Uncovering the Role of TRIO in Synaptic Function and Autism Spectrum Disorder

项目负责人/主要研究者（最后，第一，中间）：Herring，布鲁斯，E。 项目摘要 自闭症谱系障碍（ASD）是精神损害的主要原因，目前还没有治愈方法。 越来越多的证据表明，突触后突触后神经元突触后能 突触功能作为ASD的基础。我们最近发现了一个前所未有的ASD相关的集群， 突触肌动蛋白调节蛋白Trio的GEF 1结构域中的突变，导致强的全基因组 TRIO基因与ASD的统计学关联。我们研究的长期目标是确定核心突触 调节机制，许多ASD引起因素汇聚在其上。突触识别 ASD风险基因的“汇聚点”将有助于简化这种疾病的遗传景观，从而有助于 开发新的策略来治疗具有多种ASD致病因素的个体。我们的中央 一种假设是，Trio中的ASD突变破坏了大量突触调节途径， 这些途径的结果在突触功能障碍，有助于ASD的发展， 相关的行为表型。在强有力的初步数据的指导下，我们将在三个具体的方面探讨这一假设。 目标。在目标1中，我们将联合收割机结合蛋白质组学、生物化学、电生理学和超分辨率成像 技术，以确定新的突触调控机制，涉及三。在目标2中，我们将联合收割机 用计算机建模的方法来揭示Trio相关的突触调节机制被破坏， 自闭症相关的突变，并提供了一个全面的图片突触中断，结果 自闭症特异性三重功能障碍。而且，对于目标3，我们已经设计了一种条件性基因敲入小鼠， ASD相关突变型Trio的CRE依赖性表达。利用这种新的强大的遗传工具，我们 将进行一系列行为测试，以评估ASD相关的Trio突变对哺乳动物的影响。 行为现在越来越多的证据表明，神经感觉处理缺陷是导致 许多常见的ASD相关行为表型的发展。因此，我们建议使用 最先进的技术，允许仔细检查这些小鼠的体感处理。的 目前的建议是创新的，因为它汇集了一个具有不同专业知识领域的合作者团队， 部署新的和强大的遗传工具，将允许一个多维度的方法来了解如何 突触功能的破坏导致ASD。该提案意义重大，因为它确定了一个重要的 突触信号中枢，连接先前与ASD有关的许多突触蛋白，并将垂直 推进我们对ASD从突触到电路再到行为的理解。这项建议完全符合使命 NINDS的目标是关注突触功能障碍如何最终导致关键的神经功能缺损 这可能是与ASD相关的许多核心行为表型的基础。 OMB编号0925-0001/0002（2018年1月修订版批准至2020年3月31日）续格式页

Program Director/Principal Investigator (Last, First, Middle): Herring, Bruce, E. PROJECT SUMMARY Autism Spectrum Disorder (ASD) is a leading cause of mental impairment for which there is no known cure. Mounting evidence points to a convergence on altered actin-mediated regulation of postsynaptic glutamatergic synaptic function as a basis for ASD. We have recently identified an unprecedented clustering of ASD-related mutations in the GEF1 domain of the synaptic actin regulatory protein, Trio, that results in a strong genome-wide statistical association of the TRIO gene with ASD. The long-term goal of our research is to identify core synaptic regulatory machinery onto which numerous ASD causing factors converge. Identification of synaptic “convergence points” of ASD-risk genes will help simplify the genetic landscape of this disorder and thus, aid in the development of new strategies to treat individuals with a diverse array of ASD-causing factors. Our central hypothesis is that ASD mutations in Trio disrupt a multitude of synaptic regulatory pathways, and that disruption of these pathways results in glutamatergic synapse dysfunction that contributes to the development of ASD- related behavioral phenotypes. Guided by strong preliminary data we will pursue this hypothesis in three specific aims. In Aim 1, we will combine proteomic, biochemical, electrophysiological, and super-resolution imaging techniques to identify novel synaptic regulatory mechanisms involving Trio. In Aim 2, we will combine these same approaches with computational modeling to reveal Trio-related synaptic regulatory mechanisms disrupted by Autism-related mutations and provide a comprehensive picture of the synaptic disruption that results from Autism-specific Trio dysfunction. And, for Aim 3, we have engineered a conditional knock-in mouse that allows CRE-dependent expression of an ASD-related mutant form of Trio. Using this new and powerful genetic tool, we will conduct a battery of behavioral tests to assess the impact of ASD-related Trio mutations on mammalian behavior. Growing evidence now suggests that neurological sensory processing deficits underlie the development of many common ASD-related behavioral phenotypes. Because of this, we propose the use of state-of-the-art techniques that allow careful examination of somatosensory processing in these mice. The present proposal is innovative because it assembles a team of collaborators with diverse areas of expertise and deploys new and powerful genetic tools that will allow a multi-dimensional approach to understanding how disruption of synaptic function leads to ASD. The proposal is significant because it stands to identify an important synaptic signaling hub that links numerous synaptic proteins previously implicated in ASD and will vertically advance our understanding of ASD from synapse to circuit to behavior. This proposal squarely meets the mission objectives of the NINDS given its focus on how synaptic dysfunction ultimately leads to key neurological deficits that likely underlie many core behavioral phenotypes associated with ASD. OMB No. 0925-0001/0002 (Rev. 01/18 Approved Through 03/31/2020) Continuation Format Page