Novel misfolded proteins in ADRD: proteomics, genetics, and clinical-pathological correlations

ADRD 中的新型错误折叠蛋白：蛋白质组学、遗传学和临床病理相关性

• 批准号: 9905466
• 负责人: PETER T. NELSON
• 金额: $ 19.13万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-15 至 2022-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9905466
• 关键词: Age; Alzheimer' s disease pathology; Alzheimer' s disease related dementia; Amygdaloid structure; Amyloid beta-Protein; Animal Model; Automobile Driving; Autopsy; Biochemical; Biochemistry; Biological Markers; Brain; Brain Diseases; Candidate Disease Gene; Cessation of life; Chromosome Mapping; Clinical; Clinical Data; Core Facility; Correlation Studies; Data; Databases; Dementia; Detergents; Development; Diagnostic; Disease; Education; Elderly; Electrospray Ionization; Freezing; Funding Mechanisms; Future; Genes; Genetic; Genetic study; Genie; Genomics; Goals; Hand; Heterogeneity; Human; Image; Immunohistochemistry; Impaired cognition; Impairment; Individual; Kentucky; Knowledge; Liquid Chromatography; Mass Spectrum Analysis; Mutation; Neurodegenerative Disorders; Neurofibrillary Tangles; Pathogenicity; Pathologic; Pathology; Persons; Process; Proteins; Proteome; Proteomics; Public Health; Reporting; Research; Research Project Grants; Sampling; Seasons; Series; Severities; Statistical Data Interpretation; Statistical Models; TDP-43 aggregation; Testing; Therapeutic; Universities; University resources; Urea; Work; aged; aging brain; alpha synuclein; base; biobank; cerebrovascular pathology; clinical Diagnosis; cohort; demented; endophenotype; frontotemporal lobar dementia-amyotrophic lateral sclerosis; genetic risk factor; genetic variant; insight; large scale data; misfolded protein; multidisciplinary; neuropathology; novel; polypeptide; protein TDP-43; protein misfolding; sex; synuclein; tandem mass spectrometry; tau Proteins

The heterogeneous pathologies of Alzheimer's disease and related dementias (ADRD) are still incompletely understood. There is no perfect animal model to study these diseases. Four known aberrantly misfolded proteins are commonly detected via neuropathologic examination of brains with ADRD: Aβ, Tau, TDP-43, and/or α-Synuclein. Our preliminary data indicate that there are more pathogenic protein species awaiting identification. Characterization of these proteins may be relevant to the development of both diagnostic and therapeutic strategies for ADRD. The goals of this research project are to identify previously uncharacterized, misfolded, and aberrantly processed proteins in ADRD; to resolve gene variants that modulate the severity and heterogeneity of dementia through the novel misfolded proteins; and, to perform clinical-pathological correlation, focusing on these novel proteins, to identify new disease-associated pathologic biomarkers. We will leverage the resources of the University of Kentucky AD Center (UK-ADC) biobank and the University of Kentucky Proteomics Core Facility, to execute the following Specific Aims: Specific Aim 1. Analyse detergent-insoluble protein extracts from human amygdala (snap-frozen at autopsy) to investigate whether as yet uncharacterized misfolded proteins are detectable in ADRD. Polypeptides from the detergent-insoluble, urea-soluble protein fractions of amygdala will be interrogated using mass spectrometry. We have on-hand amygdalae from 40 UK-ADC subjects which incorporate a spectrum of clinical and pathologic features. Immunohistochemistry and biochemical studies will be performed to determine the best candidate proteins for additional studies (Aims 2 and 3). Specific Aim 2. Construct a robust and harmonized database with exonic sequencing to localize genetic regions associated with novel misfolded proteins in ADRD. We will test the association between dementia and rare, exonic (`mis-sense') genetic variants in genes that encode candidate misfolded proteins (Aim 1). Genetics data augmented with rich neuropathologic endophenotypes and longitudinal clinical data will enable insights into novel mechanisms that drive dementia. Large-scale datasets (NACC, ADGC, ADSP) will be aggregated and harmonized to test the genetic drivers of clinical and neuropathology-based endophenotypes focusing on novel candidate genes. Candidate target genes have already been identified. Specific Aim 3. Establish the clinical-pathologic correlation for the novel misfolded proteinopathies in ADRD. We will test the association between the potential novel proteinopathies (from Aim 1) with cognitive impairment, factoring in known markers Aβ, tau, α-Synuclein, and TDP-43, as well as cerebrovascular pathologies. We have 370 samples available for analyses (both sexes) from the UK-ADC, which should be statistically powered to test our hypothesis that the presence and severity of novel misfolding protein pathologies are associated with cognitive impairment.

阿尔茨海默病和相关痴呆 (ADRD) 的异质病理学仍然存在差异 不完全理解。目前还没有完美的动物模型来研究这些疾病。四种已知异常 错误折叠的蛋白质通常通过 ADRD 大脑的神经病理学检查来检测：Aβ、Tau、 TDP-43 和/或 α-突触核蛋白。我们的初步数据表明存在更多的致病蛋白 等待鉴定的物种。这些蛋白质的表征可能与 ADRD 的诊断和治疗策略。该研究项目的目标是确定 ADRD 中先前未表征、错误折叠和异常加工的蛋白质；解决基因变异 通过新型错误折叠蛋白调节痴呆症的严重程度和异质性；并且，到 进行临床病理关联，重点关注这些新蛋白质，以确定新的疾病相关 病理生物标志物。我们将利用肯塔基大学 AD 中心 (UK-ADC) 的资源 生物银行和肯塔基大学蛋白质组学核心设施，以实现以下具体目标： 具体目标 1. 分析人杏仁核中不溶于洗涤剂的蛋白质提取物（速冻 尸检时）以研究在 ADRD 中是否可以检测到尚未表征的错误折叠蛋白。 来自杏仁核不溶于去污剂、可溶于尿素的蛋白质部分的多肽将被研究 使用质谱法。我们现有 40 个 UK-ADC 受试者的杏仁核，其中包含 临床和病理特征谱。免疫组织化学和生化研究将 以确定用于进一步研究的最佳候选蛋白质（目标 2 和 3）。 具体目标 2. 构建一个强大且统一的数据库，通过外显子测序进行本地化 与 ADRD 中新型错误折叠蛋白相关的遗传区域。我们将测试之间的关联 痴呆症和编码候选错误折叠蛋白的基因中罕见的外显子（“错义”）遗传变异 （目标 1）。遗传学数据加上丰富的神经病理内表型和纵向临床数据将 深入了解导致痴呆症的新机制。大型数据集（NACC、ADGC、ADSP）将 进行汇总和协调，以测试基于临床和神经病理学的遗传驱动因素 内表型侧重于新的候选基因。候选靶基因已经被确定。 具体目标 3. 建立新型错误折叠的临床病理相关性 ADRD 中的蛋白质病。我们将测试潜在的新型蛋白质病之间的关联（来自 目标 1) 认知障碍，考虑已知标记物 Aβ、tau、α-突触核蛋白和 TDP-43，以及 脑血管病变。我们有来自 UK-ADC 的 370 个样本可供分析（男女）， 这应该有统计能力来检验我们的假设，即新颖性的存在和严重性 错误折叠的蛋白质病理与认知障碍有关。