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 导致自闭症风险的基因突变通常发生在构成信号转导网络的基因中 连接突触传递和下游基因表达的变化。但是，动态的、网络的- 在正常或疾病状态下，这些复杂且相互关联的信号网络的规模行为很差， 明白这是第一次更新申请的一个高生产率的R 01赠款，建立了一个定量多路复用 免疫共沉淀或QMI面板，以研究20个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 编码所有靶蛋白的基因都与自闭症有关。使用QMI，我们发现这个PIN 通过改变协调相互作用模块的组成和强度来编码信息， 接收信号此外，我们发现导致自闭症风险的突变会破坏突触PIN 通过使它们呈现类似于野生型神经元的状态的网络状态， 经历了自我平衡的缩放。这导致网络响应变化的动态范围减小 随后的刺激，并导致系统水平的干扰基础谷氨酸张力（反映在中断 E/I余额）。在第二个周期中，我们关注的问题是，我们能否使ASD PIN正常化， 正常化与表型的功能拯救相关吗？在目标1中，我们重点关注通过FYN进行规范化 激酶，我们将其鉴定为FMR 1-/y小鼠突触可塑性下游的失调网络枢纽 输入。初步数据表明，抑制过度活跃的FYN信号正常化过度活跃的蛋白质 合成和行为;我们建议进行一系列广泛的分子、细胞和行为分析 研究FYN抑制治疗FMR 1缺陷表型的潜力。在目标2中，我们扩展了 对突触可塑性至关重要的第二个PIN，mTOR网络进行网络规模分析。我们使用 药理学和遗传学抑制mTOR，以模拟信息流通过mTOR PIN， 突触可塑性，并建立哪些组成部分的途径所需的稳态缩放，在 体内或体外。在目标3中，我们专注于通过操纵突触活动来实现PIN正常化。我们试图"联合国- 测量携带ASD突变的小鼠的突触PIN，并测量这种治疗是否能够恢复正常 PIN活性和神经元进行正常稳态缩放的能力。关键的是，最后一个实验 将揭示发育机制下游神经元活动水平的改变是否会导致发育机制的中断， 突触和mTOR信号转导，或者相反，如果信号转导中的持续缺陷是独立的， 甚至是基础活动水平改变的原因。总的来说，这一更新将继续我们对 ASD风险基因破坏突触信号转导的分子网络机制。