Elucidating the Roles of SHANK3 and FXR in the Autism Interactome

阐明 SHANK3 和 FXR 在自闭症相互作用组中的作用

• 批准号: 7842983
• 负责人: HUDA Y ZOGHBI
• 金额: $ 40.35万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7842983
• 关键词: 22q13.3; Address; Alleles; Antibodies; Autistic Disorder; Behavioral; Binding; Biochemical; Child; Communities; Complex; Databases; Disease; FMRP; FXR1 gene; FXR2 gene; Foundations; Fragile X Mental Retardation Protein; Fragile X Syndrome; Genes; Genetic; Genomics; Grant; Heterogeneity; Knock-in Mouse; Knockout Mice; Lead; Maintenance; Metabolism; Modeling; Mus; Mutation; Partner in relationship; Pathogenesis; Pathway interactions; Patients; Phenotype; Physiological; Play; Population; Prevalence; Proteins; Public Health; RNA; RNA Processing; Reagent; Recovery; Research; Research Personnel; Resources; Role; Structure; Synapses; Syndrome; TSC1 gene; TSC2 gene; Translations; Tuberous sclerosis protein complex; autism spectrum disorder; base; farnesoid X-activated receptor; human TSC1 protein; in vivo; insight; mouse model; novel; paralogous gene; postsynaptic density protein; protein complex; public health relevance; scaffold; synaptogenesis; therapeutic target

DESCRIPTION (provided by applicant): This proposal is responsive to RFA-MH-09-170, the Recovery Act Limited Competition: Research to Address the Heterogeneity in Autism Spectrum Disorders (R01). As noted in the RFA, Autism spectrum disorders (ASD) form a very heterogeneous group: over 30 distinct genes have been identified as either causing or contributing to the cause of autism. To better understand how such a diverse array of genes can cause similar ASD phenotypes, we searched for the binding partners of proteins encoded by autism-associated genes and developed an autism protein interaction network (an "interactome") that contains over 900 protein interactions, many of which are novel. One of the most exciting discoveries unveiled by this interactome is the finding that there are three "hub" proteins that are centers for interaction: FXR1, SHANK3, and TSC1. Moreover, SHANK3 interacts with two distinct classes of proteins: one involved in synapse structure, the other involved in RNA metabolism and translation. This latter class connects SHANK3 to the Fragile X protein (FMRP) and its paralogs (FXR1 and 2) as well as the tuberous sclerosis complex proteins (TSC1 and TSC2). Based in part on these findings, we hypothesize that there are shared pathogenic mechanisms amongst various ASD, one of which centers on synapse maintenance and plasticity. We propose that SHANK3 plays two distinct roles at the synapse, one involving synapse organization and maintenance and the other involving local RNA translation at the synapse. To gain insight into key mechanisms underlying ASD pathogenesis, we will identify and characterize the native SHANK3 complexes at the synapse in vivo. To accomplish this, we will generate knock-in mice that tag the SHANK3 protein to permit in vivo purification of SHANK3-associated complexes. We will examine SHANK3 native complexes to determine whether there are distinct classes of SHANK3 protein complexes and the type of proteins in these sub-complexes. We will generate mice that either black or over-express Shank3 and we will characterize these Shank3 mouse models. In addition we will analyze behavioral, anatomical, and physiological consequences of modulating Shank3 expression in mice lacking FXR genes. Detailed behavioral and biochemical studies will allow us to establish a functional relationship between SHANK3 and FXR proteins in vivo and provide a foundation for detailed mechanistic studies that should benefit a broad population of ASD patients. PUBLIC HEALTH RELEVANCE: Autism is a heterogeneous disorder caused by mutations in different genes and is considered a major public health problem given the estimated prevalence of 1 in 150-200 children. We discovered that some autism- causing proteins interact with the same partners suggesting that there are shared pathways that lead to autism. In this grant, we will characterize some common pathways leading to autism, which would be of public health relevance because our studies will help many types of autism rather than one or two subtypes.

描述（由申请人提供）：该提案对RFA-MH-09-170，《恢复法》有限竞争的反应：解决自闭症谱系障碍异质性的研究（R01）。如RFA中指出的那样，自闭症谱系障碍（ASD）形成一个非常异构的群体：已确定30多个不同的基因是引起或促成自闭症原因。为了更好地理解这种多样化的基因如何引起类似的ASD表型，我们搜索了由自闭症相关基因编码的蛋白质的结合伙伴，并开发了自闭症蛋白质相互作用网络（一种“相互作用”），其中包含超过900蛋白质相互作用的蛋白质，其中许多是新颖的。这种相互作用组揭示的最令人兴奋的发现之一是发现有三种“集线器”蛋白是相互作用的中心：fxr1，shank3和tsc1。此外，shank3与两种不同类别的蛋白质相互作用：一种参与突触结构，另一种参与RNA代谢和翻译。后一类将shank3连接到脆弱的X蛋白（FMRP）及其旁系同源物（FXR1和2）以及结核性硬化症复合蛋白（TSC1和TSC2）。我们假设各种ASD之间存在共同的致病机制，其中一种以突触维持和可塑性为中心。我们建议Shank3在突触中扮演两个不同的角色，一个涉及突触组织和维护，另一个涉及突触时局部RNA翻译。为了深入了解ASD发病机理的关键机制，我们将识别并表征体内突触中的天然Shank3复合物。为此，我们将生成标记shank3蛋白的敲入小鼠，以允许体内纯化shank3相关的复合物。我们将检查shank3天然复合物，以确定这些亚复合物中是否存在不同类别的shank3蛋白复合物和蛋白质的类型。我们将生成黑色或过表达shank3的小鼠，我们将表征这些shank3鼠标模型。此外，我们将分析在缺乏FXR基因的小鼠中调节Shank3表达的行为，解剖和生理后果。详细的行为和生化研究将使我们能够在体内建立Shank3和FXR蛋白之间的功能关系，并为详细的机械研究提供基础，以使广泛的ASD患者受益。 公共卫生相关性：自闭症是由不同基因突变引起的一种异质性疾病，鉴于150-200名儿童的估计患病率估计为1个，被认为是一个主要的公共卫生问题。我们发现某些自闭症 - 导致蛋白质与同一伴侣相互作用，这表明有一些共同的途径导致自闭症。在这笔赠款中，我们将表征一些导致自闭症的常见途径，这将具有公共卫生相关性，因为我们的研究将帮助多种类型的自闭症，而不是一种或两个亚型。