Validation and optimization of epigenetic clocks

项目概要/摘要 寻找和验证延长人类健康寿命的分子靶点一直具有挑战性，因为大多数 临床生物标志物既不充分机械化也不接近衰老的基本机制 作为指标。为了应对开发衰老生物标志物的挑战，我们将主要关注 表观遗传改变，因为通过这项工作，DNA 甲基化 (DNA) 很可能会作为一种 有效的生物标志物可供临床应用。该提案的总体目标是 a) 实现伟大的目标 基于 DNAm 的衰老生物标志物（称为表观遗传时钟）对人类干预研究的前景 b) 增进他们对机制的理解。该提案建立在我们积极的研究计划的基础上 围绕基于 DNAm 的衰老生物标志物及其与细胞衰老标志物的关系。我们 和其他人已经证明，现有的 DNAm 生物标志物原型可以预测寿命和 许多与年龄有关的病症。我们已经确定一些 DNAm 生物标志物与生活方式干预有关 现有的 DNAm 生物标志物已被用于抗衰老干预措施的人体临床试验。与 这些概念验证研究完成后，我们进入了下一阶段，这些生物标志物需要被 优化以跟踪人类研究干预措施的有效性。尽管人们普遍承认 DNAm 生物标志物非常稳健，它们对技术变化仍然敏感。为了尽量减少此类 虚假变异，我们将优化工作流程中的所有组件，从人体样本的收集到 最后的分析点。 在我们之前的规划拨款中，我们的研究人员网络设计了一项评估 DNAm 效用的研究 基于人类纵向队列研究中衰老的生物标志物。在这些计划的基础上，我们将测试是否 DNAm 生物标志物是人类健康寿命中基本衰老过程的指标。 纵向队列研究、遗传学研究和介入研究。我们将把 DNAm 生物标志物与 细胞衰老标志物和一系列衰老的临床生物标志物，以推进机制 见解。我们将评估一组生物标志物是否更能预测健康寿命/多种健康状况 结果而不是单一标记。我们将优化用于人类离体研究的 DNAm 生物标志物 用于测试对各种抗衰老或促衰老干预措施的反应。由此产生的优化系统还 作为一个潜在的高通量药物筛选系统，它将具有可控的优势 潜在的混杂因素，并且是人类起源的。总的来说，这些调查将导致 (a) DNAm生物标志物在人体介入治疗中应用的整个工作流程的优化 研究和 (b) 使用背景陈述，充分描述每个生物标志物的目的和用途。

Project Summary/Abstract It has been challenging to find and validate molecular targets for extending human healthspan because most clinical biomarkers are neither sufficiently mechanistic nor proximal to fundamental mechanisms of aging to serve as indicators. To address the challenge of developing biomarkers of aging, we will primarily focus on epigenetic alterations because it is highly likely that with this work DNA methylation (DNA) will come out as a valid biomarker ready for clinical application. The overarching goals of this proposal are a) to realize the great promise of DNAm based biomarkers of aging (known as epigenetic clocks) for human interventional studies and b) to advance their mechanistic understanding. This proposal builds on our active research program surrounding DNAm based biomarkers of aging and their relationship to markers of cellular senescence. We and others have demonstrated that existing prototypes of DNAm biomarkers are predictive of lifespan and many age-related conditions. We have established that some DNAm biomarkers relate to lifestyle interventions and existing DNAm biomarkers are already being used in human clinical trials of anti-aging interventions. With these proof-of-concept studies completed, we enter the next phase where these biomarkers need to be optimized to track the effectiveness of interventions in human studies. Although it is widely acknowledged that DNAm biomarkers are remarkably robust, they remain sensitive to technical variation. To minimize such spurious variation, we will optimize all components within the workflow, from collection of human samples to the final point of analysis. In our previous planning grant, our network of researchers designed a study for evaluating the utility of DNAm based biomarkers of aging in human longitudinal cohort studies. Building on these plans, we will test whether DNAm biomarkers are indicators of a fundamental aging process underlying healthspan by carrying out human longitudinal cohort studies, genetic studies, and interventional studies. We will relate DNAm biomarkers to markers of cellular senescence and a battery of clinical biomarkers of aging in order to advance mechanistic insights. We will evaluate whether a panel of biomarkers is more predictive of healthspan/multiple health outcomes as opposed to a single marker. We will optimize DNAm biomarkers for use in human ex vivo studies for testing responsiveness to various anti-aging or pro-aging interventions. The resulting optimized system also serves as a potential high-throughput drug screening system, which will have the advantage of being controlled for potential confounding factors and is of human origin. Collectively, these investigations will result in (a) the optimization of the entire workflow process for the application of DNAm biomarkers in human interventional studies and (b) context-of-use statements that fully describe the purpose and use of each biomarker.