Regulation of the Autophagy Process in Organismal Aging

项目摘要 自噬是一种主要的细胞再循环过程，通过该过程，胞质货物在细胞内被隔离和降解。 溶酶体这个多步骤的过程在发育、疾病和衰老中起着重要作用。直接联系 自噬和衰老之间的联系存在于多种保守的长寿模式中;这些长寿的动物是 被认为以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牢固地建立了自噬和线虫寿命之间的联系 C.通过显示自噬基因是所有保守长寿的长寿所必需的， 范式测试此外，我们未发表的结果表明，自噬在功能上与 长寿C.我们迄今为止分析的线虫突变体， 通常在长寿突变体的组织中出现诱导，但在野生型动物中随时间下降。我们和 其他人也表明，自噬的几种转录和翻译后调节因子在 老化，这表明自噬随着时间的推移受到复杂的调节。 虽然共同确立了自噬在C. elegans这些研究 没有解决哪些组织特异性功能自噬可能控制影响生物体健康的问题。 此外，目前还不清楚自噬在长寿突变体中以及在基因突变过程中是如何在时间和空间上进行调节的。 正常老化。此应用程序的目标是通过使用以下组合来解决这些知识差距 遗传、生物化学和行为分析，主要是在C.也存在于哺乳动物系统中。 具体来说，在目标1中，我们将使用定量PCR和靶向蛋白质组学来表征自噬是如何在细胞中表达的。 在C.线虫和鼠组织。此外，我们将使用SILAC耦合 蛋白质组学来测量C.线虫组织在目标2中，我们 分析自噬在C. elegans healthspan，并分析对健康的影响-并 以时间和空间控制的方式过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节剂， 自噬，包括自噬货物识别的重要因素。 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 神经变性了解自噬的调节和保守的机制， 自噬影响多细胞生物的衰老，如C.秀丽线虫很可能提供新的重要见解 这不仅有助于衰老，还可能有助于开发治疗此类与年龄有关的疾病的方法。

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 Moreover, our unpublished results indicate that autophagy is functionally relevant for longevity in all major tissues of the long-lived C. elegans mutants we have analyzed so far, and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.