An innovative proof-of-concept approach to identify age-modulating drugs capable of reversing inflammation and re-setting the epigenetic clock

摘要 为了确定有可能延迟年龄开始和进展的药物和/或干预措施- 相关的病理学，临床前研究已经使用了动物模型，其中可以延长寿命。 准确评估。在过去的十年中，已经确定了几种优秀的候选治疗方法; 这些包括mTOR抑制剂（如雷帕霉素）、衰老清除剂、阿卡波糖等。然而，翻译 从动物模型到人类的抗衰老结果一直是一个挑战。例如，测量 因此，有必要确定和验证可以为人类服务的参数， 作为健康衰老的替代标志。此外，年龄调节药物的疗效往往只是 在老年人中，功能缺陷已经很明显。因此，测试潜在的抗- 衰老治疗应该在一组高龄的人类受试者中进行; 在这种“高危”人群中，给药的风险甚至更为严重。为了避免这些限制，我们 建议在人类受试者中开发和验证一种创新的、微创的、具有成本效益的检测方法 年龄在65-95岁之间。推定的抗衰老试验药物将局部应用于皮肤的离散区域， 在受试者前臂上的皮肤上，同时将“仅载体”（对照）平行施加到相对的手臂。因此， 每个受试者将作为其自身对照。“试验”和安慰剂中心的药效学结局 将在男性和女性中进行比较，因为年龄调节剂的作用通常在 性别确定局部应用的治疗剂的功效需要稳健的衰老生物标志物， 可在皮肤中测量，但可推广至完整的生物体。为此，我们选择 使用两个独立的参数，这两个参数已知会随着局部（在单个组织中）老化而改变。 和系统性：i）DNA甲基化模式或“表观遗传时钟”和ii）与以下相关的生物标志物： 炎症为了验证所提出的技术，我们选择了一种药物，雷帕霉素（RAPA）， 已经显示出调节啮齿动物的衰老，并且已经测试了安全的全身应用， 人类[1-4]。6个月治疗期后，将在每个部位产生抽吸水泡， 收集用于表观基因组分析的皮肤细胞（水泡瓣）（Aim 1）和用于表观基因组分析的间质（水泡）液（Aim 2）。 用于测量细胞因子和其他炎症调节因子（目的2）。一旦得到验证，我们的方法可以 用于安全有效地筛选药理学药物在人体中的年龄调节潜力， 确定那些药物，保证大，昂贵的人体临床试验与全身药物管理。

ABSTRACT In order to identify drugs and/or interventions with the potential to delay the onset and progression of age- associated pathologies, preclinical studies have used animal models where extension of lifespan could be accurately assessed. Several outstanding candidate therapeutics have been identified over the last decade; these include mTOR inhibitors (like rapamycin), senolytic agents, acarbose, and others. However, translating anti-aging results from animal models into humans has been challenging. For example, measuring extension of lifespan would be impractical in humans, so it is necessary to identify and validate parameters that can serve as surrogate markers of healthy aging. Moreover, the efficacy of age-modulating drugs is often only demonstrable with older individuals where functional deficits are already evident. Thus, testing of potential anti- aging therapeutics should be done in a cohort of human subjects of an advanced age; issues of safe administration would be even more acute in this “at risk” population. To circumvent these limitations, we propose to develop and validate an innovative, minimally invasive, cost-effective assay in human subjects ranging from 65-95 years old. The putative anti-aging test drug will be applied topically to a discrete area of skin on the subject’s forearm while a “vehicle only” (control) is applied in parallel to the opposite arm. Thus, each subject will serve as his/her own control. The pharmacodynamic outcomes at the “test” and placebo sites will be compared in both males and females since the effects of age-modulating agents often differ between the sexes. Ascertaining the efficacy of topically-applied therapeutics requires robust biomarkers of aging that can be measured in skin, but will be generalizable to the intact organism. Towards this end, we have opted to use two independent parameters that are known to be modified with aging both locally (in individual tissues) and systemically: i) the DNA methylation pattern or “epigenetic clock” and ii) biomarkers associated with inflammation. To validate the proposed technique, we have chosen a drug, rapamycin (RAPA), which has already been shown to modulate aging in rodents and which has been tested for safe systemic application in humans [1-4]. After a 6 month treatment phase, suction blisters will be generated at each site to allow collection of skin cells (blister flap) for use in the epigenomic analysis (Aim 1) and interstitial (blister) fluid for use in measuring cytokines and other inflammatory regulators (Aim 2). Once validated, our approach can be used to safely and efficiently screen the age-modulating potential of pharmacological agents in humans to identify those agents that warrant large, expensive human clinical trials with systemic agent administration.