TSC and sensory processing

TSC 和感官处理

• 批准号: 9892431
• 负责人: Angelique Bordey
• 金额: $ 25.13万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-15 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9892431
• 关键词: Address; Adult; Affect; Autophagocytosis; Axon; Biological Models; Brain; CRISPR/Cas technology; Cells; Child; Cognitive deficits; Cortical Dysplasia; Data; Defect; Dendrites; Dependence; Development; Disease; Electroporation; Embryo; FRAP1 gene; Goals; Human; Hyperactive behavior; Individual; Lateral; Lead; Length; Life; Link; Mendelian disorder; Molecular; Molecular Abnormality; Morphology; Mus; Mutation; Neurodevelopmental Disorder; Neurologic Deficit; Neurons; Pathway interactions; Pattern; Publishing; Reporting; Seizures; Sensory; Signal Transduction; Sirolimus; Somatic Mutation; Somatosensory Cortex; Synapses; TSC1 gene; TSC2 gene; Testing; Thalamic structure; Time; Translations; Tuberous sclerosis protein complex; Vibrissae; Work; autism spectrum disorder; barrel cortex; base; brain abnormalities; in utero; information processing; mTOR Signaling Pathway; mTOR inhibition; malformation; migration; mutant; postnatal; prevent; response; tool; vector

The goal of this proposal is to identify how abnormalities of brain development in monogenic disorders involving the PI3K-mTOR pathway (e.g., tuberous sclerosis complex [TSC]) lead to alterations in circuit and in information processing. TSC is a classical disorder of the mTOR signaling pathway due to second-hit somatic mutations in TSC1 or TSC2 leading to mTOR hyperactivity and developmental malformations associated with seizures and neurological deficits. 50-60% of children with TSC have severe cognitive deficits and autism accompanied with abnormal sensory processing. Published work from our lab identified a combination of molecular and cellular alterations in murine neurons with hyperactive mTOR. These alterations include neuron misplacement and dysmorphogenesis that are conserved across cortical regions and between mice and humans. However, the impact of these defects on circuit formation and information processing, and the dependence on mTOR and downstream pathways at the circuit level remain unclear. Here, we propose to use the mouse barrel cortex as a well-established model system to start addressing the link between neuronal abnormalities and alterations in circuitry and sensory processing. Building from our previous findings, we hypothesize that dysmorphogenesis of layer (L) 4 barrel cortex neurons in TSC contributes to mTOR-dependent abnormalities in circuitry, functional connectivity, and sensory responses. We have the following two aims: (1) To test the hypothesis that mTOR hyperactivity in L4 neurons alters barrel circuitry and functional connectivity. (2) To test the hypothesis that reducing mTOR activity during a brief critical window and normalizing two downstream pathways prevent abnormalities in circuit and sensory responses in TSC condition. To address these aims, we will use in utero electroporation to express RhebCA and generate humanized Tsc1 mutations using CRISPR/Cas9 in L4 neurons of the barrel cortex.

这项建议的目的是确定如何在单基因的大脑发育异常， 涉及PI 3 K-mTOR通路的疾病（例如，结节性硬化症（TSC）导致 电路和信息处理的改变。TSC是mTOR的经典病症， 由于TSC 1或TSC 2中的二次击中体细胞突变导致mTOR的信号传导途径 与癫痫和神经系统疾病相关的多动症和发育畸形 赤字50-60%的TSC儿童有严重的认知缺陷和自闭症， 感觉处理异常我们实验室发表的研究发现， 以及mTOR过度活跃的鼠神经元中的细胞改变。这些变化包括 神经元错位和形态发育异常，这些在皮层区域中是保守的， 老鼠和人类之间的关系。然而，这些缺陷对电路形成和 信息处理，以及对mTOR和回路下游通路的依赖性 水平仍不清楚。在这里，我们建议使用小鼠桶皮质作为一个公认的 模型系统开始解决神经元异常和改变之间的联系， 电路和感觉处理。根据我们之前的发现，我们假设， TSC中第4层（L）桶皮质神经元的畸形发生有助于mTOR依赖性的 电路、功能连接和感觉反应的异常。我们有以下 两个目的：（1）验证L4神经元中mTOR过度活跃改变桶回路的假设 和功能连接。(2)为了验证在短暂的睡眠中降低mTOR活性 临界窗口和正常化两个下游通路防止电路异常， TSC条件下的感觉反应。为了实现这些目标，我们将使用子宫内电穿孔 在L4神经元中表达RhebCA并使用CRISPR/Cas9产生人源化Tsc 1突变 大脑皮层