Investigating the synaptic pathology of Autism

研究自闭症的突触病理学

• 批准号: 10582939
• 负责人: Stephen Edward Paucha Smith
• 金额: $ 79.3万
• 依托单位: SEATTLE CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-01 至 2027-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10582939
• 关键词: Affect; Agonist; Autopsy; Award; Bathing; Behavior; Behavioral Assay; Biological; Biology; Cellular Assay; Characteristics; Co-Immunoprecipitations; Complex; DNA Sequence Alteration; Data; Development; Disease; Electroencephalography; Electrophysiology (science); Equilibrium; Event; FMR1; FRAP1 gene; FYN gene; Feedback; Gene Expression; Gene Mutation; Genes; Genetic; Genome; Glutamate Receptor; Glutamates; Grant; Homeostasis; Human; Hyperactivity; In Vitro; Investigation; Link; Measures; Mediating; Membrane; Modeling; Molecular; Mus; Mutation; Neurodevelopmental Disorder; Neurons; PI3K/AKT; Pathology; Pathway interactions; Patients; Pattern; Phase; Phenotype; Phosphotransferases; Productivity; Protein Biosynthesis; Protein Dynamics; Proteins; Risk; Scaffolding Protein; Signal Transduction; Stimulus; Synapses; Synaptic Transmission; Synaptic plasticity; System; Testing; Vibrissae; autism spectrum disorder; behavioral phenotyping; drug candidate; drug development; experimental study; eyeblink conditioning; gene product; genetic risk factor; in vivo; information model; information processing; inhibitor; mTOR Inhibitor; mTOR inhibition; mTOR protein; mature animal; member; molecular phenotype; mouse genetics; mouse model; neurotransmission; novel; novel therapeutics; pharmacologic; receptor; response; risk variant; scaffold; stem

PROJECT SUMMARY Genetic mutations that confer autism (ASD) risk often occur in genes that comprise signal transduction networks that link synaptic transmission to downstream changes in gene expression. However, the dynamic, network- scale behavior of these complex and interconnected signaling networks in normal or disease states is poorly understood. This is the first renewal application of a highly productive R01 grant that built a quantitative multiplex co-immunoprecipitation or QMI panel to study the dynamic activity of a 20-member protein interaction network (PIN), consisting of glutamate receptors, scaffolds, and signal transduction molecules; mutations in the genes encoding all target proteins have been genetically linked to autism. Using QMI, we discovered that this PIN encodes information by varying the composition and intensity of modules of coordinated interactions in response to incoming signals. Moreover, we found that mutations that contribute to autism risk disrupt synaptic PINs by causing them to assume a network state that resembles the state of a wildtype neuron that has undergone homeostatic scaling. This results in a reduced dynamic range of the network to change in response to subsequent stimuli, and leads to a systems-level disturbance in basal glutamate tone (as reflected in disrupted E/I balance). In the second cycle we focus on the question of, can we normalize ASD PINs, and will this normalization correlate with functional rescue of phenotypes? In Aim 1, we focus on normalization via FYN kinase, which we identified as a dysregulated network hub in FMR1-/y mice downstream of synaptic plasticity inputs. Preliminary data demonstrate that inhibition of hyperactive FYN signaling normalizes hyperactive protein synthesis and behavior; we propose to perform an extensive battery of molecular, cellular and behavioral assays to investigate the potential of FYN inhibition to treat the phenotypes of FMR1 deficiency. In Aim 2, we extend our network-scale analysis to a second PIN critical to synaptic plasticity, the mTOR network. We use pharmacological and genetic inhibition of mTOR to model information flow through the mTOR PIN during synaptic plasticity, and to establish which components of the pathway are required for homeostatic scaling, in vivo or in vitro. In Aim 3, we focus on PIN normalization by manipulating synaptic activity. We attempt to `un- scale' the synaptic PIN of ASD-mutation-carrying mice, and measure if this treatment is able to restore normal PIN activity and the ability of the neuron to undergo normal homeostatic scaling. Critically, this last experiment will reveal whether altered levels of neuronal activity downstream of developmental mechanisms cause disrupted synaptic and mTOR signal transduction, or conversely if ongoing deficits in signal transduction are independent, or even causative, of altered basal activity levels. Overall, this renewal would continue our investigations into the molecular network mechanisms by which ASD risk genes disrupt synaptic signal transduction.

项目摘要 赋予自闭症（ASD）风险的基因突变经常在构成信号转导网络的基因中发生 该连接突触传播与基因表达的下游变化。但是，动态，网络 - 正常或疾病状态中这些复杂和相互联系的信号网络的比例行为很差 理解。这是高产R01赠款的第一次续签应用，该赠款构建了定量多重 共免疫沉淀或QMI面板研究20成员的蛋白质相互作用网络的动态活性 （PIN），由谷氨酸受体，支架和信号转导分子组成；基因突变 编码所有靶蛋白的编码已与自闭症有关。使用QMI，我们发现这个销钉 通过改变协调相互作用的模块的组成和强度来编码信息以响应 传入信号。此外，我们发现有助于自闭症风险破坏突触引脚的突变 通过使他们采用类似于具有野生型神经元状态的网络状态 经过稳态缩放。这导致网络的动态范围减少以改变响应 随后的刺激，并导致基底谷氨酸酸盐的系统级干扰（如被破坏反映 E/i平衡）。在第二个周期中，我们关注的问题，我们可以将ASD引脚正常化，并且会这样 归一化与表型的功能拯救相关？在AIM 1中，我们专注于通过FYN的归一化 激酶，我们确定为突触可塑性下游的FMR1-/Y小鼠中的网络中心失调 输入。初步数据表明，抑制过度活跃的FYN信号传导使多动蛋白归一化 综合和行为；我们建议执行大量分子，细胞和行为测定 研究FYN抑制作用治疗FMR1缺乏症的表型的潜力。在AIM 2中，我们扩展了我们的 网络尺度分析对突触可塑性至关重要的第二引脚，即mTOR网络。我们使用 MTOR的药理和遗传抑制，以模拟通过MTOR PIN的信息流过MTOR PIN 突触可塑性，并确定稳态缩放需要哪些途径的组成部分 体内或体外。在AIM 3中，我们通过操纵突触活动来关注PIN归一化。我们试图不 缩放'ASD致电小鼠的突触销，并测量该处理是否能够恢复正常 销活动和神经元进行正常稳态缩放的能力。至关重要的是最后一个实验 将揭示发育机制下游的神经元活动水平是否导致破坏 突触和MTOR信号转导，或者相反，如果信号转导的持续缺陷是独立的， 甚至因果关系，基础活性水平改变。总体而言，这种续约将继续我们对 ASD风险基因破坏突触信号转导的分子网络机制。