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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要自闭症谱系障碍 (ASD) 是发病和发育障碍的一个重要原因儿童的潜力。越来越多的遗传和环境因素被发现为自闭症谱系障碍做出贡献。与遗传和遗传有关的两条主要细胞途径 ASD 的环境原因涉及 mTOR 信号传导和 GABA 神经传递。在这个项目中，我们建议研究这些途径之间的机制联系，这可能解释年龄自闭症谱系障碍 (ASD) 核心症状和合并症的依赖性和男性占主导地位。在这个项目中，我们建议检验以下假设：自闭症相关的 mTOR 通路激活导致未成熟与成熟的 GABA 能神经传递的差异失调脑。此外，我们假设这种失调在男性中比在女性中更为明显。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。培养皮层神经元和带有 mTOR 的转基因小鼠途径激活，或者通过 Pten 或 Tsc2 基因的杂合缺失，或者通过表达将使用 mTOR 的组成型活性形式。具体目标 2：检验 mTOR 的假设通路激活干扰成熟细胞中超极化 GABAA 受体功能的维持脑。在具体目标 2 中，目标 1 的研究将在成熟神经元培养物和成人中重复进行。老鼠。具体目标 3：确定转录阻遏蛋白 REST（RE1-Silencing mTOR 对 GABA 能神经传递的年龄依赖性失调中的转录因子）通路激活。我们将利用皮质神经元培养和慢病毒介导的操作 REST 体内功能研究 REST 参与 KCC2 双向调节通过 mTOR 通路激活表达。通过这些研究，我们将能够确定 mTOR 通路的差异效应未成熟与成熟以及男性与女性大脑中抑制性神经传递的激活，从而为自闭症谱系障碍（ASD）及其相关疾病的新型、有针对性的干预措施的开发提出了途径合并症。