Regulation of lifespan

寿命调节

• 批准号: 9473004
• 负责人: Kurt W Runge
• 金额: $ 32.49万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-01 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9473004
• 关键词: Affect; Aging; Aging-Related Process; Animal Model; Autophagocytosis; Binding; Biological Assay; Caloric Restriction; Cell Survival; Chemicals; Complex; Cyclin-Dependent Kinases; Cyclins; Data; Development; Disease; Fission Yeast; Genetic; Genetic screening method; Genomic approach; Goals; Health; Homologous Gene; Human; Intervention; Link; Longevity; Mammals; Mediating; Molecular; Mutate; Neurons; Nutrient; Organelles; Orthologous Gene; Pathway interactions; Phosphorylation; Phosphorylation Site; Phosphotransferases; Positioning Attribute; Process; Proteins; Regulation; Role; Signal Pathway; Signal Transduction Pathway; Societies; Testing; Therapeutic Intervention; Variant; Yeasts; aging population; analog; analytical tool; design; detection of nutrient; dietary restriction; innovation; novel; novel strategies; protein aggregate; therapeutic candidate; therapeutic target

Project Summary The aging population and coincident rise in associated diseases demand a better understanding of the basic mechanisms of aging to design appropriate interventions and prolong human health. Elucidating the molecular mechanisms of the processes that slow aging, such as lifespan extension by caloric or dietary restriction, would provide prime candidates for therapeutic intervention. Model organisms have allowed major advances in defining aging mechanisms due to their exceptionally powerful genetics and analytical tools, and because these processes are evolutionarily conserved from humans to yeasts. We discovered a new pathway in Schizosaccharomyces pombe that both regulates autophagy and extends lifespan. Autophagy is a process that degrades and recycles proteins and organelles, and has been linked to lifespan and diseases associated with human aging. We found that both autophagy and lifespan are regulated through the cyclin-dependent kinase Pef1, an ortholog of human Cdk5. Pef1 acts with its cyclin Clg1 to limit lifespan and autophagy, and is opposed by the effector kinase Cek1 that extends lifespan and increases autophagy levels. The Clg1-Pef1- Cek1 pathway acts independently of TOR, the only other pathway known to regulate both lifespan and autophagy, but how these processes are controlled by the Pef1 pathway is unknown. Aim 1 will test the hypothesis that the Pef1 pathway senses nutrients to control lifespan and autophagy. This aim will be significant for testing the linkage between autophagy and lifespan extension by caloric or dietary restriction. Establishing the mechanism by which the Pef1 pathway regulates lifespan and autophagy requires knowing the downstream effectors, and Aim 2 describes a novel chemical genomics approach to identify Pef1 targets. We modified the Pef1 kinase to allow selective binding of ATP analogs that tag kinase substrates to allow their isolation, and used the Pef1 variant to identify the substrates. We identified substrates with predicted roles in autophagy and lifespan, as well as substrates that we hypothesize mediate Pef1 control through signaling pathways. Aim 2 will validate the substrates and identify substrates of the effector kinase Cek1. Aim 3 will determine how the phosphorylation of Pef1 and Cek1 substrates affects specific processes in lifespan and autophagy control, and will also use high-throughput functional characterization of substrates to reveal signaling pathways regulated by the Pef1 pathway. The strong functional homology between Pef1 and human Cdk5 indicates that the Clg1-Pef1-Cek1 pathway, its substrates and functions will be conserved in humans, which we will also test in Aim 3. The results of these proposed studies will provide a mechanistic framework for a new, conserved lifespan and autophagy-regulating pathway with important ramifications for healthy human aging.

项目摘要 人口老龄化和相关疾病的同步上升需要更好地了解基本的 衰老机制，以设计适当的干预措施，延长人类健康。阐明分子 减缓衰老过程的机制，如通过热量或饮食限制延长寿命， 将为治疗干预提供主要候选人。模式生物已经取得了重大进展 在定义衰老机制方面，由于他们非常强大的遗传学和分析工具， 这些过程从人类到酵母在进化上是保守的。我们发现了一条新的途径， 粟酒裂殖酵母既调节自噬又延长寿命。自噬是一个过程 它降解和分解蛋白质和细胞器，并与寿命和相关疾病有关。 与人类衰老的关系我们发现，自噬和寿命都是通过细胞周期蛋白依赖性调节的。 激酶Pef 1，人Cdk 5的直系同源物。Pef 1与其细胞周期蛋白Clg 1一起限制寿命和自噬， 与之相反的是效应激酶Cek 1，它可以延长寿命并增加自噬水平。Clg1-Pef1- Cek 1通路独立于TOR发挥作用，TOR是已知调节寿命和细胞周期的唯一其他通路。 自噬，但这些过程是如何控制的Pef 1途径是未知的。目标1将测试 Pef 1通路感知营养以控制寿命和自噬的假说。这一目标将是 这对于测试自噬与通过热量或饮食限制延长寿命之间的联系具有重要意义。 建立Pef 1通路调节寿命和自噬的机制需要了解 的下游效应，和目标2描述了一种新的化学基因组学方法，以确定Pef 1的目标。 我们修饰了Pef 1激酶，使其能够选择性结合标记激酶底物的ATP类似物， 分离，并使用Pef 1变体来鉴定底物。我们确定了底物与预测的作用， 自噬和寿命，以及我们假设通过信号传导介导Pef 1控制的底物 路径。目的2将验证底物并鉴定效应激酶Cek 1的底物。目标3将 确定Pef 1和Cek 1底物的磷酸化如何影响寿命中的特定过程， 自噬控制，并将使用高通量的功能表征底物，以揭示 由Pef 1途径调节的信号通路。Pef 1与人的强功能同源性 Cdk 5表明Clg 1-Pef 1-Cek 1通路、其底物和功能在人类中是保守的， 我们也将在Aim 3中进行测试。这些研究的结果将提供一个机制框架 对于一个新的，保守的寿命和自噬调节途径， 人类衰老