DYRK1A kinase by the Down Syndrome Cell Adhesion Molecule DSCAM

唐氏综合症细胞粘附分子 DSCAM 对 DYRK1A 激酶的调节

• 批准号: 10573072
• 负责人: PETER Gerard FUERST
• 金额: $ 15万
• 依托单位: UNIVERSITY OF IDAHO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-08 至 2023-02-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10573072
• 关键词: Address; Affect; Aneuploidy; Area; Axon; Biochemical; Biological Assay; Blindness; Brain; Brain region; Cell Adhesion Molecules; Cell Death; Cell Nucleus; Cell Surface Receptors; Cell surface; Cellular biology; Cerebellum; Chromosome 16; Chromosome 21; Clinical; Cues; Cytoplasmic Tail; Data; Defect; Dendrites; Development; Disease; Down Syndrome; Down Syndrome Cell Adhesion Molecule; Environment; Event; Eye; Foundations; Future; Gene Duplication; Gene Expression; Genes; Genetic; Genetic study; Goals; Hippocampus (Brain); Human Chromosomes; Idaho; In Vitro; Intervention; Knowledge; Link; Mediating; Modeling; Molecular; Morphology; Mosaicism; Mus; Mutation; Nervous system structure; Neurites; Neurons; Neurophysiology - biologic function; Nuclear; Nuclear Translocation; Organ; Outcome; Pathology; Persons; Phenocopy; Phenotype; Phosphotransferases; Pilot Projects; Play; Population; Proteins; Publications; Reagent; Regulation; Reporting; Research; Research Proposals; Retina; Role; Schizophrenia; Scientist; Signal Pathway; Signal Transduction; Signaling Molecule; Signs and Symptoms; Specificity; Surface; Syndrome; System; Testing; Time; Tissues; Tyrosine Phosphorylation; Universities; Work; autism spectrum disorder; base; cell type; dosage; experience; experimental study; gain of function; human disease; loss of function; mouse Ts65Dn; mouse model; neurodevelopment; novel; overexpression; postnatal; prevent; receptor; relating to nervous system; response; retinal neuron

ABSTRACT: Candidate and Environment: The Fuerst Lab will conduct genetic, molecular and morphological experiments related to the Aims of this proposal at the University of Idaho. The Fuerst lab team has extensive expertise centered on the cell biology and genetics of neural development. Fuerst identified Dscams as the first molecules that regulate mosaic organization of neurons and developed many of the genetic reagents for use in these experiments. Here we will build on our foundation of experience studying Dscam genes to probe the function of downstream signaling networks in neural development. Research Proposal: Developing neurons utilize a range of molecular cues to organize into neural tissues and organs. We and others have previously reported that Down syndrome cell adhesion molecule (Dscam) genes are required for normal development of neural tissues. How Dscam genes mediate a wide range of cellular responses even within a single cell type is an active area of research. In our preliminary studies we found that overexpression of Dyrk1a, whose product interacts biochemically with DSCAM, phenocopies Dscam loss of function. Further we report a redistribution of cellular DYRK1A protein in the Dscam loss of function retina and brain. Based on these observations and the genes’ known antagonistic roles in either promoting or inhibiting developmental cell death, we hypothesize that DSCAM interactions on the cell surface antagonize DYRK1A activity by controlling its subcellular localization. We test this hypothesis in two specific Aims. Aim 1: Pilot genetic studies indicate that either decreasing Dscam dosage or increasing Dyrk1a dosage results in mistargeting of retinal neuron neurites. A similar increase in Dyrk1a expression in the Ts65Dn trisomic mouse models results in a wild type targeting phenotype, however, suggesting a factor or factors in the Down syndrome critical region (DSCR) are compensating for Dyrk1a overexpression. First, we will test if increasing Dyrk1a expression in the Ts65Dn mouse model above the expression of other DSCR genes is sufficient to restore Dyrk1a overexpression neurite targeting defects (Aim 1A). Second, we will test if reducing Dscam expression in Ts65Dn mice to wild type levels restores the Dyrk1a overexpression phenotype (Aim 1B). Aim 2: We will extend our pilot study generated in retina into the brain at large. We will test if DYRK1A localization is developmentally dynamic in the brain (Aim 2A), if DSCAM regulates DYRK1A localization in the brain (Aim 2B) and if Dscam expression is sufficient to modify DYRK1A accumulation or localization in vitro (Aim 2C). Long-term goals: Our long-term goal is to understand how events at the cell surface are transduced to generate different cellular responses. This R03 will facilitate completion and publication of a pilot study, the results of which will inform experiments to understand how regulation of DYRK1A localization impacts its function in development and disease. Significance: Understanding why mutation or overexpression of genes involved in neural development can present with a variety of signs and symptoms in people will require us to also understand the functions of the affected genes with the long-term goal of developing clinical interventions. Here we explore the interaction between two interacting genes in the Down Syndrome critical region, relevant to studies and treatments focusing on either single gene. Understanding the downstream signaling pathways utilized by surface receptors will be essential to help scientists and clinicians pinpoint strategies to treat human diseases.

