Conserved longevity mechanisms of the hypoxic response pathway

描述（申请人提供）：本研究项目主要研究应激反应途径及其在衰老过程中的作用。更具体地说，它集中于缺氧诱导因子-1 (HIF-1)，这是一种高度保守的转录因子，参与对低氧的反应。最近的报道将HIF-1置于秀丽隐杆线虫中新发现的年龄相关通路的核心。有趣的是，敲除HIF-1抑制因子vhl-1可以显著延长蠕虫的寿命，但却会导致人类患上一种疾病（von Hippel Lindau病）。我的研究重点是HIF-1的稳定如何对蠕虫的寿命产生积极影响，研究与寿命影响有关的组织和下游因素，并将它们与导致人类疾病的因素区分开来。由于像HIF-1这样的应激反应途径是高度保守的，因此很可能至少有一些蠕虫中的下游效应物会出现在哺乳动物中。在表征这些下游基因时，我将使用生化技术，包括一种新的方法来测量HIF-1的共价变化。这种方法的发展将回答关于HIF-1是如何被调节的问题，并为在蠕虫和其他生物中进行HIF-1和其他应激反应蛋白的类似研究提供蓝图。在上述实验的结论中，我将证明HIF-1在一组已知细胞中的稳定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。在此过程中，我将开发一种方案来测量HIF-1对已知转录抑制因子的翻译后修饰，并计划在未来的研究中扩展这些实验。因此，这些研究对于确定应激反应途径在长寿中的作用将是重要的。这个过渡性奖项将包括我博士后研究的最后两年和我独立的教师培训的前三年。在提案的指导阶段，我将定义组织并学习表征hif -1介导的寿命所需的下游蛋白质的技术，包括开发测量其翻译后修饰的方法。在过渡到独立后，我将更全面地表征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，该提案也关注职业发展的许多方面。提案的指导部分和独立部分都强调在课堂和实验室进行培训，同时继续撰写和审查论文和拨款申请。总之，这个项目将有助于确定缺氧反应途径如何改善哺乳动物的健康寿命和寿命，同时也提供了在指导阶段进行训练的机会，将我的工作与导师的工作区分开来，并制定新的实验方案。

DESCRIPTION (provided by applicant): 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 thse 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 th 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.