Cell type signaling specificity of the neurodevelopmental disease-associated DYRK1A kinase

神经发育疾病相关 DYRK1A 激酶的细胞类型信号传导特异性

• 批准号: 10522818
• 负责人: GEORGIA PANAGIOTAKOS
• 金额: $ 42.02万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-08 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10522818
• 关键词: Adoption; Affect; Astrocytes; Automobile Driving; Binding; Brain; Calcineurin; Calcium; Calcium Channel; Calcium Signaling; Cell Differentiation process; Cell Maintenance; Cells; Cerebral cortex; Cues; DNA Binding; Data; Development; Disease; Dose; Down Syndrome; Electroporation; Embryo; Equilibrium; Etiology; Event; Foundations; Functional disorder; Generations; Genes; Genetic; Genetic Transcription; Genetic study; Goals; Human; Human Genetics; Impairment; Intellectual functioning disability; Knowledge; Link; Mental disorders; Microcephaly; Molecular; Morphology; Mouse Strains; Mus; Mutate; Mutation; NFAT Pathway; Neurodevelopmental Disorder; Neuroglia; Neuronal Differentiation; Neurons; Neurosciences; Patients; Phenocopy; Phenotype; Phosphotransferases; Play; Production; Publishing; Radial; Regulation; Research; Role; STAT3 gene; Series; Signal Pathway; Signal Transduction; Specificity; Syndrome; Testing; Therapeutic; Thinness; Timothy syndrome; Transgenic Organisms; autism spectrum disorder; cell behavior; cell type; developmental disease; dosage; excitatory neuron; extracellular; in utero; in vivo; insight; loss of function; loss of function mutation; member; nerve stem cell; nervous system development; neuron apoptosis; neuron loss; neuropsychiatric disorder; neuropsychiatry; neuroregulation; novel; postnatal; programs; relating to nervous system; stem; stem cells; tool; transcription factor

PROJECT SUMMARY/ABSTRACT Mutations in DYRK1A, which encodes a ubiquitously expressed kinase that antagonizes the calcium- dependent calcineurin (CaN)/NFAT signaling pathway, have been reproducibly linked to neurodevelopmental disease. DYRK1A loss of function has been associated with syndromic intellectual disability and autism spectrum disorders (ASD), and increased DYRK1A activity is thought to underlie aspects of Down Syndrome pathophysiology. These genetic clues underscore DYRK1A dosage-dependent regulation of nervous system development; however, the precise mechanisms by which DYRK1A executes its roles in the developing brain remain poorly understood. Our long-term goal is to understand how DYRK1A acts in specific cell types of the embryonic cerebral cortex to influence the commitment of neural stem and progenitor cells to specific neural fates. In the proposed studies, we focus on NFAT-dependent transcriptional mechanisms as primary effectors of DYRK1A activity in neural stem cells and their progeny. We have found that deleting Dyrk1a specifically in the developing cortex differentially impacts calcium signaling in neural stem/progenitor cells and neurons of both the mouse and human. Loss of one or both copies of Dyrk1a results in dose-dependent cortical thinning, depletion of radial glia stem cells, reduced astrocyte abundance, neuronal cell death, and shifts in excitatory neuron differentiation. Our previous studies uncovered similar changes in the generation of excitatory neuron subtypes resulting from the mutation in the Cav1.2 calcium channel that gives rise to the syndromic ASD Timothy Syndrome. Imbalances in these same excitatory neuron types have also been linked to neuropsychiatric syndromes and channelopathies, hinting that calcium-regulated molecular mechanisms may represent a core substrate underlying cellular phenotypes in ASD and other neurodevelopmental disorders. In line with this idea, we have found that the effects of Dyrk1a deletion during cortical development are phenocopied by in vivo modulation of CaN/NFAT signaling, and we have used CUT&RUN sequencing to begin to identify NFAT transcriptional targets in the developing brain. Building on these strong published and preliminary findings, the central objective of this proposal is to define cell type-specific mechanisms by which DYRK1A regulates the development of the cortex. The proposed research tests the ideas that NFAT transcriptional targets underlie deficits in stem cell maintenance and differentiation resulting from cortex-specific Dyrk1a inactivation (AIM 1), that DYRK1A and calcium signaling through CaN/NFAT play key roles in cortical astrogliogenesis (AIM 2), and that cell type-specific NFAT targets contribute to DYRK1A signaling specificity (AIM 3). These studies will build a foundation for future research expanding our knowledge of how calcium signaling regulates brain development and how ubiquitously expressed disease-relevant genes exert specific functions in different cell types. Our results will also provide therapeutic entry points for convergent intracellular mechanisms driving neurodevelopmental disorders.

项目总结/摘要 DYRK 1A编码一种普遍表达的激酶，该激酶拮抗钙离子通道， 依赖性钙调神经磷酸酶（CaN）/NFAT信号通路，已被重复地与神经发育 疾病DYRK 1A功能丧失与综合征性智力残疾和自闭症有关 谱系障碍（ASD）和DYRK 1A活性增加被认为是唐氏综合征的基础 病理生理学这些遗传线索强调了DYRK 1A对神经系统的剂量依赖性调节 然而，DYRK 1A在发育中的大脑中发挥作用的确切机制 仍然知之甚少。我们的长期目标是了解DYRK 1A如何在特定的细胞类型中起作用， 胚胎大脑皮层影响神经干细胞和祖细胞向特定神经细胞的定向分化 命运在所提出的研究中，我们将重点放在NFAT依赖的转录机制作为主要效应子上 DYRK 1A在神经干细胞及其后代中的活性。 我们已经发现，在发育中的皮层中特异性地删除Dyrk 1a对钙的影响是不同的。 在小鼠和人的神经干/祖细胞和神经元中的信号传导。一方或双方的损失 Dyrk 1a的拷贝导致剂量依赖性皮质变薄，放射状胶质干细胞的耗竭， 星形胶质细胞丰度、神经元细胞死亡和兴奋性神经元分化的变化。我们以前的研究 在兴奋性神经元亚型的产生中发现了类似的变化，这些变化是由基因突变引起的。 Cav1.2钙通道，引起ASD综合征蒂莫西综合征。不平衡在这些相同的 兴奋性神经元类型也与神经精神综合征和通道病有关，暗示 钙调节的分子机制可能代表了细胞表型的核心底物， ASD和其他神经发育障碍。根据这一想法，我们发现Dyrk 1a的作用 在皮质发育过程中的缺失是通过体内CaN/NFAT信号转导的调节来表型复制的，我们 已经使用CUT&RUN测序来开始鉴定发育中的大脑中的NFAT转录靶点。 在这些强有力的公布和初步调查结果的基础上，本提案的中心目标是确定 细胞类型特异性机制，DYRK 1A通过该机制调节皮质的发育。拟议 研究测试了NFAT转录靶点是干细胞维持缺陷的基础的想法， 由于皮质特异性Dyrk 1a失活（AIM 1）导致的分化，DYRK 1A和钙信号传导 通过CaN/NFAT在皮质星形胶质细胞发生（AIM 2）中起关键作用，并且细胞类型特异性NFAT靶向 有助于DYRK 1A信号传导特异性（AIM 3）。这些研究将为今后的研究奠定基础 扩展了我们对钙信号如何调节大脑发育以及钙信号如何无处不在的 表达的疾病相关基因在不同的细胞类型中发挥特定的功能。我们的研究结果还将提供 驱动神经发育障碍的会聚细胞内机制的治疗切入点。