Role of Clock-Modulating Small Molecules Against Aging

描述（申请人提供）：随着美国预期寿命和老年人口的增加，如何实现健康老龄化并延长健康寿命是一个具有深远社会影响的紧迫生物医学问题。控制我们健康的一个关键生物学机制是生物钟，这是一种内在的计时装置，可以对环境变化做出反应，并在整个24小时周期内协调身体功能。越来越多的证据表明，衰老和昼夜节律功能障碍之间存在很强的相关性，特别是昼夜节律的衰减，如睡眠片段化和体温和循环激素周期幅度的降低。我们最近通过高通量筛选确定了一组独特的时钟增强小分子，现在被称为CEM。为了研究时钟衰减在衰老中的潜在因果作用，我们将讨论以下假设： CEMs可以改善老化的生物钟和与年龄相关的代谢下降。我们专注于能量代谢，因为它受到生物钟的密切调节，而衰老与能量利用率的显著下降有关。提出了三个具体目标。 具体目标1：确定老年小鼠中CEMs的时钟机制。使用老年PER 2：：luc报告小鼠，我们将确定CEMs是否可以在组织和单细胞水平上增强生物发光节律。为了了解分子机制，我们将系统地描述衰老组织中的核心时钟循环，并剖析CEM介导的衰老时钟增强的转录和转录后机制。 具体目标2：阐明CEM在年龄相关代谢下降中的作用。我们将通过分子和生理学方法来确定CEMs是否可以增强自然衰老小鼠的能量代谢。为了确定CEM在老年小鼠能量稳态中的分子机制，我们将筛选响应CEM治疗的表达或活性改变的候选代谢调节因子，并研究CEM在老年小鼠中的代谢调节机制。 具体目标3：识别蜂窝网络和CEM的直接目标。为了鉴定老年小鼠对CEM的慢性和急性细胞反应，我们将使用来自用CEM治疗不同时期的老年小鼠的样品进行RNA-seq转录组分析。为了确定直接的目标，我们将进行化学蛋白质组学研究，涉及亲和下拉与生物素化的CEM衍生物。这些研究中特定细胞通路和蛋白质的抗衰老作用将在 通过药理学和遗传学方法进一步研究。 这些目标的成功完成将解决关于衰老过程中时钟衰减的因果作用的关键问题，并揭示CEM在延长健康寿命方面的令人兴奋的功效。

DESCRIPTION (provided by applicant): With the rising life expectancy and elderly population in the US, how to achieve healthy aging and extend health span is a pressing biomedical question with profound social ramifications. A pivotal biological mechanism governing our well-being is the circadian clock, the intrinsic timekeeping device that responds to environmental changes and coordinates bodily functions throughout the 24-h cycles. Accumulating evidence has demonstrated a strong correlation between aging and circadian dysfunctions, particularly attenuation of circadian rhythms such as sleep fragmentation and reduced amplitude of body temperature and circulating hormone cycles. We recently identified a unique group of Clock-Enhancing small Molecules, now dubbed as CEMs, via high-throughput screening. To investigate the potential causal role of clock attenuation in aging, we will address the hypothesis that CEMs can improve aged clocks and age-related metabolic decline. We focus on energy metabolism because it is closely regulated by the clock and aging is associated with significant decline in energy utilization. Three Specific Aims are proposed. Specific Aim 1: Determine the clock mechanism of CEMs in aged mice. Using aged PER2::luc reporter mice, we will determine whether CEMs can enhance the bioluminescence rhythm at tissue and single-cell levels. To understand the molecular mechanism, we will systematically characterize the core clock loops in aged tissues, and dissect transcriptional and posttranscriptional mechanisms underlying CEM-mediated enhancement of aged clocks. Specific Aim 2: Delineate the role of CEMs in age-related metabolic decline. We will determine whether CEMs can enhance energy metabolism in naturally aged mice by molecular and physiological approaches. To define the molecular mechanism of CEMs in energy homeostasis of aged mice, we will screen candidate metabolic regulators for altered expression or activity in response to CEM treatment, and investigate the metabolic regulatory mechanisms by CEMs in aged mice. Specific Aim 3: Identify the cellular networks and direct targets of CEMs. To identify both chronic and acute cellular responses to CEMs in aged mice, we will conduct RNA-seq transcriptome profiling using samples from aged mice treated with CEMs for varying periods. To identify direct targets, we will carry out chemoproteomic studies involving affinity pull-down with biotinylated CEM derivatives. Anti-aging roles of specific cellular pathways and proteins from these studies will be further investigated by pharmacological and genetic approaches. Successful completion of these Aims will address the critical question regarding a causal role of clock attenuation during aging and reveal an exciting efficacy of CEMs in prolonging health span.