Genetic pathway and cellular mechanism underlying organismic responses to hypoxia and hypothermia

项目概要/摘要 适当的温度和氧气水平可以实现基本的生命活动。低温（低温）和 氧气水平降低（缺氧）普遍影响基本的生化过程、细胞 新陈代谢、有机体生理学和行为。缺氧和氧化应激也是重要特征 缺血性疾病，包括中风和心脏病发作，其治疗可以极大地受益于新兴的 “低温治疗”的程序。我们的实验室对基础遗传分析和 弹性生物体内缺氧、低温、先天缺血耐受性的机制研究，以及 细胞保护，防止代谢应激引起的组织损伤。我们使用 1) 遗传上易于驯化的线虫 从大规模筛选中分离出的具有异常细胞生理和有机行为的突变体 缺氧/低温反应的表型和 2) 红树林鳉鱼，唯一已知的自体受精 具有与秀丽隐杆线虫相似的遗传学和已知的与线虫相关的极端生理表型的脊椎动物 缺氧和低温作为发现工具。此外，我们还离体培养哺乳动物神经干细胞 从冬眠的地松鼠中分离出来，以揭示缺氧/低温的细胞内在机制 宽容。通过多学科方法和技术，我们一直在进行富有成效的研究 计划并已经发现了基因、蛋白质变体和途径的新作用机制 赋予细胞保护和对缺氧和低温的机体反应。在此 R35 应用程序中，我们 建议继续进行这些易于处理和创新的调查，以扩大我们对如何进行的基本了解 细胞和生物体应对缺氧和低温，以表征新的基因和途径 从我们的正向遗传和 RNAi 筛选中鉴定出，并确定先天性的关键遗传决定因素 弹性生物体的缺氧/缺血耐受性。 PI和实验室丰富的先前经验和 多样化但互补的模型系统方面的专业知识非常适合执行并成功 在旧金山加州大学心血管研究所完成项目 （加州大学旧金山分校）。由于 MIRA R35 旨在“整合 NIGMS 对多个项目的支持，这些项目可能 就像我们的情况一样，我们将平衡专用于每个模型系统的努力和资源， 它们同样容易解决我们研究计划中的相同核心问题。

Project Summary/Abstract Proper temperature and oxygen levels enable essential life activities. Low temperature (hypothermia) and reduced level of oxygen (hypoxia) pervasively influence fundamental biochemical processes, cellular metabolism, organismic physiology and behaviors. Hypoxia and oxidative stresses are also key features in ischemic disorders, including stroke and heart attack, treatment of which can greatly benefit from the emerging procedure of “therapeutic hypothermia.” Our laboratory is interested in fundamental genetic analysis and mechanistic studies of hypoxia, hypothermia, innate ischemic tolerance in resilient organisms, and cytoprotection against tissue injuries caused by metabolic stresses. We use 1) genetically tractable C. elegans mutants isolated from large-scale screens with abnormal cell physiological and organismic behavioral phenotypes in hypoxia/hypothermia responses and 2) Mangrove Killifish, the only known self-fertilizing vertebrate with genetics similar to that of C. elegans and known extreme physiological phenotypes related to hypoxia and hypothermia, as discovery tools. In addition, we culture mammalian neural stem cells ex vivo isolated from hibernating ground squirrels to unravel cellular intrinsic mechanisms of hypoxia/hypothermia tolerance. With multidisciplinary approaches and technologies, we have been running a productive research program and already discovered novel mechanisms of action of genes, protein variants and pathways in conferring cytoprotection and organismic responses to hypoxia and hypothermia. In this R35 application, we propose to continue these tractable and innovative lines of inquiries to expand our basic understanding of how cells and organisms cope with hypoxia and hypothermia, to characterize novel genes and pathways already identified from our forward genetic and RNAi screens, and to identify key genetic determinants of innate hypoxia/ischemic tolerance in resilient organisms. The PI and laboratory's extensive prior experience and expertise in diverse but complementary model systems are well suited for executing and successfully completing the project in the Cardiovascular Research Institute at the University of California, San Francisco (UCSF). As the MIRA R35 is intended to “enable consolidation of NIGMS support for multiple projects that may be disparate” as is our case, we will balance efforts and resources dedicated to each of the model systems, which are similarly tractable towards addressing the same core questions in our research program.