Regulation of the Autophagy Pathway with Age and in Long-lived Animals

项目摘要 大自噬（Macroautophagy）（称为自噬）是一种主要的细胞再循环过程，通过该过程，胞质物质被细胞质吞噬。 隔离到双膜囊泡或自噬体中（即，自噬的早期步骤）， 随后与溶酶体融合以确保货物降解（即，自噬的后期步骤）。这个复杂的， 多步骤过程在生物体发育和与年龄相关的疾病中起着关键作用， 自噬和衰老之间存在联系，包括多种保守的长寿模式需要 自噬基因的寿命延长;目前的范例表明，这种长寿的动物诱导 自噬周转以有益的方式，但潜在的机制仍然难以捉摸。 我们在上一个资助周期的研究提供了对这种关系的更深入理解 在线虫C.优雅首先，我们使用细胞学标记的工作 表明在不同组织中，自噬的后期步骤与年龄相关的下降具有可变的轨迹。 此外，我们还观察到，在一个组织特异性突变体中， 时尚.其次，我们发现了C. elegans 神经元，表明可能与溶酶体降解不同的非典型功能， 这是自噬研究领域的一个概念，尚未在衰老的背景下进行研究。最后我们 进行了无偏见的遗传和生化筛选，以确定新的候选自噬调节剂， 受体，并与我们的合作者一起确定了一种小化合物，可以增加自噬， 哺乳动物细胞和C. elegans优雅and extends延长life. 我们先前的研究为我们提供了新的和具体的假设，我们的目标是在这次更新中深入测试。 具体来说，我们将使用遗传和生物化学方法的强大组合来询问融合-和/或 溶酶体降解是自噬过程中与生物体衰老和寿命相关的限制性步骤 (Aim 1）;我们将询问神经元自噬基因是否通过细胞非自主方式影响衰老（目标2）;以及， 最后，我们将询问新的候选自噬受体和自噬调节分子是否具有 在寿命和疾病模型中的作用（目标3）。 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。 了解自噬的调节和自噬影响衰老的保守机制， 多细胞生物如C.秀丽线虫可能提供新的重要见解，不仅对衰老， 还有助于开发治疗这些与年龄有关的疾病，包括神经退行性疾病的方法。

PROJECT SUMMARY Macroautophagy (referred to as autophagy) is a major cellular recycling process by which cytosolic material is sequestered into double-membrane vesicles or autophagosomes (i.e., early steps of autophagy), which subsequently fuse with lysosomes to ensure cargo degradation (i.e., late steps of autophagy). This complex, multi-step process plays key roles in organismal development and age-related diseases, and numerous direct links exist between autophagy and aging, including that multiple conserved longevity paradigms require autophagy genes for their lifespan extension; the current paradigm suggest that such long-lived animals induce autophagic turnover in a beneficial manner, yet the underlying mechanisms remain elusive. Our research in the previous funding cycle has provided a deeper understanding of the relationship between autophagy and aging in the nematode C. elegans. First, our work using cytological markers has indicated an age-related decline in late-steps of autophagy with variable trajectories in different tissues. Moreover, we have observed a differential regulation of autophagy in long-lived mutants in a tissue-specific fashion. Second, we have discovered unexpected longevity roles for specific autophagy genes in C. elegans neurons, indicating possibly non-canonical functions different from lysosomal degradation, an emerging concept in the autophagy research field which has yet to be investigated in the context of aging. Finally, we have conducted unbiased genetic and biochemical screens to identify new candidate autophagy regulators and receptors, and together with our collaborators identified a small compound that increase autophagy in both mammalian cells and in C. elegans and extends lifespan. Our prior studies provide us with new and specific hypotheses that we aim to test in depth in this renewal. Specifically, we will use a powerful combination of genetic and biochemical approaches to ask if fusion- and/or lysosomal degradation is a limiting step of the autophagy process in relation to organismal aging and longevity (Aim 1); we will ask if neuronal autophagy genes affect aging by cell non-autonomous means (Aim 2); and, finally, we will ask if new candidate autophagy receptors and autophagy-modulating molecules possess roles in longevity and in disease models (Aim 3). Autophagy plays critical roles in many disorders, including age-linked diseases 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 and may also help develop treatments for such age-related diseases, including neurodegenerative disorders.