Regulation of the Autophagy Process in Organismal Aging

项目总结 自噬是一种主要的细胞循环过程，通过这种过程，细胞内的货物被隔离并在 溶酶体。这一多步骤的过程在发育、疾病和衰老过程中发挥着重要作用。直接通航 自噬和衰老之间存在于多种保守的长寿范式中；这种长寿动物是 被认为以一种有益的方式诱导自噬，但潜在的机制仍然难以捉摸。 我们和其他人的研究已经在线虫的自噬和寿命之间建立了牢固的联系 线虫通过证明自噬基因是所有保守的长寿所必需的 测试的范例。此外，我们未发表的结果表明，自噬在功能上与 到目前为止，我们已经分析过的长寿线虫突变体的所有主要组织中的寿命以及自噬 通常在长寿突变体的组织中被诱导，但在野生型动物中随着时间的推移而下降。我们和 其他研究也表明，自噬的几个转录和翻译后调节因子在 随着时间的推移，自噬会受到复杂的调控。 这些研究在线虫的多个组织中共同确立了自噬的关键作用 没有说明自噬可能控制哪些组织特异性功能，从而影响机体的健康寿命。 此外，尚不清楚自噬在时间和空间上如何在长寿突变体和自噬期间受到调节。 正常衰老。此应用程序的目标是通过组合使用以下各项来解决这些知识差距 遗传、生化和行为分析主要在线虫中进行，但也在哺乳动物系统中进行。 具体地说，在目标1中，我们将使用定量聚合酶链式反应和靶向蛋白质组学来表征自噬是如何 在线虫和小鼠组织中，衰老过程是受调控的。此外，我们将使用SILAC耦合 用蛋白质组学方法测量线虫组织中精选自噬货物的降解率。在目标2中，我们将 分析自噬在线虫健康寿命中的组织特异性作用，并分析对健康的影响-和 以时间和空间受控的方式过度表达关键自噬调节基因的寿命。 最后，在目标3中，我们将使用遗传和生化筛选方法来寻找新的调控因子 自噬，包括识别自噬货物的重要因素。 自噬在许多疾病中起着关键作用，包括与年龄相关的疾病，如 神经退行性变。了解自噬的调节和通过其保守的机制 自噬影响线虫等多细胞生物的衰老可能提供新的重要见解 不仅会导致衰老，而且还可能有助于开发此类与年龄相关的疾病的治疗方法。

PROJECT SUMMARY Autophagy is a major cellular recycling process by which cytosolic cargo is sequestered and degraded in the lysosome. This multi-step process plays important roles in development, disease, and aging. Direct links between autophagy and aging exist in multiple conserved longevity paradigms; such long-lived animals are thought to induce autophagy in a beneficial manner, yet the underlying mechanisms remain elusive. Ours and others' research have firmly established links between autophagy and longevity in the nematode C. elegans by showing that autophagy genes are required for the long lifespan of all conserved longevity paradigms tested. Moreover, our unpublished results indicate that autophagy is functionally relevant for longevity in all major tissues of the long-lived C. elegans mutants we have analyzed so far, and autophagy generally appears induced in tissues of long-lived mutants, but declines over time in wild-type animals. We and others have also shown that several transcriptional and post-translational regulators of autophagy play roles in aging, suggesting that autophagy is subject to complex regulation over time. While collectively establishing a critical role for autophagy in multiple tissues of C. elegans, these studies did not address which tissue-specific functions autophagy may control that affect organismal healthspan. Moreover, it is unclear how autophagy is temporally and spatially regulated in long-lived mutants and during normal aging. The goal of this application is to address these gaps in knowledge by using a combination of genetic, biochemical and behavioral assays primarily in C. elegans, but also in mammalian systems. Specifically, in Aim 1, we will use quantitative PCR and targeted proteomics to characterize how the autophagy process is regulated during aging in C. elegans and murine tissues. Moreover, we will use SILAC-coupled proteomics to measure degradation rates of select autophagy cargos in C. elegans tissues. In Aim 2, we will analyze tissue-specific roles for autophagy in C. elegans healthspan, and analyze the effects on health- and lifespan of overexpressing key autophagy-regulatory genes in a temporal and spatial-controlled manner. Finally, in Aim 3, we will use genetic and biochemical screening approaches to search for new regulators of autophagy, including factors important for autophagic cargo recognition. Autophagy plays critical roles in many diseases, including age-related disorders such as neurodegeneration. Understanding the regulation of autophagy and the conserved mechanisms by which autophagy affect aging in multicellular organisms like C. elegans are likely to provide new important insights not only into aging but may also help develop treatments for such age-related diseases.