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

项目总结 巨噬细胞自噬(称为自噬)是一种主要的细胞循环过程，通过这种过程胞浆物质被 被隔离成双膜小泡或自噬小体(即自噬的早期步骤)，这是 随后与溶酶体融合以确保货物降解(即自噬的后期步骤)。这个综合体， 多步骤过程在生物发育和年龄相关疾病中起着关键作用，并有许多直接的 自噬和衰老之间存在联系，包括需要多种保守的长寿范例 延长寿命的自噬基因；目前的范例表明，这种长寿的动物 自食性周转是一种有益的方式，但其根本机制仍然难以捉摸。 我们在前一个资金周期中的研究提供了对这一关系的更深层次的理解 在线虫的自噬和衰老之间。首先，我们使用细胞学标记的工作已经 表明在不同组织中具有不同轨迹的自噬后期步骤与年龄相关的下降。 此外，我们还观察到在长寿突变体中组织特异性自噬的不同调节。 时尚。其次，我们在线虫体内发现了特定自噬基因的意外长寿作用 神经元，表明可能不同于溶酶体降解的非规范功能，这是一种新兴的 自噬研究领域的概念，这一概念尚未在老龄化背景下进行调查。最后，我们 进行了无偏见的遗传和生化筛查，以确定新的候选自噬调节器和 受体，并与我们的合作者一起发现了一种小分子化合物，它可以增加两者的自噬 在哺乳动物细胞和线虫体内，并延长寿命。 我们之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进行深入的测试。 具体地说，我们将使用基因和生化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衰老和长寿有关的一个限制步骤 (目标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作用。 在长寿和疾病模型中(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，如神经退行性变。 了解自噬的调节以及自噬影响衰老的保守机制 像线虫这样的多细胞生物不仅可能为衰老提供新的重要见解，而且可能 还有助于开发治疗此类与年龄相关的疾病的方法，包括神经退行性疾病。

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.