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，因为大多数人都认为， 临床生物标志物既不充分机械，也不接近老化的基本机制， 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，因为它是非常有可能的，与这项工作的DNA甲基化（DNA）将出来，作为一个 有效的生物标志物准备用于临床应用。该提案的总体目标是：（a）实现 基于DNAm的衰老生物标志物（称为表观遗传时钟）有望用于人类干预研究 和B）推进他们的机械理解。这项建议建立在我们积极的研究计划之上 周围的基于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， 许多与年龄有关的问题。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生物标志物已经被用于抗衰老干预的人体临床试验。与 这些概念验证研究完成后，我们进入了下一个阶段，这些生物标志物需要 优化以跟踪人类研究中干预措施的有效性。尽管人们普遍认为， DNAm生物标志物非常稳健，它们对技术变化保持敏感。尽量减少这种 虚假的变化，我们将优化工作流程中的所有组件，从人类样本的收集到 分析的最后一点。 在我们之前的计划拨款中，我们的研究人员网络设计了一项研究，用于评估DNAm的效用。 基于人类纵向队列研究中的衰老生物标志物。根据这些计划，我们将测试 DNAm生物标志物是通过进行人类健康评估的基础衰老过程的指标。 纵向队列研究、遗传学研究和干预性研究。我们将把DNA生物标志物与 细胞衰老的标志物和一系列衰老的临床生物标志物，以推进衰老的机制。 见解.我们将评估一组生物标志物是否更能预测健康span/多种健康 而不是一个单一的标志。我们将优化DNAm生物标志物用于人体离体研究 用于测试对各种抗衰老或促衰老干预的反应性。所得到的优化系统还 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，并且是人源性的。总的来说，这些调查将导致（a） 在人类介入治疗中应用DNAm生物标志物的整个工作流程的优化 研究和（B）充分描述每种生物标志物的目的和用途的使用背景声明。

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.