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：候选人与环境：富尔斯特实验室将在2010年10月20日至21日对候选人进行遗传、分子和形态学研究。 在爱达荷州大学进行的与该提案目标相关的实验。富尔斯特实验室团队拥有广泛的 专业知识集中在神经发育的细胞生物学和遗传学。富尔斯特认为Dscams是第一个 这些分子调节神经元的镶嵌组织，并开发了许多遗传试剂用于 这些实验。在这里，我们将建立在我们研究Dscam基因的经验基础上， 下游信号网络在神经发育中的作用。研究方向：神经元发育 利用一系列分子线索组织成神经组织和器官。我们和其他人以前 报道唐氏综合征细胞粘附分子（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，即使在单一细胞类型内， 一个活跃的研究领域在我们的初步研究中，我们发现Dyrk1a的过表达，其产物 与DSCAM发生生物化学相互作用，表型模仿DSCAM功能丧失。此外，我们还报告了 细胞DYRK1A蛋白在Dscam视网膜和大脑功能丧失中的作用。根据这些观察和 基因在促进或抑制发育性细胞死亡中的已知拮抗作用，我们假设 细胞表面的DSCAM相互作用通过控制DYRK1A的亚细胞活性来拮抗DYRK1A的活性， 本地化我们在两个具体目标中检验这一假设。目的1：初步遗传研究表明， 降低Dscam剂量或增加Dyrk1a剂量导致视网膜神经元神经突的错误定位。一 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，表型表明唐氏综合征关键区（DSCR）中的一个或多个因子是 补偿Dyrk1a过表达。首先，我们将测试增加Ts65Dn中的Dyrk1a表达是否 高于其他DSCR基因表达的小鼠模型足以恢复Dyrk1a过表达 神经突靶向缺陷（Aim 1A）。第二，我们将测试是否减少Ts65Dn小鼠中的Dscam表达至野生型， 型水平恢复Dyrk1a过表达表型（Aim 1B）。目标2：我们将扩大试点研究 视网膜产生的放射性物质进入大脑我们将测试DYRK1A本地化是否是动态发展的， 脑（Aim 2A），如果DSCAM调节DYRK1A在脑中的定位（Aim 2B），如果Dscam表达是 足以在体外改变DYRK1A的积累或定位（目的2C）。 长期目标：我们的长期目标是了解细胞表面的事件是如何被转导到 产生不同的细胞反应。这份报告将促进完成和公布一项试点研究， 其结果将为实验提供信息，以了解DYRK1A定位的调节如何影响其功能。 在发育和疾病中发挥作用。 意义：理解为什么参与神经发育的基因突变或过度表达 人们可能会出现各种体征和症状，这也需要我们了解 受影响的基因与发展临床干预的长期目标。在这里，我们探讨的互动 唐氏综合症关键区域中两个相互作用的基因之间的关系，与研究和治疗有关 专注于任何一个基因了解表面活性剂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。