Molecular dissection of synaptic dysfunction in mental disorders

精神疾病突触功能障碍的分子解剖

基本信息

批准号：
10332733
负责人：
Changhui Pak
金额：
$ 45.24万
依托单位：
UNIVERSITY OF MASSACHUSETTS AMHERST
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-01 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10332733
关键词：
3-Dimensional Affect Animals Astrocytes Autopsy Biochemistry Biological Bipolar Disorder Brain Brain Diseases Calcium Calmodulin Cell Adhesion Molecules Cells Characteristics Cognition Disorders Congenital cerebellar hypoplasia Defect Development Disease Disease model Dissection Dopamine Electrophysiology (science)Engineering Event Experimental Designs Forebrain Development Functional disorder Gender Genetic Genome engineering Genomics Glutamates Goals Grant Human Image Immunoprecipitation Impairment In Vitro Intellectual functioning disability Invertebrates Knock-out Lead Link Maintenance Mass Spectrum Analysis Mediating Mental disorders Methodology Methods Microcephaly Modeling Molecular Molecular Profiling Mood Disorders Mus Mutation Nature Neuronal Differentiation Neurons Neurosciences Pathogenesis Pathology Pathway interactions Patients Perinatal mortality demographics Pharmaceutical Preparations Phenotype Pontine structure Probability Protein Kinase Protein Kinase Interaction Protein-Serine-Threonine Kinases Proteins Proxy Reporting Research Risk Factors Role Scaffolding Protein Schizophrenia Signal Transduction Societies Synapses Synaptic Transmission Techniques Testing Tissues Translating Up-Regulation Viral Work X-linked intellectual disability autism spectrum disorder cell type conditional knockout excitatory neuron genetic analysis genetic variant genome sequencing genome-wide human embryonic stem cell immunocytochemistry induced pluripotent stem cell knockout gene live cell imaging loss of function mutant nerve stem cell neurodevelopment neuropsychiatric disorder neurotransmitter release novel novel therapeutics overexpression presynaptic risk variant schizophrenia risk single-cell RNA sequencing stem stem cells synaptic function synaptogenesis tool

项目摘要

Genetic analyses of polygenic brain disorders, such as autism spectrum disorders (ASDs) and schizophrenia (SZ), have revealed “synaptic dysfunction” as a key cellular substrate for these disorders. Yet, translating this synaptic hypothesis to in vitro disease modeling and extracting disease-relevant biological information has been challenging. The PI has previously established a human neuronal model of SZ, combining isogenic genome engineering, patient-derived induced pluripotent stem cells (iPSCs), and induced neuronal differentiation (iN cells). Human neurons bearing mutations in the synaptic cell adhesion molecule Neurexin-1 (NRXN1, 2p16.3), a bona fide risk allele for SZ, display deficits in excitatory synaptic strength and neurotransmitter release probability as well as a consistent upregulation of calcium/calmodulin-dependent serine protein kinase (CASK) protein level, by which the mechanism is currently not understood. It remains unclear how these phenotypes arise and lead to synaptic pathology and abnormal neuronal networks implicated in the disease. In this grant, we aim to understand the cell type- and developmental-specific functions of NRXN1-CASK interaction in normal synapse development and their mechanistic contributions to SZ. Using synaptic molecules as a proxy, we will dissect how disruptions in the synaptic pathway can prime or actively participate in SZ. We will achieve this by studying aberrant CASK signaling in NRXN1 mutant background (Aim 1), investigating CASK’s normal function at human synapses using CASK KO induced neurons (Aim 2) and by revealing key cellular events mediated by NRXN1 and CASK during human forebrain development using KO cortical spheroid models (Aim 3). Our work integrates techniques in imaging, electrophysiology, biochemistry and single cell RNA-seq with rigorous experimental designs using isogenic engineered and patient-derived iPSCs from multiple genetic and gender backgrounds, differentiation of pure induced neuronal subtypes with defined synaptic characteristics and differentiation of 3-D cortical spheroids with characterized cellular features. Findings of this grant will provide mechanistic understanding of the molecular and cellular underpinnings of neuropsychiatric disorders and such information will translate to other disorders of the synapse, including intellectual disability, ASDs, and bipolar and mood disorders.

多基因脑疾病的遗传分析，例如自闭症谱系障碍（ASDS）和精神分裂症（SZ）已揭示了“突触功能障碍”作为这些疾病的关键细胞底物。但是，翻译这个突触假说是体外疾病建模和提取与疾病相关的生物学信息的假设他具有挑战性。 PI以前已经建立了SZ的人类神经元模型，结合了同源性基因组工程，患者衍生的诱导多能干细胞（IPSC），并诱导神经元分化（在细胞中）。突触细胞粘附分子神经1中有突变的人神经元（NRXN1，2P16.3），SZ的真正风险等位基因，显示在兴奋性突触强度和神经递质释放概率以及钙/钙调蛋白依赖性的一致上调序列蛋白激酶（桶）蛋白水平，目前尚不清楚该机制。它仍然存在尚不清楚这些表型如何出现并导致突触病理和异常神经元网络在疾病中实施。在这笔赠款中，我们旨在了解细胞类型和特定于发育的细胞类型 NRXN1掩盖相互作用在正常突触发育中的功能及其机械贡献 SZ。使用合成分子作为代理，我们将剖析突触途径中的破坏如何质量或积极参与SZ。我们将通过研究NRXN1突变体中的异常桶信号来实现这一目标背景（AIM 1），使用木桶ko诱导的木桶在人类突触中的正常功能神经元（AIM 2）和通过揭示由NRXN1和CASK介导的人类前脑介导的关键细胞事件使用KO皮质球体模型开发（AIM 3）。我们的工作将技术整合到成像中，具有严格的实验设计的电生理学，生物化学和单细胞RNA-seq使用同源性设计来自多种遗传和性别背景的工程和患者衍生的IPSC，纯粹的分化具有定义的合成特性和3-D皮层的分化的诱导神经元亚型具有特征性的细胞特征。这项赠款的发现将提供对神经精神疾病的分子和细胞基础，这些信息将转化为其他突触的疾病，包括智力残疾，ASD，双相情绪和情绪障碍。