摘要：候选者和环境：Fuerst 实验室将进行遗传、分子和形态学研究 爱达荷大学与该提案的目标相关的实验。 Fuerst 实验室团队拥有广泛的 专业知识集中于细胞生物学和神经发育遗传学。 Fuerst 将 Dscams 确定为第一个 调节神经元嵌合组织的分子，并开发了许多用于 这些实验。在这里，我们将在研究 Dscam 基因的经验基础上探索 下游信号网络在神经发育中的功能。研究计划：发育神经元 利用一系列分子线索组织成神经组织和器官。我们和其他人之前已经 据报道，唐氏综合症细胞粘附分子（Dscam）基因是正常发育所必需的 神经组织。 Dscam 基因如何介导广泛的细胞反应，甚至在单一细胞类型内也是如此 一个活跃的研究领域。在我们的初步研究中，我们发现 Dyrk1a 的过度表达，其产物 与 DSCAM 发生生化相互作用，表型复制 Dscam 功能丧失。此外，我们报告了重新分配 Dscam 中的细胞 DYRK1A 蛋白丧失视网膜和大脑的功能。基于这些观察和 我们假设基因在促进或抑制发育细胞死亡方面已知的拮抗作用 DSCAM 在细胞表面的相互作用通过控制其亚细胞来拮抗 DYRK1A 活性 本土化。我们在两个具体目标中测试这个假设。目标 1：试点遗传学研究表明 减少 Dscam 剂量或增加 Dyrk1a 剂量会导致视网膜神经元突起的定位错误。一个 Ts65Dn 三体小鼠模型中 Dyrk1a 表达的类似增加导致野生型靶向 然而，表型表明唐氏综合症关键区 (DSCR) 中的一个或多个因素是 补偿 Dyrk1a 过度表达。首先，我们将测试是否增加 Ts65Dn 中的 Dyrk1a 表达 高于其他 DSCR 基因表达的小鼠模型足以恢复 Dyrk1a 过度表达 神经突靶向缺陷（目标 1A）。其次，我们将测试是否将 Ts65Dn 小鼠中的 Dscam 表达降低至野生状态 类型水平恢复 Dyrk1a 过度表达表型（目标 1B）。目标 2：我们将扩大试点研究 在视网膜中产生并进入大脑。我们将测试 DYRK1A 本地化是否具有发展活力 大脑（目标 2A），如果 DSCAM 调节 DYRK1A 在大脑中的定位（目标 2B）并且如果 Dscam 表达是 足以改变 DYRK1A 体外积累或定位（目标 2C）。 长期目标：我们的长期目标是了解细胞表面的事件如何转导为 产生不同的细胞反应。该 R03 将促进试点研究的完成和发布，该研究 其结果将为实验提供信息，以了解 DYRK1A 定位的调控如何影响其 在发育和疾病中发挥作用。 意义：了解为何神经发育中涉及的基因发生突变或过度表达 人类可能会出现各种体征和症状，因此我们还需要了解其功能 受影响的基因，长期目标是开发临床干预措施。下面我们就来探讨一下交互 唐氏综合症关键区域中两个相互作用基因之间的关系，与研究和治疗相关 专注于任一单个基因。了解表面利用的下游信号通路 受体对于帮助科学家和临床医生确定治疗人类疾病的策略至关重要。