Neuronal and Behavioral Deficits Associated with Scn2a Deficiency in Autism Spectrum Disorder

自闭症谱系障碍中与 Scn2a 缺乏相关的神经元和行为缺陷

• 批准号: 10211758
• 负责人: Yang Yang
• 金额: $ 38.15万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10211758
• 关键词: Address; Adult; Affect; Agonist; Animal Behavior; Behavior; Behavioral; Behavioral Genetics; Brain; Characteristics; Child; Data; Development; Disease; Electrodes; Electrophysiology (science); Elements; Equilibrium; Frequencies; Genes; Genetic; Genetic study; Human; Impairment; Interneurons; Intervention; Knock-out; Knockout Mice; Knowledge; Length; Medial; Mission; Modeling; Mus; Mutate; Mutation; Neurodevelopmental Disorder; Neurons; Outcome; Pathway interactions; Perinatal; Pharmacology; Predisposition; Prefrontal Cortex; Property; Proteins; Research Personnel; Resolution; Rest; Signal Transduction; Slice; Social Behavior; Social Interaction; Sodium Channel; Synapses; TNFSF5 gene; Technology; Testing; United States; United States National Institutes of Health; Work; anxiety-like behavior; autism spectrum disorder; awake; behavioral impairment; behavioral outcome; behavioral response; behavioral study; clinically relevant; density; efficacy evaluation; excitatory neuron; gamma-Aminobutyric Acid; hippocampal pyramidal neuron; in vivo; innovation; insight; interest; loss of function mutation; mouse model; novel; novel therapeutic intervention; optogenetics; patch clamp; postsynaptic; public health relevance; receptor; response; restoration; social; social communication impairment; social deficits; spatiotemporal; synaptic function; therapeutic development; therapy development; transcriptome sequencing; voltage

Project Summary/Abstract Autism spectrum disorder (ASD) is a neurodevelopmental disorder associated with impaired social communications and behavioral abnormalities, which affects ~1 in 54 children in the United States (CDC.gov). SCN2A, encoding neuronal voltage-gated sodium channel Nav1.2, has been identified as one of the leading genes associated with ASD. We have characterized a novel Scn2a-deficient mouse model that is generated via targeted gene-trap knockout (gtKO) strategy and possesses a built-in genetic rescue element. Our preliminary data revealed profound behavioral abnormalities in homozygous Scn2agtKO/gtKO mice including anxiety-like behaviors, impaired nesting and social deficits. We also identified elevated excitation-inhibition (E/I) balance in pyramidal neurons of mPFC, which has been implicated in ASD and social deficits. However, a critical gap exists regarding how in vivo neuronal firings in mPFC are affected by elevated E/I balance and to what extent the manipulation of E/I balance will alter the behavioral outcomes in Scn2agtKO/gtKO mice. To address this gap, we propose to test an overarching hypothesis that Scn2a deficiency increases E/I balance, impairs neuronal responses in mPFC, and results in social deficits that can be rescued with targeted genetic and pharmacological interventions. In Aim 1, we will assess the synaptic properties and in vivo firing of neurons in mPFC using brain- slice patch-clamp recording and Neuropixels in vivo recording. Our findings are expected to provide cellular-level and local network level neuropathological mechanisms of Scn2a deficiency. In Aim 2, we will determine neuronal firings and behavioral outcomes in response to manipulating E/I balance in mPFC microcircuit using optogenetics and chemogenetics. Our findings will bolster the significance of E/I balance modulation for correction of behavioral deficits. In Aim 3, we will evaluate the efficacy of timed genetic and pharmacological rescue to determine optimal windows for intervention. Our study is significant in the following ways: i) SCN2A deficiency to be studied is among the leading monogenetic forms of ASD; ii) Excitation and inhibition (E/I) balance of mPFC microcircuit to be thoroughly dissected is closely associated with social deficits; and iii) Genetic rescue and pharmacological intervention to be tested are of clear clinical relevance, and will provide translational basis to inform therapeutic development for the treatment of Scn2a-deficiency related disorders. Our study has the following innovations: i) use of novel Scn2agtKO/gtKO mice that display profound cellular and behavioral deficits; ii) innovative ways to achieve genetic and pharmacological rescue; and iii) use of cutting-edge technologies including high density Neuropixels in vivo recordings. The applicant is an early stage investigator (ESI), whose team has extensive expertise in sodium channel electrophysiology, animal behaviors, genetics and pharmacology. The team is well suited to carry out the proposed work to its full completion within the project timeframe, and generate impactful outcomes to advance the field.

项目摘要/摘要 自闭症谱系障碍（ASD）是一种与社会受损相关的神经发育障碍 沟通和行为异常，在美国54名儿童中有〜1个（CDC.GOV）。 编码神经元电压门控钠通道NAV1.2的SCN2A已被确定为领先的 与ASD相关的基因。我们已经表征了一种新型的SCN2A缺陷小鼠模型，该模型是通过 靶向基因陷阱敲除（GTKO）策略，并具有内置的遗传救援元件。我们的初步 数据揭示了纯合SCN2AGTKO/GTKO小鼠的深刻行为异常，包括焦虑 行为，筑巢和社会缺陷受损。我们还确定了激发抑制（E/I）的升高 MPFC的锥体神经元，与ASD和社会缺陷有关。但是，存在关键的差距 关于MPFC中的体内神经元射击如何受到E/I平衡升高以及在多大程度的影响 E/I平衡的操纵将改变SCN2AGTKO/GTKO小鼠中的行为结果。为了解决这个差距，我们 提议检验总体假设，即SCN2A缺乏会增加E/I平衡，损害神经元 MPFC的反应，并导致可以通过有针对性的遗传和药理学来挽救的社会缺陷 干预措施。在AIM 1中，我们将使用脑部评估MPFC中神经元的突触性能和体内发射 切片片钳记录和体内记录中的神经质子。我们的发现有望提供蜂窝级 SCN2A缺乏的局部网络水平神经病理机制。在AIM 2中，我们将确定神经元 使用光遗传学在MPFC微电路中操纵E/I平衡的解雇和行为结果 和化学遗传学。我们的发现将加强E/I平衡调制的意义 行为不足。在AIM 3中，我们将评估定时遗传和药理救援的功效 确定干预的最佳窗口。我们的研究以以下方式很重要：i）SCN2A缺乏症 被研究是ASD的主要单基因形式之一。 ii）MPFC的激发和抑制（E/I）平衡 要彻底剖析的微电路与社会缺陷密切相关。和iii）遗传救援和 要测试的药理干预措施具有明显的临床相关性，并将为 为治疗与SCN2A缺乏障碍的治疗提供信息。我们的研究有 经过创新之后：i）使用新型的SCN2Agtko/GTKO小鼠，这些小鼠表现出深远的细胞和行为缺陷； ii） 实现遗传和药理学救援的创新方法； iii）使用尖端技术 包括体内记录中的高密度神经质子。申请人是早期的调查员（ESI），他的 团队在钠通道电生理学，动物行为，遗传学和 药理。该团队非常适合在项目中进行全面完成拟议的工作 时间范围，并产生有影响力的成果以推进该领域。