Inference of Common Pathways Underlying Neurodegeneration & Other Age-Progressive Diseases

神经退行性病变常见途径的推断

• 批准号: 10170207
• 负责人: Srinivas Ayyadevara
• 金额: $ 37.5万
• 依托单位: UNIV OF ARKANSAS FOR MED SCIS
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-30 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10170207
• 关键词: Adult; Age; Age-Months; Aging; Alzheimer' s Disease; Alzheimer' s disease brain; Amyloid; Amyloid beta-42; Amyloidosis; Aspirin; Brain; C57BL/6N Mouse; Cachexia; Caenorhabditis elegans; Caloric Restriction; Cardiovascular Diseases; Cerebrum; Characteristics; Chemistry; Computer software; Data; Diabetes Mellitus; Diagnostic; Diet; Digestion; Disease; Disease model; Elements; Exclusion Criteria; Failure; Fatty acid glycerol esters; Freezing; Functional disorder; Genotype; Glucose Intolerance; Glucose tolerance test; Goals; Heart; Human; Huntington Disease; Incidence; Individual; Insulin Resistance; Intervention; Investigation; Kidney; Kidney Failure; Mediating; Mediator of activation protein; Memory; Memory Loss; Methods; Modeling; Mus; Muscle; Muscular Atrophy; Myocardium; Nature; Nerve Degeneration; Neurodegenerative Disorders; Non-Steroidal Anti-Inflammatory Agents; Organ; Organism; Parkinson Disease; Pathway interactions; Peptides; Pharmaceutical Preparations; Population; Post-Translational Protein Processing; Procedures; Process; Progressive Disease; Proteins; Proteome; Proteomics; Protocols documentation; RNA Interference; Reagent; Reporting; Risk Factors; Role; Seeds; Serum; Site; Skeletal Muscle; Sucrose; Testing; Tissues; Transgenic Mice; Transgenic Organisms; Wild Type Mouse; adhesion process; age group; age related; aged; analytical tool; base; candidate marker; cardiovascular risk factor; cerebral amyloidosis; connectome; crosslink; dietary; dietary restriction; effective therapy; functional decline; glucose tolerance; group intervention; human tissue; improved; knock-down; novel strategies; overexpression; prophylactic; prospective; protein aggregation; protein protein interaction; proteostasis; response; small hairpin RNA; western diet; young adult

Specific proteins within aggregates are diagnostic for most neurodegenerative diseases, including Alzheimer’s, Parkinson’s, and Huntington’s diseases. Protein aggregation, however, is a basic feature of aging in numerous organisms, and may underlie a variety of age-progressive diseases. By defining the aggregate proteomes from several mouse and human tissues, we have identified a very large fraction of aggregate components that are common across diverse organs, and increase with age in all sites. We have tested several dozen “shared” components for functional roles in aggregation by RNAi knockdown in C. elegans, and found that the majority contribute to the aggregation process – suggesting that there is a quasi-pathway for aggregate accrual. To better understand this somewhat orderly adhesion process, we developed improved reagents, protocols and analytic tools for click-chemistry crosslinking of aggregate neighbors. We thus constructed aggregate connectomes, which revealed that a small number of hub connectors mediate coalescence of large aggregates, which would otherwise be isolated, into mega-aggregates that may resist digestion by the normal clearance mechanisms. The strategy proposed here is to compare mice, as they age, for aggregate quantity and composition in cerebrum, heart, and serum. The mice will differ in genotype (wild-type, or their BRI-Aβ42 transgenic littermates, otherwise isogenic with the wild-type mice, but forming brain amyloid with age due to overexpression of a “seed peptide” Aβ1–42), and also with respect to dietary and drug interventions. We propose 3 Specific Aims, as follows: Aim 1. Define aggregate and total proteomes of heart, brain, and serum from wild-type mice as they age (4 – 24 months), ± dietary and drug interventions. Here we will define common aggregate components that accrue with aging, shared by two tissues and partially reflected by aggregates found in serum. We expect many of these proteins or their post-translational modifications to respond to restricted diet (RD) and western diet (WD), and be ameliorated by NSAID-related drugs that were previously shown to reduce aggregation or AD incidence. Aim 2. Define aggregate and total proteomes of heart, brain, and serum from BRI-Aβ42 transgenic mice as they age (4 – 24 months), ± dietary and drug interventions. Goals and procedures parallel those for Aim 1, with the addition of memory and glucose-tolerance tests. We expect accelerated brain aggregation in BRI-Aβ42 mice. Aim 3. Analysis and pursuit of candidate biomarkers of aging and/or disease. We will make pairwise comparisons of controls on normal diet with each intervention group, to identify candidate proteins and PTMs that differ between ages, groups or genotypes by >2-fold, with FDR<0.01. We will seek features that, individually or jointly, can be used to predict age, amyloidosis, memory loss, glucose intolerance, or aggregate burden in heart or cerebrum. Serum aggregates have particular utility since they can be assessed noninvasively. Age-differential features observed in both tissues, if ameliorated by RD, aspirin and PNR502, but exacerbated by WD and BRI- Aβ42 (in brain), will be pursued by cross-linking interactome analysis and functional testing by shRNA knockdown.

