Investigating the synaptic pathology of Autism

研究自闭症的突触病理学

• 批准号: 10292984
• 负责人: Stephen Edward Paucha Smith
• 金额: $ 54.53万
• 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 2022-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10292984
• 关键词: Acute; Affect; Agonist; Animal Model; Animals; Behavior; Behavioral; Binding; Characteristics; Chemicals; Chemosensitization; Co-Immunoprecipitations; Complex; DNA Sequence Alteration; Data; Disease; Drug Targeting; Equilibrium; Feedback; Fragile X Syndrome; Gene Mutation; Gene Proteins; Genes; Genetic Models; Genetic study; Glutamate Receptor; Glutamates; Goals; Homeostasis; Knock-out; Link; Longevity; Measurement; Metabotropic Glutamate Receptors; Modeling; Molecular; Morphology; Mutation; N-Methyl-D-Aspartate Receptors; N-Methylaspartate; Neurodevelopmental Disorder; Neurons; Organism; Output; Pathogenesis; Pathologic; Pathology; Pattern; Pharmaceutical Preparations; Physiological; Process; Protein Engineering; Proteins; Publishing; Risk; Science; Signal Transduction; Slice; Stereotyping; Structure; Synapses; System; Technology; Testing; Therapeutic; Translating; UBE3A gene; Variant; antagonist; autism spectrum disorder; base; design; gene product; graph theory; in vitro Assay; information processing; insight; kinase inhibitor; link protein; mathematical model; member; mouse model; network models; new technology; novel; overexpression; predictive modeling; protein complex; protein function; response; scaffold; small molecule; stem; synaptic function; targeted treatment; therapeutic target; tool; transmission process; vector

PROJECT SUMMARY Genetic mutations that confer autism risk often occur in genes that are expressed at the glutamate synapse. The protein products of these genes form a highly interconnected protein interaction network (PIN), and represent attractive therapeutic targets since they are expressed throughout the lifespan and can be acutely targeted with small molecule drugs. However, the dynamic, network-scale behavior of this PIN in normal or disease states is poorly understood. Here, we apply a novel PIN-mapping technology, quantitative multiplex co-immunoprecipitation, to explore the input-output relationships of an autism-linked PIN at the glutamate synapse as it responds to physiological inputs. Our target system is a 20-member PIN, consisting of glutamate receptors, scaffolds, and signal transduction molecules; mutations in the genes encoding all target proteins have been genetically linked to autism. We first show that, in wild-type animals, our target PIN changes its pattern of co-associations in a stereotyped manner in response to acute stimulation with KCl or glutamate, using cultured neurons or acute slices. We then model the input-output relationships of the PIN system, and demonstrate that the PIN produces specific, recognizable signatures in response to stimulation through the mGluR or NMDA receptors. In the context of physiological glutamate stimulation, the PIN integrates the two inputs to produce a coordinated cellular response- potentiation or de-potentiation. Based on these and other preliminary observations and published data, we propose that mutations that contribute to autism risk disrupt information flow through this PIN, such that the balance between LTP-like potentiation and LTD-like depotentiaion is altered, ultimately leading to an organism-level imbalance between excitation and inhibition. We will test this hypothesis by modeling the PIN response to mGluR or NMDA stimulation in three distinct, well-characterized animal models of autism- the Fragile X knockout, Shank3 knockout, and Ube3a overexpressing models. We will characterize the input-output relationships for mGluR or NMDA stimulation, and mathematically model their integration using a vector transformation model in principal component space. We will define specific mechanisms by which autism-linked mutations disrupt either input-output relationships, or disrupt signal integration in the context of physiological stimulation. In addition, we will treat two of our animal models (Shank3 and Fragile X) with drugs that have been previously demonstrated to rescue autism- like behaviors. We will model the response of the PIN to the drug with or without concurrent stimulation to define a PIN signature associated with behavioral rescue. In summary we propose to (1) define normal information flow through a PIN consisting of the protein products of autism-liked genes; (2) define how information flow is disrupted in mouse models of autism, with the goal of understanding the system sufficiently to design targeted drug treatments and (3) define how the PIN responds to drugs that correct behavior, which could serve as a template for the design of PIN-modifying treatments to restore normal synaptic function.

项目总结 导致自闭症风险的基因突变通常发生在谷氨酸突触表达的基因中。 这些基因的蛋白质产物形成一个高度互联的蛋白质相互作用网络(PIN)，并且 代表有吸引力的治疗靶点，因为它们在整个生命周期中都有表达，并且可能是尖锐的 以小分子药物为靶标。但是，此PIN的动态网络规模行为在正常或 人们对疾病状态知之甚少。在这里，我们应用了一种新的PIN映射技术--定量多路传输 免疫共沉淀，探索自闭症相关谷氨酸PIN的输入-输出关系 突触对生理输入做出反应。我们的目标系统是一个由20个成员组成的PIN，由谷氨酸组成 受体、支架和信号转导分子；编码所有目标蛋白的基因突变 与自闭症有基因上的联系。我们首先展示了，在野生动物中，我们的目标PIN改变了它的 对氯化钾或谷氨酸的急性刺激以刻板印象的方式相互联系的模式， 使用培养的神经元或急性切片。然后，我们对PIN系统的输入输出关系进行建模，并 证明PIN产生特定的、可识别的签名，以响应通过 MGluR或NMDA受体。在生理谷氨酸刺激的背景下，PIN将两者结合在一起 产生协调的细胞反应的输入--增强或去增强。基于这些和其他 根据初步观察和已发表的数据，我们认为导致自闭症的突变有中断的风险。 信息流通过此PIN，因此类LTP增强和类LTD之间的平衡 去电位化被改变，最终导致生物体水平的兴奋和抑制之间的失衡。 我们将通过对三种不同的PIN对mGluR或NMDA刺激的反应建模来验证这一假设， 典型的自闭症动物模型--脆性X基因敲除、Shank3基因敲除和Ube3a基因敲除 过度表达模特。我们将描述mGluR或NMDA刺激的输入-输出关系， 并使用主成分空间中的矢量变换模型对它们的集成进行数学建模。 我们将定义与自闭症相关的突变破坏输入-输出关系的具体机制， 或在生理刺激的背景下扰乱信号整合。此外，我们将治疗我们的两名 动物模型(Shank3和Fragile X)使用先前已被证明可以拯救自闭症的药物- 类似的行为。我们将模拟在有或没有同时刺激的情况下PIN对药物的反应 定义与行为救援关联的PIN签名。综上所述，我们建议(1)定义正常 信息流通过由自闭症样基因的蛋白质产物组成的PIN；(2)定义如何 在自闭症小鼠模型中，信息流被扰乱，目的是充分了解系统 设计有针对性的药物治疗和(3)定义PIN如何对纠正行为的药物做出反应， 可作为设计PIN修饰治疗以恢复正常突触功能的模板。