Regulation of tau expression in Alzheimer disease and aging

阿尔茨海默病和衰老中 tau 表达的调节

基本信息

批准号：
9315684
负责人：
John Fonda Crary
金额：
$ 81.26万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-07-15 至 2021-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9315684
关键词：
17q21 3&apos Untranslated Regions Aging Alleles Alzheimer&apos s Disease Amino Acids Amyloid beta-Protein Astrocytes Attention Autopsy Biochemical Brain Brain Diseases Chromosomes Classification Clinical Clinical Research Code Computer Analysis DNA sequencing Data Diagnosis Diagnostic Disease Equilibrium Event Failure Foundations Future Genes Genetic Genetic Risk Genetic Transcription Genetic study Genotype Goals Haplotypes Impairment Individual Light Link MAPT gene Maps Medicine Messenger RNA MicroRNAs Molecular Molecular Genetics Mutation Nerve Degeneration Neurofibrillary Tangles Nucleotides Parkinson Disease Pathogenesis Pathology Phenotype Play Post-Transcriptional Regulation Post-Translational Protein Processing Production Prognostic Marker Progressive Supranuclear Palsy Protein Isoforms Proteins Public Health RNA RNA Splicing RNA-Binding Proteins Regulation Research Resolution Risk Role Scientist Stratification System Tauopathies Techniques Testing Time Transcript Traumatic Brain Injury Validation Variant age related brain tissue burden of illness chronic traumatic encephalopathy corticobasal degeneration density diagnostic biomarker disease classification disorder subtype genome-wide genome-wide analysis improved in vivo innovation interest multimodality novel novel diagnostics novel therapeutics pleiotropism prion-like risk variant single molecule tau Proteins tau expression

项目摘要

The 17q21.31 MAPT locus is associated with Alzheimer's disease (AD) and sporadic primary tauopathies in the absence of a tau gene coding region mutation, but the causative variants remain unknown. Our long-term goal is to understand the pathogenesis of tauopathies and pave the way towards novel diagnostics and therapeutics. The objective here is to use new diagnostic criteria to better stratify subjects for a molecular genetic study of tauopathy. Using high-resolution fine mapping and long-read single molecule real time (SMRT) DNA sequencing that employs phospholinked nucleotides to produce very long reads (>70,000 bp) to overcome structural complexity at the locus and pinpoint functional alleles. Further, we will use long-read mRNA isoform sequencing (IsoSeq) to assess mRNA levels, splicing and alternative 3' untranslated regions (UTRs) that might influence post-transcriptional regulation, including RNA binding proteins and microRNAs. We will validate our findings using biochemical, histopathological and cellular techniques. Our central hypothesis is that distinct subhaplotypes confer risk for different MAPT-associated diseases through dysregulation of the expression of toxic tau species leading to unique higher order degenerative phenotypes. Our rationale is that understanding these changes will be crucial for diagnostic stratification and further clinical and mechanistic studies. We will test our hypothesis by pursuing the following specific aims: Aim 1. To identify functional risk alleles associated with tauopathy, we will generate a high density genetic map of the 17q21.31 locus using high-coverage genotyping and targeted SMRT sequencing to characterize the locus and correlate novel variation with AD, primary age-related tauopathy, progressive supranuclear palsy, corticobasal degeneration, Parkinson disease and other tauopathies to pinpoint functional risk alleles; Aim 2. To discover and validate novel mRNA splicing events in tauopathy, we will perform a genome wide computational analysis using brains from subjects with AD and other tauopathies to discover novel splicing events in tau and other genes as well as changes in splicing factors then validate these findings using biochemical, histopathological and cellular approaches; and Aim 3. To uncover novel post-transcriptional regulators of tauopathy, we will discover and validate both protein and RNA post-transcriptional regulators of tau and tauopathy and characterize their networks and validate these changes using biochemical, histopathological and cellular approaches. This contribution will be significant because it will provide a foundation for further mechanistic studies that will elucidate the drivers of disease and pave the way for future clinical studies. This proposal is innovative because by studying AD in the context of the tauopathies there is the possibility that it will lead to a harmonized classification system.

17q21.31 MAPT 位点与阿尔茨海默病 (AD) 和不存在 tau 基因编码区突变的散发性原发性 tau 病相关，但致病变异仍然未知。我们的长期目标是了解 tau蛋白病的发病机制，并为新的诊断和治疗方法铺平道路。这里的目标是使用新的诊断标准更好地对受试者进行分层，以进行 tau 蛋白病的分子遗传学研究。使用高分辨率精细定位和长读长单分子实时 (SMRT) DNA 测序，利用磷酸化核苷酸产生超长读长 (>70,000 bp)，以克服基因座的结构复杂性并精确定位功能等位基因。此外，我们将使用长读长 mRNA 同工型测序 (IsoSeq) 来评估可能影响转录后调控的 mRNA 水平、剪接和替代 3' 非翻译区 (UTR)，包括 RNA 结合蛋白和 microRNA。我们将使用生化、组织病理学和细胞技术验证我们的发现。我们的中心假设是，不同的亚单倍型通过有毒 tau 物种表达失调导致独特的高阶退行性表型，从而赋予不同 MAPT 相关疾病的风险。我们的理由是，了解这些变化对于诊断分层以及进一步的临床和机制研究至关重要。我们将通过追求以下具体目标来检验我们的假设：目标 1. 为了识别与 tau 蛋白病相关的功能性风险等位基因，我们将使用高覆盖率基因分型和靶向 SMRT 测序来生成 17q21.31 位点的高密度遗传图谱，以表征基因座并将新变异与 AD、原发性年龄相关 tau 病、进行性核上性麻痹、皮质基底节变性、帕金森病和其他 tau 病联系起来查明功能风险等位基因；目标 2. 为了发现和验证 tau 蛋白病中新的 mRNA 剪接事件，我们将使用 AD 和其他 tau 蛋白病受试者的大脑进行全基因组计算分析，以发现 tau 和其他基因中的新剪接事件以及剪接因子的变化，然后进行验证这些发现采用生化、组织病理学和细胞方法；目标 3. 为了发现 tau 蛋白病的新型转录后调节因子，我们将发现并验证 tau 蛋白和 tau 蛋白病的蛋白质和 RNA 转录后调节因子，并使用生化、组织病理学和细胞方法表征它们的网络并验证这些变化。这一贡献意义重大，因为它将为进一步的机制研究奠定基础，阐明疾病的驱动因素，并为未来的临床研究铺平道路。该提案具有创新性，因为通过在 tau 蛋白病的背景下研究 AD，有可能形成一个统一的分类系统。