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.

项目总结 迫切需要更深入地了解自闭症谱系的基本生物学机制。 疾病(ASD)是发现新的治疗靶点的途径。在这项提案中，我们的目标是利用削减- 边缘基因组技术结合斑马鱼的独特优势研究斑马鱼的损失效应 两个主要的ASD风险基因CHD8和SCN2A在发育中的大脑中的翻译调节功能。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，易发生细胞类型特异性缺陷。我们的中心假设是自闭症 基因破坏将导致发育中的脊椎动物大脑的整体翻译组发生变化，从而 到特定细胞类型的异常。这一假设是基于我们令人信服的初步数据 在三个顶端的斑马鱼突变体的全脑RNA-SEQ中，该翻译是一个显著失调的途径 ASD风险基因，所有这些基因都表现出健壮的行为、大脑结构和活动表型。为了测试这一点 假设，我们将使用一种新型的翻译核糖体亲和纯化测序(TRAP-SEQ) 斑马鱼Ribo-Tag转基因品系对全脑来源的神经元和胶质细胞中主动翻译的mRNAs的评价 斑马鱼CHD8和scn1lab突变体(目标1)和单细胞RNA-seq(scRNA-seq)的大脑特征 突变体中特定细胞类型的差异(目标2)。我们期待我们高度创新和探索性的方法 将为两个ASD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增进我们对 翻译作为自闭症的核心调控机制。这项研究将推动该领域的发展 阐明ASD基因下游具有潜在治疗意义的新的靶向途径。