Investigating the synaptic pathology of Autism

研究自闭症的突触病理学

• 批准号: 10053341
• 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/10053341
• 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; 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.

项目摘要 赋予自闭症风险的基因突变通常在谷氨酸突触表达的基因中发生。 这些基因的蛋白质产物形成高度相互连接的蛋白质相互作用网络（PIN），并且 代表有吸引力的治疗靶标，因为它们在整个寿命中都表达，并且可以敏锐 用小分子药物靶向。但是，该引脚在正常或 疾病状态知之甚少。在这里，我们应用了一种新颖的针映射技术，定量多路复用 共免疫沉淀，以探索自闭症与谷氨酸的自闭症销钉的投入输出关系 突触响应生理输入。我们的目标系统是一个20-Meb的销钉，由谷氨酸组成 受体，支架和信号转导分子；编码所有靶蛋白的基因中的突变 已经与自闭症有关。我们首先表明，在野生型动物中，我们的目标引脚改变了 以KCL或谷氨酸刺激的急性刺激，以定型方式进行共同交往的模式， 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，以及 证明PIN会产生特定的，可识别的特征，以响应通过 mglur或NMDA受体。在生理谷氨酸刺激的背景下，PIN整合了两者 输入以产生协调的细胞反应增强或去训练。基于这些和其他 初步观察和已发布的数据，我们建议有助于自闭症风险中断的突变 信息流过此引脚，从而使LTP样增强和LTD样之间的平衡 矿化症发生了变化，最终导致了激发和抑制之间的生物水平的失衡。 我们将通过在三个不同的，三个不同的， 自闭症的特征良好的动物模型 - 脆弱的X淘汰赛，Shank3淘汰赛和UBE3A 过表达的模型。我们将表征MGLUR或NMDA刺激的输入输出关系， 并使用主组件空间中的矢量转换模型对其集成进行数学建模。 我们将定义特定机制，通过这些机制，自闭症相关的突变破坏了输入输出关系， 或在生理刺激的背景下破坏信号积分。此外，我们将对待我们的两个 动物模型（shank3和脆弱的X）和先前已证明可以挽救自闭症的药物 - 喜欢行为。我们将在有或没有并发刺激的情况下对销的反应建模 定义与行为救援相关的针签名。总而言之，我们建议（1）定义正常 信息流过由自闭症基因的蛋白质产物组成的销； （2）定义如何 自闭症鼠标模型中的信息流被破坏，目的是充分理解系统 设计有针对性的药物治疗和（3）定义PIN对正确行为的药物的反应， 可以用作设计PIN修改处理的模板，以恢复正常的突触功能。