Project 1: Identifying genes and Pathways that impact Tau Toxicity in FTD

项目摘要/摘要 进行性核上性瘫痪(PSP)和皮质基底膜变性(CBD)是帕金森病 尸检中主要的tau病理改变。一种常见的微管相关非重组单倍型 染色体17q21上的蛋白tau基因(MAPT)基因座增加了PSP和CBD的风险，最近我们的 全基因组关联努力发现了额外基因/座位(STX6、EIF2AK3、SOS1、KIF13B、 和MOBP/Appoptosin)。此外，其他尚未检测到的基因可能与PSP的易感性有关 和CBD。本研究旨在解决外显子组和整体内与疾病相关的遗传变异。 来自PSP和CBD患者的基因组序列数据，以确定病理后果和 这些以tau病理为特征的复杂神经退行性疾病的潜在机制如下 确定潜在的治疗靶点。对于目标1，我们将分析一个由600多个数据组成的独特数据集 来自病理证实的PSP患者和350个外显子的2400个外显子和全基因组序列 来自CBD患者。这些数据将与通过阿尔茨海默氏症获得的5000个对照外显子进行比较 单核苷酸变异(SNV)分析的测序项目。在目标2中，我们将扩展我们对此的分析 对结构变异(SV)和拷贝数变异(CNV)进行分类和具体分析 PSP和CBD使用一系列分析软件程序，我们已经进行了广泛的测试以实现 对于每种类型的变化都具有最佳的灵敏度。对于目标3，我们将使用可用的外显子组和全基因组 序列数据，以评估tau毒性和病理与核心C的遗传变异的关联，通过 评估不同脑区的tau负荷和小胶质细胞增多症作为神经细胞丢失的替代指标。正在进行中 研究表明，使用定量病理方法可以帮助区分PSP的亚型。为了达到目标 4，我们将确定已确定的显著相关变异/基因的影响。我们将检测mRNAs 表达，RNA-Seq数据，体外功能研究，以表征观察到的突变对tau的影响 聚集和微管组装，以及这些基因/变异对tau蛋白水平和 死后脑组织中的异构体。综上所述，全外显子组和全基因组的结合 来自经病理证实的高度表型患者的序列数据，并有深入的分析计划 关注SNV、CNV、SV和数量性状，为新基因的发现提供了独特的机会。 识别新的紧张症基因是更好地理解 这组疾病背后的病理机制，可能有助于识别这些疾病的预后生物标志物 破坏性的疾病。

PROJECT SUMMARY/ABSTRACT Progressive supranuclear palsy (PSP) and corticobasal degeneration (CBD) are Parkinsonian disorders with predominant tau pathology at autopsy. A common non-recombining haplotype at the microtubule associated protein tau gene (MAPT) locus on chromosome 17q21 increases the risk of PSP and CBD and recently our genome-wide association efforts identified variation in additional genes/loci (STX6, EIF2AK3, SOS1, KIF13B, and MOBP/Appoptosin). In addition, other genes, as yet undetected, likely contribute to susceptibility to PSP and CBD. This study aims to resolve the disease-associated genetic variation within the exome and whole genome sequence data from PSP and CBD patients, to determine the pathological consequences and mechanisms underlying these complex neurodegenerative diseases characterized by tau pathology, thus identifying potential therapeutic targets. For Aim 1, we will analyze a unique dataset consisting of over 600 exomes and 2400 whole genome sequences derived from pathology-confirmed PSP patients and 350 exomes from CBD patients. These data will be compared to the 5000 control exomes available through the Alzheimer's sequencing project for single nucleotide variant (SNV) analysis. In Aim 2, we will expand our analysis of this cohort and specifically analyze structural variants (SV) and copy number variations (CNV) that contribute to PSP and CBD using a range of analytical software programs which we have extensively tested to achieve optimal sensitivity for each type of variation. For Aim 3, we will use the available exome and whole genome sequence data to assess the association of genetic variation on tau toxicity and pathology with Core C, by assessing tau burden in different brain regions and microgliosis as a surrogate of neuronal cell loss. On-going efforts have shown that using quantitative pathologic measures can help distinguish subtypes of PSP. For Aim 4, we will determine the effect of significantly associated variants/genes identified. We will examine mRNA expression, RNA-Seq data, in vitro functional studies to characterize the effect of the observed mutation on tau aggregation and microtubule assembly, and the effect of these genes/variants on tau protein levels and isoforms in postmortem brain tissue. In summary, the combination of whole exome and whole genome sequence data from pathologically-confirmed and highly-phenotyped patients with an in-depth analytical plan focusing on SNV, CNV, SV and quantitative traits, provides a unique opportunity for novel gene discovery. Identifying novel genes for tauopathies is a critical step towards a better understanding of the pathomechanisms underlying this group of disorders and may help identify prognostic biomarkers for these devastating disorders.