Shank3 and the Par polarity complex in neurodevelopmental disorders

Shank3 和 Par 极性复合体在神经发育障碍中的作用

基本信息

批准号：
10390824
负责人：
Huaye Zhang
金额：
$ 41.3万
依托单位：
RBHS-ROBERT WOOD JOHNSON MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-15 至 2024-03-14
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10390824
关键词：
AMPA Receptors Actins Adaptor Signaling Protein Affect Behavioral Biochemical Biological Assay Biosensor Biotinylation Brain Cognitive Cognitive deficits Complement Complex Data Defect Dendritic Spines Disease Epithelial Cells Fluorescence Resonance Energy Transfer Genes Genetic Glutamates Hippocampus (Brain)Image Imaging Techniques Knock-in Mouse Light Link Mutant Strains Mice Mutate Mutation Neurodevelopmental Disorder Neurons PARD6A gene PHluorin Phosphorylation Proteins Regulation Resolution Role Scaffolding Protein Schizophrenia Signal Transduction Site Stimulus Structure Surface Symptoms Synapses Synaptic plasticity Testing Vertebral column autism spectrum disorder behavioral outcome behavioral phenotyping cognitive performance conditional knockout imaging approach in vivo live cell imaging molecular imaging mouse model mutant nanocluster recruit scaffold spatiotemporal synaptic function trafficking two-photon

项目摘要

Project Summary Autism Spectrum Disorders (ASD) and schizophrenia are two distinct neurodevelopmental disorders with certain shared symptoms. Yet the underlying synaptic mechanisms leading to these distinct and shared behavioral phenotypes remain elusive. Both disorders show a strong genetic component, and recent studies have pointed to a number of shared genes that are mutated in both ASD and schizophrenia. One of the most prominent monogenic genes for both disorders is SHANK3, which encodes a synaptic scaffolding protein. However, the mechanisms by which Shank3 mutations differentially affect synaptic functions are still unclear. A recent study in epithelial cells shows Shank2 interacts with the partitioning-defective (Par) polarity complex, which includes Par3, Par6 and atypical PKC (aPKC). Consistent with this, our preliminary data show that Shank3 interacts with aPKC in vivo, and this interaction is reduced in the Par3 conditional knockout hippocampus. Interestingly, we found that aPKC activation is reduced in Shank3 ASD-associated InsG3680 mutant mouse brains, while an overactivation of aPKC is observed in Shank3 schizophrenia-associated R1117X mutant brains. Thus, we hypothesize that Shank3 targets the Par polarity complex to synapses and regulates its activation. ASD- and schizophrenia-associated Shank3 mutations will result in differential dysregulation of aPKC activation, leading to distinct changes in synaptic plasticity and behavioral defects. In the first aim, we will test the hypothesis that dendritic spine structural plasticity is differentially affected by Shank3 ASD and schizophrenia mutants through dysregulated activation of aPKC. In Aim 2, we will test the hypothesis that dysregulation of aPKC activation in Shank3 mutants leads to altered AMPA receptor (AMPAR) trafficking and nanodomain localization at the spine surface. Our studies will utilize advanced molecular imaging techniques, including τSTED super resolution imaging, FRET and FRAP live cell imaging, combined with two-photon glutamate uncaging-induced dendritic spine structural plasticity. We will also complement these imaging approaches with biochemical analyses in SHANK3 ASD and schizophrenia- associated mutant knockin mouse models. Together, our proposed studies will establish a mechanistic link between Shank3 and the Par polarity complex in regulating synaptic plasticity in neurodevelopmental disorders including ASD and schizophrenia. As Par3 is genetically linked to ASD, schizophrenia, and high cognitive performance, our studies will shed light on the role of a Shank3-Par polarity signaling complex in cognitive defects in neurodevelopmental disorders.

项目概要自闭症谱系障碍 (ASD) 和精神分裂症是两种不同的神经发育障碍然而，导致这些独特和共同症状的潜在突触机制。这两种疾病的行为表型仍然难以捉摸，而且最近的研究表明，这两种疾病都有很强的遗传因素。指出了自闭症谱系障碍和精神分裂症中发生突变最多的一些共享基因。这两种疾病的主要单基因基因是 SHANK3，它编码突触支架蛋白。然而，Shank3 突变差异影响突触功能的机制仍不清楚。最近对上皮细胞的研究表明 Shank2 与分区缺陷 (Par) 极性复合物相互作用，其中包括 Par3、Par6 和非典型 PKC（aPKC），我们的初步数据表明： Shank3 在体内与 aPKC 相互作用，并且这种相互作用在 Par3 条件敲除中减少提示，我们发现 Shank3 ASD 相关的 InsG3680 中 aPKC 激活减少。突变小鼠大脑，而在与精神分裂症相关的 Shank3 中观察到 aPKC 过度激活 R1117X 突变大脑因此，我们追踪 Shank3 将 Par 极性复合体靶向突触。并调节其激活。 ASD 和精神分裂症相关的 Shank3 突变将导致 aPKC 激活的差异性失调，导致突触可塑性和突触可塑性的明显变化在第一个目标中，我们将检验树突棘结构可塑性的假设。 Shank3 ASD 和精神分裂症突变体通过 aPKC In 激活失调而受到不同程度的影响。目标 2，我们将测试 Shank3 突变体中 aPKC 激活失调导致改变的假设我们的研究将利用 AMPA 受体 (AMPAR) 运输和脊柱表面的纳米结构域定位。先进的分子成像技术，包括τSTED超分辨率成像、FRET和FRAP活细胞成像，结合双光子谷氨酸释放诱导的树突棘结构可塑性。还通过 SHANK3 ASD 和精神分裂症的生化分析来补充这些成像方法我们提出的研究将共同建立相关的突变敲入小鼠模型。 Shank3 和 Par 极性复合体在调节神经发育障碍中突触可塑性方面的作用包括自闭症谱系障碍 (ASD) 和精神分裂症。As Par3 与自闭症谱系障碍 (ASD)、精神分裂症和高认知能力有遗传相关。性能，我们的研究将揭示 Shank3-Par 极性信号复合物在认知中的作用神经发育障碍的缺陷。