Cellular and molecular mechanisms disrupted in 22q13 deletion syndrome and autism

22q13 缺失综合征和自闭症的细胞和分子机制被破坏

• 批准号: 10084752
• 负责人: Oleksandr Shcheglovitov
• 金额: $ 38.13万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10084752
• 关键词: AMPA Receptors; Address; Affect; Alleles; Astrocytes; Back; Basic Science; Behavioral; Binding; Biochemistry; Brain; Cell Line; Chronic; Complex; Cytoskeleton; Data; Defect; Development; Electrophysiology (science); Endocytosis; Engraftment; Etiology; Excitatory Synapse; Financial compensation; Functional disorder; Gene Expression; Genes; Genetic; Genetic Engineering; Genetic Suppression; Goals; HCN1 gene; Human; Image; Impairment; In Vitro; Induced pluripotent stem cell derived neurons; Intellectual functioning disability; Knock-out; Laboratories; Lead; Measures; Mediating; Membrane; Mental disorders; Mission; Modeling; Molecular; Molecular Abnormality; Mus; Mutate; Mutation; National Institute of Mental Health; Neurodevelopmental Disorder; Neurons; Pathology; Patients; Pharmacology; Phelan-McDermid syndrome; Phenotype; Prefrontal Cortex; Property; Proteins; Public Health; Reporting; Research; Research Project Grants; Resistance; Rodent; Role; Scaffolding Protein; Synapses; Synaptic Transmission; Syndrome; Techniques; Technology; Testing; Translations; Up-Regulation; autism spectrum disorder; brain abnormalities; engineered stem cells; experimental study; functional disability; improved; in vivo; individuals with autism spectrum disorder; induced pluripotent stem cell; insight; knock-down; neural network; novel; novel therapeutic intervention; novel therapeutics; therapeutically effective

Mutations in genes encoding synaptic proteins and impaired functional brain connectivity are emerging as common deficits associated with autism spectrum disorders (ASDs). However, how mutated synaptic proteins affect the properties of human neurons at the cellular and molecular levels to cause abnormal brain connections remains an important unanswered question. Addressing this question is essential for understanding the etiology and pathology of ASDs and developing novel and effective therapeutic strategies for patients. The PI previously demonstrated that SHANK3-deficient human cortical neurons derived from induced pluripotent stem cells (iPSCs), generated from 22q13 deletion syndrome patients with autism, have severely impaired intrinsic excitability and excitatory synaptic transmission. However, how these two phenotypes develop and affect neuronal connectivity in the brain remain unknown. The main goal of this project is to elucidate the cellular and molecular mechanisms responsible for development of synaptic and connectivity deficits in SHANK3-deficient human neurons. SHANK3 is a scaffolding protein expressed at excitatory synapses that have been frequently found to be mutated or deleted in individuals with autism and intellectual disability. The central hypothesis of this project is that synaptic deficits in SHANK3-deficient human neurons develop as a result of elevated electrical activity and activity-mediated weakening and elimination of excitatory synapses. This hypothesis is strongly supported by the preliminary data obtained in the PI's laboratory. The following Specific Aims are formulated to test this hypothesis: 1) Determine the role of elevated electrical activity in development of synaptic deficits in SHANK3-deficient human neurons; 2) Determine the role of ARC in development of synaptic deficits in SHANK3-deficient human neurons; and 3) Determine how loss of SHANK3 in human neurons impacts synaptic inputs onto these neurons in vivo. Under these aims, the properties of human cortical neurons generated from novel, precisely genetically-engineered stem cell lines will be investigated using electrophysiology, imaging, and biochemistry techniques in vitro and upon engraftment into the mouse brain. The proposed research is significant because it is expected to substantially advance understanding of the molecular, cellular, and circuitry mechanisms disrupted in autism and intellectual disability, and guide the development of novel therapeutic strategies for patients.

编码突触蛋白的基因突变和大脑功能连接受损正在出现 作为与自闭症谱系障碍 (ASD) 相关的常见缺陷。然而，突触如何突变 蛋白质在细胞和分子水平上影响人类神经元的特性，导致大脑异常 连接仍然是一个重要的悬而未决的问题。解决这个问题至关重要 了解自闭症谱系障碍的病因和病理学并制定新颖有效的治疗策略 对于患者。 PI 之前证明，SHANK3 缺陷的人类皮质神经元源自 诱导多能干细胞（iPSC）由 22q13 缺失综合征自闭症患者产生， 内在兴奋性和兴奋性突触传递严重受损。然而，这两个如何 表型的发展和影响大脑神经元连接的情况仍然未知。此次活动的主要目标 该项目旨在阐明负责突触和突触发育的细胞和分子机制 SHANK3 缺陷的人类神经元的连接缺陷。 SHANK3 是一种支架蛋白，表达于 经常发现自闭症患者的兴奋性突触发生突变或删除 智力障碍。该项目的中心假设是 SHANK3 缺陷人类的突触缺陷 神经元的发育是电活动升高和活动介导的弱化和消除的结果 兴奋性突触。这一假设得到 PI 中获得的初步数据的有力支持 实验室。制定以下具体目标来检验这一假设： 1) 确定升高的作用 SHANK3 缺陷的人类神经元突触缺陷发展中的电活动； 2）确定 ARC 在 SHANK3 缺陷的人类神经元突触缺陷发展中的作用； 3) 确定如何 人类神经元中 SHANK3 的缺失会影响体内这些神经元的突触输入。在这些目标下， 由新颖的、精确的基因工程干细胞系产生的人类皮质神经元的特性将 在体外和植入后使用电生理学、成像和生物化学技术进行研究 进入小鼠大脑。拟议的研究意义重大，因为预计它将大大推进 对自闭症和智力障碍的分子、细胞和电路机制的理解 残疾，并指导为患者制定新的治疗策略。