Validation and optimization of epigenetic clocks

项目摘要/摘要 寻找和验证延长人类健康寿命的分子靶点一直是一个挑战，因为大多数 临床生物标记物既不具有足够的机械性，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记物的挑战，我们将主要集中在 表观遗传学改变，因为很有可能随着这项工作，DNA甲基化(DNA)将成为一种 可供临床应用的有效生物标志物。这项提议的首要目标是：a)实现 基于dNaM的衰老生物标志物(称为表观遗传学时钟)在人类干预研究中的前景 以及b)推进他们的机械性理解。这项建议建立在我们积极研究计划的基础上。 周围基于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生物标记物原型可以预测寿命和 许多与年龄有关的疾病。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生物标记物已经用于抗衰老干预的人体临床试验。使用 这些概念验证研究完成后，我们进入下一个阶段，这些生物标记物需要 经过优化以跟踪人体研究中干预措施的有效性。尽管人们普遍认为 DNaM生物标记物非常健壮，它们对技术变化仍然敏感。要最大限度地减少这种 虚假变异，我们将优化工作流中的所有组件，从人类样本的收集到 最后一点分析。 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评估dNaM的效用的研究。 人类纵向队列研究中衰老的生物标记物。在这些计划的基础上，我们将测试 DNaM生物标记物是健康寿命潜在的基本衰老过程的指示器，通过进行人类 纵向队列研究、遗传学研究和干预研究。我们将dNaM生物标记物与 细胞衰老标志物和一系列临床衰老生物标志物的研究进展 洞察力。我们将评估一组生物标志物是否能更好地预测健康状况 结果，而不是单一的标记。我们将优化dNaM生物标记物用于人体体外研究 用于测试对各种抗衰老或促衰老干预措施的反应性。由此产生的优化系统还 作为一种潜在的高通量药物筛选系统，它将具有受控的优势 潜在的混杂因素，而且是人类起源的。总而言之，这些调查将导致(A) DNaM生物标志物在人类介入治疗中应用的整个工作流程优化 研究和(B)充分描述每个生物标记物的目的和用途的使用背景说明。

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.