Systematic functional study of 21st chromosome ortholog overexpression in C. elegans

线虫第 21 号染色体直系同源物过表达的系统功能研究

基本信息

批准号：
10651500
负责人：
JONATHAN THOMAS PIERCE
金额：
$ 6.62万
依托单位：
UNIVERSITY OF TEXAS AT AUSTIN
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-06-01 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10651500
关键词：
Address Alzheimer&apos s Disease Alzheimer&apos s disease pathology Animal Model Apoptosis Behavioral Biological Assay Caenorhabditis elegans Characteristics Childhood Chromosome abnormality Chromosomes Clustered Regularly Interspaced Short Palindromic Repeats Code Consumption Defect Development Down Syndrome Early Onset Alzheimer Disease Epilepsy Essential Genes Etiology Frequencies GATA1 gene Gene Dosage Genes Genetic Health Human Individual Inherited Intellectual functioning disability Knock-in Lead Letters MAP Kinase Gene Methaqualone Modeling Molecular Mus Muscle Muscle function Mutate Natural regeneration Nematoda Nerve Degeneration Nervous system structure Neurons Orthologous Gene Pathway interactions Persons Phenotype Physiological Physiology Play Process Proteins RNA Interference Research Research Personnel Risk Role SYNJ1 gene Series Severities Sleep disturbances Speed Synapses Testing Time Transgenic Organisms axon guidance base behavioral study cholinergic cholinergic neuron congenital heart disorder cost dosage human stem cells improved in vivo in vivo Model induced pluripotent stem cell innovation insight knock-down leukemia motor disorder mouse model mutant neurobehavioral novel novel therapeutics overexpression partial trisomy 21 relating to nervous system screening stem cell model theories therapeutic target tool

项目摘要

PROJECT SUMMARY/ABSTRACT Down syndrome (DS) is the most common genetic cause of intellectual disability. For decades, we have known that an extra copy of the 21st chromosome (Hsa21) causes the broad array of physiological and developmental phenotypes associated with DS. However, we are far behind on addressing two critical questions in DS research: 1) Which Hsa21 genes cause phenotypes when overexpressed in DS, and conversely, 2) Which Hsa21 genes might be targeted to improve phenotypes in DS. Researchers using mouse models have discovered the role that a handful of Hsa21 genes play in certain phenotypes when overexpressed, such as Alzheimer’s pathology (APP) and leukemia (GATA1). The vast majority of Hsa21 genes, however, have not been studied in detail due to the impracticality of studying over 200 genes on Hsa21. Studying each of the Hsa21 genes using mouse models is time-consuming and costly. Instead, we propose to systematically study individual Hsa21 gene orthologs using the efficient model Caenorhabditis elegans. Recently, we found that, excluding 48 keratin genes, C. elegans has orthologs for over half of remaining Hsa21 genes, 51 of which are highly-conserved. Through mutant and RNAi analysis of the 51 orthologs, we found that 14 are essential genes and 10 are required for neural and/or muscular function, three of which had not previously been studied. To probe how overexpression (OE) of individual Hsa21 genes contributes to phenotypes relevant to DS, we will carry out complementary aims. For Aim 1, we will systematically test which of the 51 orthologs cause OE phenotypes one-by-one. For Aim 2, using a transgenic worm that carries extra copies of all 51 Hsa21 orthologs, we will systematically test which of the Hsa21 orthologs may be knocked down to reduce OE phenotypes. For OE phenotypes that we are able to suppress by gene knock down, we will also investigate the underlying cellular-molecular mechanistic bases with convenient approaches unique to C. elegans. Our project will leverage cutting-edge transgenic and gene knockdown tools, as well as take advantage of our lab’s expertise in high-throughput quantitative phenotyping. By identifying Hsa21 genes that cause phenotypes when overexpressed in C. elegans, this study will spotlight genes to prioritize for further study using mouse and human stem-cell models of DS. The set of Hsa21 OE strains produced will be shared freely around the world to establish C. elegans as the first mechanistic in vivo model to conveniently study consequences of Hsa21 gene overexpression. Insights gained from this study will highlight novel and uncharacterized molecular pathways as potential therapeutic targets for improving health in those with DS.

项目摘要/摘要唐氏综合症（DS）是智力残疾的最常见遗传原因。几十年来，我们知道第21染色体（HSA21）的额外副本引起了广泛的生理和与DS相关的发育表型。但是，我们远远落后于两个关键 DS研究中的问题：1）HSA21基因在DS中过表达时会引起表型，并且相反，2）HSA21基因可能被靶向以改善DS中的表型。研究人员使用鼠标模型已经发现了少数HSA21基因在某些表型中发挥的作用过表达的，例如阿尔茨海默氏病（APP）和白血病（GATA1）。 HSA21的绝大多数然而，由于在HSA21上研究了200多个基因的不切实际性，因此尚未详细研究基因。使用小鼠模型研究每个HSA21基因都是耗时且昂贵的。相反，我们建议使用有效的秀丽隐杆线虫模型，系统地研究单个HSA21基因直系同源物。最近，我们发现，除了48个角蛋白基因外，秀丽隐杆线虫具有剩余HSA21以上以上的直系同源物基因，其中51个是高度保存的。通过对51个直系同源物的突变和RNAi分析，我们发现 14是必不可少的基因，神经和/或肌肉功能需要10个，其中三个没有以前是研究。探测单个HSA21基因的过表达（OE）有助于与DS相关的表型，我们将执行完整的目标。对于AIM 1，我们将系统地测试哪个在51个直系同源物中，导致OE表型一对一。对于AIM 2，使用带有额外的转基因蠕虫所有51个HSA21直系同源物的副本，我们将系统地测试哪些HSA21直系同源物可能被敲击下降以减少OE表型。对于我们能够通过基因敲击抑制的OE表型，我们将还以方便的方法来研究潜在的细胞分子机械碱秀丽隐杆线。我们的项目将利用尖端的转基因和基因敲低工具，并采用我们实验室在高通量定量表型方面的专业知识的优势。通过识别HSA21基因在秀丽隐杆线虫中过表达时会引起表型，该研究将聚焦基因以进一步排序使用DS的小鼠和人类干细胞模型进行研究。产生的HSA21 OE菌株将共享在世界各地自由建立秀丽隐杆线虫作为方便研究的首个体内机械模型 HSA21基因过表达的后果。从这项研究中获得的见解将重点介绍小说和未表征的分子途径是改善DS患者健康的潜在治疗靶标。