Elucidating the synaptic interactome of the high risk autism gene ANK2

阐明高风险自闭症基因 ANK2 的突触相互作用组

基本信息

批准号：
10391766
负责人：
Damaris N Lorenzo
金额：
$ 45.17万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-07 至 2022-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10391766
关键词：
ANK2 gene ASD patient Action Potentials Affect Anatomy Antibodies Architecture Axon Binding Biological Assay Biology Biotin Brain Cells Cellular Assay Complex Couples Cytoskeletal Proteins Dendrites Dendritic Spines Development Exhibits Functional disorder Genes Glutamates Goals Image Impairment In Vitro Ion Channel Knock-in Mouse Knockout Mice Label Ligase Link Mass Spectrum Analysis Membrane Membrane Proteins Membrane Transport Proteins Microscopy Molecular Morphology Motor Mus Neurons Pathway Analysis Pathway interactions Phenocopy Play Positioning Attribute Protein Isoforms Proteins Proteomics Resolution Role Sodium Channel Synapses Variant Vertebral column Vesicle autism spectrum disorder base chromatin remodeling density disorder risk excitatory neuron experimental study falls gene network high risk hippocampal pyramidal neuron in vivo individuals with autism spectrum disorder nanoscale novel postsynaptic postsynaptic density protein protein complex scaffold synaptic function tandem mass spectrometry trafficking voltage

项目摘要

PROJECT SUMMARY Network gene analysis suggests that high confidence autism spectrum disorder (hcASD) genes operate through convergent mechanisms that predominantly support synaptic function, which, in turn, can determine anatomical and functional brain connectivity. Several variants in ANK2 have been identified in ASD patients and this gene has been consistently considered a top hcASD risk. ANK2 encodes ankyrin-B (AnkB), which roles in axonal biology may be an underlying factor in ASD pathophysiology. However, our recent findings indicate that ankyrin- B also plays important roles at the cortical excitatory postsynapse. Moreover, both predominant AnkB isoforms in neurons (220 kDa and 440 kDa AnkB) are abundantly expressed in dendrites, where they likely perform isoform-specific roles and interact with unique partners, including proteins encoded by other hsASD genes. Thus, it is important to define the isoform-specific roles and partners of AnkB at the postsynapse. To accomplish these goal we will 1) use in vivo proximity-dependent biotin identification (iBioID) combined with in tandem mass spectrometry (MS) to capture and identify AnkB interacting partners in the postsynapse by leveraging our novel biotin ligase BirA-tagged conditional AnkB (AnkB-BioID) knock-in mouse and isoform-specific AnkB knockout mice; 2) combine live and super-resolution microscopies and cellular assays to validate compelling AnkB interactors; and 3) define the significance of novel AnkB PSD complexes for synaptic development and function.

项目摘要网络基因分析表明，高置信自闭症谱系障碍（HCASD）基因通过主要支持突触功能的收敛机制，这又可以确定解剖学和功能性大脑连通性。在ASD患者和该基因中已经确定了ANK2中的几种变体一直被认为是最高的HCASD风险。 ANK2编码Ankyrin-B（ANKB），在轴突中扮演角色生物学可能是ASD病理生理学的根本因素。但是，我们最近的发现表明脚踝蛋白 B还在皮质兴奋性后的结合后发挥了重要作用。此外，这两个主要ANKB同工型在神经元（220 kDa和440 kDa ankb）中，在树突中大量表达，它们可能会执行同工型特异性角色并与独特的伴侣相互作用，包括由其他HSASD基因编码的蛋白质。因此，在节奏后，定义ANKB的同工型特异性角色和伴侣很重要。完成这些目标我们将1）在体内接近依赖性生物素鉴定（IBIOID）中使用串联质量光谱法（MS）通过利用我们的小说来捕获和识别后共同伴侣的ANKB相互作用伙伴生物素连接酶BIRA标记的条件ANKB（ANKB生命）敲入鼠标和同工型特异性ANKB敲除小鼠； 2）结合现场和超分辨率显微镜和细胞测定，以验证引人注目的ANKB 互动者； 3）定义了新型ANKB PSD复合物对突触发育和功能的重要性。