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.

C9 orf 72中的六核苷酸（GGGGCC）重复扩增是导致C9 orf 72突变的主要遗传原因。 额颞叶痴呆（FTD）和肌萎缩侧索硬化（ALS）。在这一建议中，我们寻求确定 通过使用C9-FTD/ALS的遗传多样性小鼠模型的新治疗靶标。在这项研究设计中，我们 将使用腺相关病毒（AAV）驱动突变C9-重复表达（AAV-149 R）在一组 协作杂交（CC）重组近交系小鼠，以及由以下产生的多样性远交（DO）小鼠： CC品系的随机重复异交。总的来说，CC和DO小鼠群体提供了高的 该基因组的定位分辨率和广泛的等位基因多样性，携带4500万个SNP和结构变体。这提供了 这是一个发现体内调节C9重复毒性的新基因的独特机会。这些修改器将 与将注射AAV-TauP 301 L的CC/DO小鼠的平行研究相比， tau毒性。在目标1中，我们将使用CC小鼠来鉴定对C9或tau毒性敏感或具有抗性的品系 而在目标2中，我们将使用DO小鼠来精细地映射这些潜在的修饰符。在目标3中，我们将执行单一- 来自症状前阶段的AAV-149 R和AAV-TauP 301 L小鼠的细胞RNAseq研究，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。同样在目标3中，我们将对来自人的死后人体组织进行单核RNAseq。 健康对照、c9-ALS、c9-FTD和来自额叶皮质的FTLD-TauP 301 L。这些数据集的热门点击率将是 用于对目标1和2的候选基因/位点进行优先排序，并跟进以下验证研究： 在组织学和生物化学水平上对死后组织进行了研究。在这项全面研究结束时，我们 我希望已经确定了这两种疾病的许多修饰符，可以进行更深入的随访 我们的实验室或更广泛的科学界的功能研究。很可能这些基因 修饰剂也可以影响其他神经退行性疾病，如阿尔茨海默病，帕金森病， 疾病等，并且可以通过在这些疾病模型中进行有针对性的测试来跟进。