Age-Dependent Dysfunction of GABAergic Neurotransmission Due to Autism-Associated mTOR Pathway Activation

自闭症相关 mTOR 通路激活导致 GABA 能神经传递的年龄依赖性功能障碍

• 批准号: 10090638
• 负责人: LAURA A JANSEN
• 金额: $ 39.38万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-18 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10090638
• 关键词: Adolescent; Adult; Age; Appearance; Brain; Cations; Child; Chlorides; Data; Dependence; Development; Disease; Down-Regulation; Electroencephalography; Environmental Risk Factor; Epilepsy; FRAP1 gene; Female; Functional disorder; Genetic; Human; Image; Immunohistochemistry; Impaired cognition; Impairment; Individual; Infant; Intervention; Lentivirus; Link; Maintenance; Mediating; Morbidity - disease rate; Mus; Mutation; Neonatal; Neuronal Plasticity; Neurons; PTEN gene; Pathologic; Pathway interactions; RE1-silencing transcription factor; Receptor Activation; Regulation; Rett Syndrome; Role; Seizures; Signal Transduction; Slice; TSC2 gene; Testing; Time; Transcription Repressor; Transgenic Mice; Up-Regulation; Western Blotting; age related; anxiety symptoms; associated symptom; autism spectrum disorder; comorbidity; course development; disability; gamma-Aminobutyric Acid; in vivo; mTOR Signaling Pathway; male; member; migration; neurotransmission; novel; patch clamp; premature; prevent; protein expression; receptor function; response; sex; synaptogenesis; voltage

Project Summary/Abstract Autism spectrum disorder (ASD) is a significant cause of morbidity and loss of developmental potential in children. An increasing number of genetic and environmental factors are being identified that contribute to ASD. Two major cellular pathways that have been implicated in both genetic and environmental causes of ASD involve mTOR signaling and GABAergic neurotransmission. In this project, we propose to investigate a mechanistic link between these pathways that may explain both the age dependence and male sex predominance of core ASD symptoms and comorbidities. In this project, we propose to test the hypothesis that autism-associated mTOR pathway activation causes differential dysregulation of GABAergic neurotransmission in the immature versus the mature brain. Further, we hypothesize that this dysregulation will be more pronounced in males than in females. We will examine our hypotheses through pursuit of the following Specific Aims. Specific Aim 1: To test the hypothesis that mTOR pathway activation accelerates the development of hyperpolarizing GABAA receptor function in the immature brain. Cultured cortical neurons and transgenic mice with mTOR pathway activation, either via heterozygous loss of the Pten or Tsc2 genes or via expression of a constitutively active form of mTOR, will be used. Specific Aim 2: To test the hypothesis that mTOR pathway activation interferes with maintenance of hyperpolarizing GABAA receptor function in the mature brain. In Specific Aim 2, the studies of Aim 1 will be repeated in mature neuronal cultures and in adult mice. Specific Aim 3: To define the role of the transcriptional repressor REST (RE1-Silencing Transcription factor) in the age-dependent dysregulation of GABAergic neurotransmission by mTOR pathway activation. We will utilize cortical neuronal cultures and lentivirus-mediated manipulation of REST function in vivo to investigate the involvement of REST in the bidirectional regulation of KCC2 expression by mTOR pathway activation. As a result of these studies, we will be able to identify the differential effects of mTOR pathway activation on inhibitory neurotransmission in immature versus mature and in male versus female brain, thereby suggesting pathways for development of novel, targeted interventions for ASD and its comorbidities.

项目总结/摘要 自闭症谱系障碍（ASD）是一个重要的原因，发病率和损失的发展， 孩子的潜力。越来越多的遗传和环境因素正在被确定， 为ASD做贡献两个主要的细胞途径，涉及遗传和 ASD的环境原因涉及mTOR信号传导和GABA能神经传递。在本项目中， 我们建议调查这些途径之间的机械联系，这可能解释年龄和年龄之间的关系。 核心ASD症状和合并症的依赖性和男性优势。 在这个项目中，我们打算检验自闭症相关mTOR通路激活的假设， 导致未成熟与成熟的GABA能神经传递的差异失调 个脑袋此外，我们假设这种失调在男性中比在女性中更明显。 我们将通过追求以下具体目标来检验我们的假设。具体目标1：测试 mTOR通路激活加速超极化GABAA发展的假设 未成熟大脑中的受体功能培养的皮层神经元和具有mTOR的转基因小鼠 通过Pten或Tsc 2基因的杂合丢失或通过Pten或Tsc 2基因的表达， 将使用mTOR的组成型活性形式。具体目标2：检验mTOR 通路激活干扰成熟神经元中超极化GABAA受体功能的维持 个脑袋在特定目标2中，将在成熟神经元培养物和成人中重复目标1的研究。 小鼠具体目标3：确定转录抑制因子REST（RE 1-沉默）的作用 转录因子）在mTOR对GABA能神经传递的年龄依赖性失调中的作用 通路激活我们将利用皮层神经元培养和慢病毒介导的操纵， REST在体内的功能，以研究REST参与KCC 2的双向调节 通过mTOR通路激活表达。 作为这些研究的结果，我们将能够确定mTOR通路的差异效应， 在未成熟与成熟以及男性与女性大脑中对抑制性神经传递的激活， 从而为ASD及其并发症的新型靶向干预的开发提供了途径。 合并症。