聚集体中的特定蛋白质对大多数神经退行性疾病都有诊断作用，包括阿尔茨海默氏症， 帕金森氏症和亨廷顿氏症。然而，蛋白质聚集是许多人衰老的基本特征 生物体，并可能是各种年龄进行性疾病的基础。通过定义来自的聚合蛋白质组 几种小鼠和人体组织，我们已经鉴定出很大一部分聚集体成分是 在不同的器官中普遍存在，并且在所有部位随着年龄的增长而增加。我们已经测试了几十个“共享” 通过RNAi敲除线虫中的聚集功能成分，发现大多数 对聚合过程做出贡献--这表明存在一条聚合应计利润的准路径。为了更好地 了解了这种有序的黏附过程，我们开发了改进的试剂、方案和分析 用于聚合邻居的点击化学交联的工具。因此，我们构建了聚合连接， 它揭示了少量的集线器连接器调节大集合体的结合，这将 否则，将被分离成巨型聚集体，可能会抵抗正常清除机制的消化。 这里提出的策略是比较随着年龄增长的小鼠的聚集数量和组成 大脑、心脏和血清。这些小鼠的基因型将不同(野生型，或它们的BRI-Aβ42转基因窝产仔， 在其他方面与野生型小鼠基因相同，但由于一种“种子”的过度表达，随着年龄的增长而形成脑淀粉样蛋白 肽“Aβ1-42)，也涉及饮食和药物干预。我们提出三个具体目标，具体如下： 目的1.确定野生型小鼠心脏、脑和血清随年龄增长的聚合和总蛋白质组(4-24 月)，±饮食和药物干预。在这里我们将定义公共聚合组件，这些组件由 衰老，由两个组织共享，部分反映在血清中的聚集体中。我们预计会有很多这样的 蛋白质或其翻译后修饰，以响应限制饮食(RD)和西方饮食(WD)，以及 以前被证明可以减少聚集或AD发病率的非甾体抗炎药相关药物可以改善病情。 目的2.确定BRI-Aβ42转基因小鼠心、脑和血清的聚合和总蛋白质组 年龄(4-24个月)，±饮食和药物干预。目标和程序与目标1类似，具有 增加了记忆和葡萄糖耐量测试。我们预计BRI-Aβ42小鼠的脑聚集加速。 目的3.分析和寻找衰老和/或疾病的候选生物标志物。我们将进行两两比较 对每个干预组的正常饮食进行对照，以确定不同的候选蛋白质和PTM 年龄、群体或基因型之间按&gt；2倍，用FDR&lt；0.01。我们将单独或联合寻找以下特征： 可用于预测年龄、淀粉样变性、记忆力减退、糖耐量异常或心脏或 大脑。血清聚集物具有特殊的用途，因为它们可以被无创性地评估。年龄差异 在两种组织中观察到的特征，如果用RD、阿司匹林和PNR502改善，但被WD和BRI加重- β42(在大脑中)将通过交联相互作用组分析和shRNA敲除功能测试来寻找。