Investigating the Translatome in Genetic Models of Autism

研究自闭症遗传模型中的翻译组

• 批准号: 10649109
• 负责人: ELLEN J HOFFMAN
• 金额: $ 25.02万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-15 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10649109
• 关键词: Affect; Affinity Chromatography; Behavioral; Biological; Brain; Cell physiology; Communication; Data; Development; Disparate; FMR1; FMRP; Functional disorder; Gene Expression Regulation; Gene set enrichment analysis; Genes; Genetic Models; Genomics; Goals; Knowledge; Lead; Mammals; Messenger RNA; Molecular; Neuroglia; Neurons; Orthologous Gene; Outcome; Outcomes Research; Pathway interactions; Phenotype; Process; Proliferating; Protein Biosynthesis; Research; RiboTag; Ribosomes; Risk; Role; Technology; Testing; Therapeutic; Transgenic Organisms; Translating; Translational Regulation; Translations; Zebrafish; autism spectrum disorder; cell type; differential expression; gene discovery; gene function; gliogenesis; in vivo; innovation; insight; loss of function; mutant; neural; neurogenesis; new therapeutic target; novel; risk variant; single-cell RNA sequencing; transcriptome sequencing; translatome

PROJECT SUMMARY There is an urgent need to gain a greater understanding of basic biological mechanisms in autism spectrum disorders (ASDs) as a path to discover new therapeutic targets. In this proposal, we aim to leverage cutting- edge genomic technologies together with the unique advantages of zebrafish to investigate the effect of loss of function of two top ASD risk genes, CHD8 and SCN2A, on translation regulation in the developing brain. Our central goal is to elucidate the mechanisms by which loss of function of these genes affects translation in both neurons and glia, predisposing to cell type-specific deficits. Our central hypothesis is that ASD gene disruption will lead to alterations in the global translatome in the developing vertebrate brain, predisposing to cell type-specific abnormalities. This hypothesis is based on our compelling preliminary data demonstrating that translation is a significantly dysregulated pathway in whole-brain RNA-seq of zebrafish mutants of three top ASD risk genes, all of which display robust behavioral, brain structural and activity phenotypes. To test this hypothesis, we will perform Translating Ribosome Affinity Purification sequencing (TRAP-seq) using a novel zebrafish Ribo-Tag transgenic line to evaluate actively translated mRNAs in neurons and glia derived from whole brains of zebrafish mutants of chd8 and scn1lab (Aim 1), and single cell RNA-seq (scRNA-seq) to characterize cell type-specific differences in mutants (Aim 2). We expect that our highly innovative and exploratory approach will provide new insights into how two top ASD risk genes affect translation regulation and cell type composition in the developing vertebrate brain. The expected outcome of this research is to advance our understanding of translation as a central regulatory mechanisms underlying ASDs. This research will advance the field by illuminating a novel targetable pathway downstream of ASD genes with potential therapeutic implications.

项目概要 迫切需要更好地了解自闭症谱系的基本生物学机制 障碍（ASD）作为发现新治疗靶点的途径。在本提案中，我们的目标是利用削减 边缘基因组技术与斑马鱼的独特优势一起研究缺失的影响 两个顶级 ASD 风险基因 CHD8 和 SCN2A 在发育中大脑翻译调节中的功能。我们的 中心目标是阐明这些基因功能丧失影响翻译的机制 在神经元和神经胶质细胞中，容易出现细胞类型特异性缺陷。我们的中心假设是 ASD 基因破坏将导致发育中的脊椎动物大脑中的整体翻译组发生改变，从而诱发 细胞类型特异性异常。这个假设是基于我们令人信服的初步数据证明 在三个顶级斑马鱼突变体的全脑 RNA-seq 中，翻译是一个显着失调的途径 自闭症谱系障碍（ASD）风险基因，所有这些基因都表现出强大的行为、大脑结构和活动表型。为了测试这个 假设，我们将使用一种新颖的方法进行翻译核糖体亲和纯化测序（TRAP-seq） 斑马鱼 Ribo-Tag 转基因系，用于评估来自整个细胞的神经元和神经胶质细胞中活跃翻译的 mRNA chd8 和 scn1lab 斑马鱼突变体的大脑（目标 1）和单细胞 RNA-seq (scRNA-seq) 来表征 突变体中细胞类型特异性的差异（目标 2）。我们期望我们高度创新和探索性的方法 将为两个自闭症谱系障碍（ASD）顶级风险基因如何影响翻译调控和细胞类型组成提供新的见解 在发育中的脊椎动物大脑中。这项研究的预期结果是加深我们对 翻译为自闭症谱系障碍（ASD）背后的中央监管机制。这项研究将通过以下方式推进该领域的发展 阐明了 ASD 基因下游的一条新的靶向途径，具有潜在的治疗意义。