Identifying protective omics profiles in centenarians and translating these into preventive and therapeutic strategies

评估跨多个分析平台的蛋白质测量的准确性。我们的NIA资助 UH2 AG064704项目“确定百岁老人的保护性组学特征，并将其转化为 预防和治疗战略“，是一项分阶段的尿毒症/尿毒症3项目。我们目前处于UH2阶段，即 我们实现了具体的里程碑，为开展UH3阶段奠定了基础并为其提供了便利。其中 作为UH2的主要里程碑，我们正在招募和招募1,400名百岁老人及其后代，我们是 收集他们的表型数据、血液和粪便样本。在UH3阶段，我们将产生多组学 这些样本的特征，并将它们与人类的极端长寿表型相关联。其他专业 UH2阶段的里程碑包括规划生成组学数据和开发分析 我们将在项目的UH3阶段使用的管道。我们在以下方面取得了实质性进展 对于大多数组学数据的登记、数据管理和计划，我们也意识到了 使用准确的数据生成平台，该平台与其他人类研究使用的平台很好地协调 极端长寿，如长寿家庭研究和长寿联盟。在我们的父应用程序中， 我们建议使用SOMAscan技术来产生血清蛋白质组学。另类蛋白质组学 这些方法包括标记质谱仪和Olink Explore平台。这三大主流 技术在所需的样本准备、人类蛋白质组的覆盖范围、 蛋白质丰度评估的准确性、蛋白质检测的特异性及其性能还没有 进行了全面的比较。在这项行政补充请求中，我们建议 生成数据，为选择用于UH3阶段的最佳蛋白质组平台提供信息 项目。我们有机会与其他关于人类极端长寿的研究联合起来，以比较质量 光谱、SOMAscan和Olink平台使用了精心设计的尖峰实验。我们建议两个 明确的目标。具体目标1：生成250个样本的基于SomaLogic的蛋白质组图谱， 代表一个对照条件的三个副本，以及6个池，包括13个不同的添加的8个不同的蛋白质 丰度，对于总共3+3𝑥6𝑥13=237.为保证质量，我们将再增加13个空白样品 控制力。具体目标2：对使用 SOMAscan、Olink和质谱学技术。将使用经过清理和标准化的数据来分析 基于剂量的蛋白质丰度轨迹，以检测不同池中变化的蛋白质并 评估SOMAscan、Olink和MS检测蛋白质的灵敏度和特异度 技术。Somascan技术与替代技术产生的数据的比较 将提供关键信息来指导选择用于UH3阶段的最佳蛋白质组技术 父项目的。

Assessing accuracy of protein measurements across multiple analytical platforms. Our NIA funded project, UH2AG064704 “Identifying protective omics profiles in centenarians and translating these into preventive and therapeutic strategies”, is a phased UH2/UH3 project. We are currently in the UH2 phase, in which we achieve specific milestones to set the stage for and facilitate the conduct of the UH3 phase. Among the major UH2 milestones, we are recruiting and enrolling 1,400 centenarians and their offspring, and we are collecting their phenotype data, blood and fecal samples. In the UH3 phase, we will generate multi-omics profiles of those samples and correlate them with extreme human longevity phenotypes. Other major milestones of the UH2 phase include planning for the generation of omics data and developing analyses pipelines that we will use in the UH3 phase of the project. We have made substantial progress toward enrollment, data management, and plans for the majority of omics data and we also realized the importance of using an accurate data generating platform that is well harmonized with those used by other studies of human extreme longevity such as the Long Life Family Study and the Longevity Consortium. In our parent application, we proposed to use the SOMAscan technology to generate serum proteomics. Alternative proteomics approaches include labelled mass spectrometry and the Olink Explore platforms. These three mainstream technologies have pros and cons in terms of required sample preparation, coverage of the human proteome, accuracy of protein abundance assessment, and specificity of proteins detection and their performance has not been compared in a comprehensive way. In this request for an administrative supplement we propose to generate data that will inform the choice of the best proteomic platform to be used in the UH3 phase of the project. We have the opportunity to join forces with other studies of human extreme longevity to compare mass spectrometry, SOMAscan and Olink platforms using a well-designed spike-in experiment. We propose two specific aims. Specific Aim 1: To generate SOMAlogic-based proteomic profiles of 250 samples that represent triplicates of one control condition, and 6 pools including 8 different proteins spiked at 13 different abundance, for a total of 3 + 3𝑥6𝑥13 = 237 samples. We will add an additional 13 blank samples for quality control. Specific Aim 2: To conduct quality control analysis of the proteomic profiles generated using the SOMAscan, Olink and Mass-spectrometry technologies. Cleaned and normalized data will be used to analyze the dosage-based trajectories of protein abundance to detect proteins that change in the different pools and to estimate sensitivity and specificity of protein detection of the SOMAscan, Olink and mass-spectrometry technologies. The comparison of data generated with the Somascan technology versus alternative technology will provide critical information to guide the selection of the best proteomic technology to use in the UH3 phase of the parent project.