Conserved longevity mechanisms of the hypoxic response pathway

该研究项目的重点是应激反应途径及其在衰老过程中的作用。更 特别是，它集中在低氧诱导因子-1（HIF-1），一个高度保守的转录因子 与低氧有关。最近的报告将HIF-1置于新发现的与年龄相关的疾病的核心 在线虫线虫Caelophabditis elegans途径。有趣的是，敲除HIF-1阻遏物vhl-1 显著增加蠕虫的寿命，但在人类中引起疾病（von Hippel Lindau病）。我 研究的重点是HIF-1的稳定性如何积极影响蠕虫的寿命，研究相关的 组织和下游因素所必需的寿命的影响，并将它们从那些导致 人的疾病。由于像HIF-1这样的应激反应途径是高度保守的，很可能至少 蠕虫中的一些下游效应物将存在于哺乳动物中。在描述这些下游的 基因，我将使用生物化学技术，包括一种新的方法来衡量共价变化， HIF-1。这种方法的发展将回答有关HIF-1是如何调节的问题，并提供一种新的方法。 这为在蠕虫和其他生物中的HIF-1和其他应激反应蛋白的类似研究提供了蓝图。在 作为所描述的实验的结论，我将证明HIF-1在一组已知的细胞中的稳定性可以 通过激活特定细胞中的特定蛋白质来增加蠕虫的健康寿命和寿命。我将描述 这些蛋白质的功能以及它们是否位于任何其他衰老干预措施的下游。为了做到这一点，我将 开发一种方案来测量HIF-1翻译后修饰，以响应已知的转录 阻遏物，并计划在未来的研究中扩大这些实验。因此，这些研究将对 定义压力反应途径在长寿中的作用。这一过渡性奖项将包括最后两个 我的博士后研究和前三年的独立，教师培训。期间 在建议的指导阶段，我将定义组织，并学习技术来表征 HIF-1介导的长寿所必需的下游蛋白，包括开发用于 测量其翻译后修饰。在过渡到独立后，我将更全面地描述 HIF-1信号通路及其下游蛋白，重点是最终将工作过渡到 哺乳动物模型。作为一个过渡性奖项，这个建议也关注了职业生涯的许多方面 发展建议中的辅导部分和独立部分都强调在以下方面的培训： 在课堂和实验室，同时继续撰写和审查论文和资助申请。在 总之，该项目将有助于确定缺氧反应途径如何改善健康状况， 哺乳动物的寿命，同时也提供了在指导阶段，分离和 将我的工作与我导师的工作区分开来，并开发出一种新的实验方案。

This research project focuses on stress-response pathways and their role in the aging process. More specifically, it concentrates on the hypoxia-inducible factor-1, (HIF-1), a highly conserved transcription factor involved in response to low oxygen. Recent reports have placed HIF-1 at the core of a newly discovered age-related pathway in the nematode Caenorhabditis elegans. Interestingly, knocking out the HIF-1 repressor vhl-1 significantly increases lifespan in worms, but causes a disease (von Hippel Lindau disease) in humans. My research focuses on how HIF-1 stabilization positively affects longevity in worms, studying the associated tissues and downstream factors necessary for lifespan effects and separating them from those that cause disease in people. Since stress response pathways like HIF-1 are highly conserved, it is likely that at least some of the downstream effectors in worms will be present in mammals. In characterizing these downstream genes, I will use biochemical techniques including a novel approach to measure covalent changes made to HIF-1. The development of this approach will answer questions about how HIF-1 is regulated and provide a blueprint for similar studies in HIF-1 and other stress-response proteins in worms and other organisms. At the conclusion of the described experiments, I will show that HIF-1 stabilization in a known set of cells can increase worm healthspan and longevity by activating specific proteins in specific cells. I will characterize the function of these proteins and whether they lie downstream of any other aging interventions. In doing this, I will develop a protocol to measure HIF-1 post-translational modifications in response to a known transcriptional repressor, and plan to expand these experiments in future studies. Thus, these studies will be important for defining the role of stress response pathways in longevity. This transitional award will encompass the final two years of my postdoctoral research and the first three years of my independent, faculty training. During the mentored stage of the proposal, I will define the tissues and learn the techniques to characterize the downstream proteins necessary for HIF-1-mediated longevity, including developing the methodology for measuring its post-translational modifications. After transitioning to independence, I will more fully characterize the HIF-1 signaling pathway and its downstream proteins with a focus on eventually transitioning the work into mammalian models. As a transitional award, this proposal also focuses on many aspects of career development. Both the mentored and independent portions of the proposal emphasize training in the classroom and in the laboratory while continuing to write and review papers and grant applications. In summary, this project will help define how the hypoxic response pathway could improve healthspan and lifespan in mammals while also providing opportunities for training during the mentored stage, separating and distinguishing my work from that of my mentor, and developing a new experimental protocol.