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

聚集体中的特异性蛋白质可诊断大多数神经退行性疾病，包括阿尔茨海默病， 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。 微生物，并可能成为各种年龄进展性疾病的基础。通过定义聚集蛋白质组， 几个小鼠和人类组织，我们已经确定了一个非常大的部分聚集体成分， 在不同器官中常见，并在所有部位随年龄增长而增加。我们已经测试了几十个"共享" 通过RNAi敲低C.并发现大多数 有助于汇总过程-这表明存在一个汇总应计的准途径。更好地 了解这个有点有序的粘附过程，我们开发了改进的试剂，协议和分析 用于聚集体邻居的点击化学交联的工具。因此我们构建了聚合连接体， 这表明，少量的枢纽连接器介导了大聚集体的聚结， 否则将被分离成可能抵抗正常清除机制消化的大聚集体。 这里提出的策略是比较小鼠，随着年龄的增长，聚集的数量和组成， 大脑心脏和血清小鼠的基因型不同（野生型，或其BRI-A β 42转基因同窝出生的小鼠， 否则与野生型小鼠同基因，但由于"种子"的过度表达而随着年龄的增长形成脑淀粉样蛋白 肽"A β 1 - 42），以及饮食和药物干预。我们提出三个具体目标如下： 目标1。定义野生型小鼠随年龄增长（4 - 24岁）的心脏、脑和血清的聚集和总蛋白质组 月），±饮食和药物干预。在这里，我们将定义公共聚合组件， 衰老，由两种组织共享，部分反映在血清中发现的聚集体。我们预计， 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 通过先前显示可降低聚集或AD发病率的NSAID相关药物改善。 目标2.定义BRI-A β 42转基因小鼠心脏、大脑和血清的聚集体和总蛋白质组， 年龄（4 - 24个月），±饮食和药物干预。目标和程序与目标1相似， 增加记忆力和葡萄糖耐量测试。我们预期BRI-A β 42小鼠的脑聚集加速。 目标3。分析和追求衰老和/或疾病的候选生物标志物。我们将进行两两比较 正常饮食的对照组与每个干预组，以确定候选蛋白质和不同的PTM 年龄、组或基因型之间的差异> 2倍，FDR <0.01。我们将寻求的特点，单独或共同， 可用于预测年龄、淀粉样变性、记忆丧失、葡萄糖耐受不良或心脏的总负荷， 大脑血清聚集体具有特别的实用性，因为它们可以被非侵入性地评估。微差 在两种组织中观察到的特征，如果通过RD、阿司匹林和PNR 502改善，但通过WD和BRI加重。 A β 42（脑中）将通过交联相互作用组分析和通过shRNA敲低的功能测试来追踪。

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.