Characterization of Conserved Protein Synthesis Aging Pathways

所有的人都会变老，尽管衰老的速度是高度可变的，很可能受到一个人的遗传组成的影响; 我们的细胞随着时间的推移积累损伤，这种损伤会导致疾病易感性增加， 对损伤的反应能力下降，感觉系统减少，以及许多其他有害变化。 虽然它们本身不一定致命，但它们会导致死亡风险大大增加。为了开始 在减少衰老影响的临床工作中，研究必须首先阐明引起衰老的生物化学相互作用。 这样的效果。本项目将鉴定和描述包含保守的长寿相关蛋白的通路 利用模式生物C.优雅线虫C. elegans是一个很好的模型， 由于其寿命短，细胞数量少，易于遗传操作和大部分（>80%）而老化 保守的基因。发育后蛋白质合成基因的表达减少， 寿命，甚至高达50%，但这种寿命增加的原因尚不清楚。最近有人 描述了如果在幼虫阶段给药，减少蛋白质合成会导致发育停滞。给定 这种联系，以及先前描述类似长寿相关拮抗多效性的多项研究，我们 寻求表征两种表型中涉及的蛋白质合成途径;这包括检测 假设这两种表型可能使用相同的途径，因为我们最近已经确定了这两个 这些状态共享应激抗性表型（增加的健康寿命的度量）。目标1将编制一份清单， 当蠕虫因蛋白质减少而停滞或长寿时，通过RNA-seq解除基因调控 合成.利用这个列表，和三个以前确定的基因，可以控制逮捕表型，我们将 使用GFP结合的启动子表征其时空表达模式以及它们如何相互关联 具有长寿、停滞和抗应激表型。最后，我们将确定是否拯救野生型 这些去调控基因的表达水平拯救了任何相同的表型，表明它们的重要性。 在回应中。目标2将确定以组织特异性方式减少蛋白质合成的影响 （皮下，肠，肌肉或神经元）使用组织特异性RNAi菌株，这将确定在哪里， 蠕虫的长寿途径正在发生，包括揭示如何信号和细胞间串扰 细胞类型之间的相互作用，赋予长寿，逮捕和应激抗性。目标2的第二部分 包括从我们的时空分析和组织特异性研究中整合信息， 发现在赋予这些生存促进表型中重要的基因，以确定它们的 在细胞或组织水平上的重要性。这些蛋白质合成基因的描述，特别是在 与它们的生理效应、发现的途径以及与停滞状态和压力的相关性有关 将在增强寿命和如何赋予寿命之间建立更好的联系 以便能够最终治疗与衰老有关的疾病。

All humans age, though the rate of aging is highly variable and likely influenced by one's genetic composition; our cells accumulate damage over time, and this damage can lead to increased disease susceptibility, decreased ability to respond to injury, reduction in sensory systems, among many other detrimental changes. While not necessarily deadly on their own, they lead to a greatly increased risk of death. In order to begin the clinical work in reducing the effects of aging, research must first elucidate the biochemical interactions causing such effects. This project will identify and describe pathways containing conserved longevity-related protein synthesis genes using the model organism, C. elegans. The nematode C. elegans is an excellent model for aging due to its short lifespan, small number of cells, ease of genetic manipulation, and large fraction (>80%) of conserved genes. The reduced expression of protein synthesis genes post-developmentally increases worm lifespan, even up to 50% higher, but the reasons for this lifespan increase are unknown. It has recently been described that reducing protein synthesis cause developmental arrest if given during the larval stage. Given this connection, and multiple previous studies describing similar longevity-related antagonistic pleiotropy, we seek to characterize the protein synthesis pathways involved in both phenotypes; this includes testing the hypothesis that these two phenotypes may use the same pathway as we have recently determined both of these states share a stress resistance phenotype (a metric of increased healthspan). Aim 1 will compile a list of deregulated genes via RNA-seq when worms undergo arrest or longevity in response to reduced protein synthesis. Using this list, and three previously-identified genes that can control the arrest phenotype, we will characterize their spatiotemporal expression patterns, using GFP-bound promoters, and how they correlate with the longevity, arrest, and stress resistance phenotypes. Finally, we will determine if rescuing wild type expression levels of these deregulated genes rescues any of the same phenotypes, indicating their importance in that response. Aim 2 will determine the effects of reducing protein synthesis in a tissue-specific manner (hypodermal, intestinal, muscular, or neuronal) using tissue-specific RNAi strains, which will identify where in the worm the longevity pathway is taking place, including revealing how the signaling and cell-cell crosstalk interacts between cell types to confer longevity, arrest, and stress resistance. The second part of Aim 2 involves coalescing information from our spatiotemporal analysis and tissue-specific studies in order to rescue genes found to be important in conferring these survival-promoting phenotypes in order to determine their importance on a cell or tissue-based level. The description of these protein synthesis genes, particularly in relation to their physiological effects, discovered pathways, and correlation with arrest states and stress resistance, will establish a better connection between enhanced longevity and how that longevity is conferred in order to enable the eventual treatment of aging-related ailments and disease.