Novel genetic modifiers of C9orf72 and Tau toxicity

C9orf72 和 Tau 毒性的新型遗传修饰剂

• 批准号: 10084641
• 负责人: John David Fryer
• 金额: $ 403.72万
• 依托单位: MAYO CLINIC ARIZONA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-15 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10084641
• 关键词: Novel; genetic; modifiers; C9orf72; Tau

Hexanucleotide (GGGGCC) repeat expansions in C9orf72 are the leading genetic cause of frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS). In this proposal, we seek to identify new therapeutic targets by using a genetically diverse mouse model of C9-FTD/ALS. In this study design, we will be using adeno-associated virus (AAV) to drive mutant C9-repeat expression (AAV-149R) in a panel of Collaborative Cross (CC) recombinant inbred mice, as well as Diversity Outbred (DO) mice produced from random repeated outcrossing of CC strains. Collectively, the CC and DO mouse populations offer high mapping resolution and broad allelic diversity, carrying 45 million SNPs and structural variants. This provides a unique opportunity to discover new genes that regulate C9-repeat toxicity in vivo. These modifiers will be compared to a parallel study of CC/DO mice that will be injected with AAV-TauP301L to determine modifiers of tau toxicity. In Aim 1 we will use CC mice to identify strains that are sensitive or resistant to C9 or tau toxicity while in Aim 2 we will use the DO mice to finely map these potential modifiers. In Aim 3 we will perform single- cell RNAseq studies from AAV-149R and AAV-TauP301L mice at stages of pre-symptomatic, as well as moderate and severe disease to determine the evolution of the brain response to these pathologies at the individual cell level. Also in Aim 3, we will perform single-nucleus RNAseq on postmortem human tissue from healthy controls, c9-ALS, c9-FTD, and FTLD-TauP301L from frontal cortex. Top hits from these datasets will be used to prioritize candidate genes/loci from Aims 1 and 2, and followed up with validation studies on postmortem tissue at the histological and biochemical level. At the conclusion of this comprehensive study, we hope to have identified numerous modifiers of both diseases that can be followed up for more in depth functional studies by our laboratories or the broader scientific community. It is likely that some of these genetic modifiers could also impact other neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, etc. and could be followed up by targeted testing in those disease models.

六核苷酸（GGGGCC）在C9ORF72中重复扩张是主要的遗传原因 额颞痴呆（FTD）和肌萎缩性侧索硬化症（ALS）。在此提案中，我们试图确定 通过使用C9-FTD/ALS的遗传多样的小鼠模型，新的治疗靶标。在这项研究设计中，我们 将使用腺相关病毒（AAV）来驱动突变体C9重复表达（AAV-149R） 协作杂交（CC）重组近交小鼠，以及从 随机重复的CC菌株过渡。 CC和DO鼠标共同提供高度 映射分辨率和广泛的等位基量多样性，具有4500万个SNP和结构变体。这提供了 在体内发现调节C9重复毒性的新基因的独特机会。这些修饰符将是 与将注入AAV-TAUP301L的CC/DO小鼠的平行研究相比 tau毒性。在AIM 1中，我们将使用CC小鼠识别对C9或TAU毒性敏感或抗性的菌株 在AIM 2中，我们将使用DO MICE来精细绘制这些潜在的修饰符。在AIM 3中，我们将执行单一 来自AAV-149R和AAV-TAUP301L小鼠的细胞RNASEQ在症状前的阶段以及 中度和重度疾病以确定大脑对这些病理的反应的演变 单个细胞水平。同样在AIM 3中，我们将对从 来自额叶皮层的健康对照，C9-ALS，C9-FTD和FTLD-TAUP301L。这些数据集中的最高点击将是 用于从AIMS 1和2中优先考虑候选基因/基因座，然后进行验证研究 在组织学和生化水平的事后组织。在这项综合研究的结论中，我们 希望已经确定了这两种疾病的许多修饰符 我们的实验室或更广泛的科学界的功能研究。其中一些遗传可能 修饰符还可能影响其他神经退行性疾病，例如阿尔茨海默氏病，帕金森氏病 疾病等，可以在这些疾病模型中进行针对性测试。