Hypoxic Sensing Transcription Factor EPAS1 in Mice

小鼠缺氧感知转录因子 EPAS1

• 批准号: 6537970
• 负责人: Joseph Anthony Garcia
• 金额: $ 12.26万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-19 至 2005-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6537970
• 关键词: biotechnology; cardiovascular function; environmental adaptation; gene targeting; genetically modified animals; hypoxia; laboratory mouse; technology /technique development; transcription factor; vascular endothelium

DESCRIPTION (provided by applicant) The overall goal of this proposal is to increase our understanding of the hypoxic response in the adult mouse. Members of the PAS domain family of transcription factors have important roles in development and in response to environmental stresses including hypoxic-responsiveness. The first identified hypoxic-responsive member of this family, hypoxia inducible factor I (HIFI), activates a number of target genes including the erythropoeitin promoter. Endothelial PAS domain protein 1 (EPAS1), the second hypoxic-responsive member of the PAS family, is expressed at high levels in vascular endothelial cells, carotid body glomus cells, and pulmonary pneumocytes. Despite the observation that HIF1 and EPAS1 can bind to similar DNA regulatory sequences in model promoters, it is likely that HIF1 and EPAS1 play distinctive regulatory roles as evident by the differences in sites of expression as well as stage and description of embryonic lethality for the respective knockouts. We hypothesize that hypoxic regulators such as EPAS1 are fundamentally involved in the pathogenesis underlying hypertension, sleep apnea, and heart failure. The specific aims of this project are to: (1) construct a preconditional strain for use in generating conditional knockouts for EPAS1; (2) construct transgenic mice that express cre in a tissue-restricted manner in anatomic sites that overlap EPAS1 expression; (3) generate knockout mice lacking EPAS1 in endothelial or glomus cells and characterize the cardiovascular function of the resultant EPAS1- deficient mice under normoxic as well as intermittent hypoxic conditions. The principal investigator has completed a residency in internal medicine, a fellowship in cardiology, and most recently a postdoctoral fellowship in the department of biochemistry. His doctoral research involved molecular biological studies of HIV gene regulation. His postdoctoral research involved defining the neurobiological role of a neural-restricted transcription factor in a mouse knockout model. He has just recently been appointed an assistant professor in the cardiology division. The proposed research on physiologic aspects of cardiovascular function will result in the acquisition of research skills in experimental areas novel to this investigator. His sponsors and advisory committee members will serve as important resources due to their expertise in cardiovascular biology and in other areas directly relevant to the pursuit of these experimental aims. The CIDA grant would aid in development of the principal investigator into an independent research leader in cardiovascular biology whose career goals are to study signal transduction in the adult mammalian cardiovascular system.

描述（由申请人提供） 本提案的总体目标是增进我们对 成年小鼠的缺氧反应。 PAS结构域家族的成员 转录因子在发育和应答 环境压力，包括缺氧反应。第一识别 该家族缺氧反应性成员，缺氧诱导因子I（HIFI）， 激活许多靶基因，包括促红细胞生成素启动子。 内皮PAS结构域蛋白1（EPAS 1），第二个缺氧反应成员 PAS家族成员，在血管内皮细胞中高水平表达， 颈动脉体血管球细胞和肺细胞。 尽管观察到 HIF 1和EPAS 1可以与模型中相似的DNA调控序列结合， 启动子，可能是HIF 1和EPAS 1发挥独特的调控作用， 正如表达部位和阶段的差异所证明的那样， 对各个敲除的胚胎致死率的描述。 我们 假设低氧调节因子如EPAS 1在根本上参与了 高血压、睡眠呼吸暂停和心力衰竭的发病机制。 本项目的具体目标是：（1）构建一个预条件 用于产生EPAS 1条件性敲除菌株;（2）构建体 在解剖学上以组织限制性方式表达cre的转基因小鼠 与EPAS 1表达重叠的位点;（3）产生缺乏EPAS 1的敲除小鼠 在内皮细胞或血管球细胞中，并表征 将所得EPAS 1缺陷小鼠在常氧以及间歇性 低氧条件下。 主要研究者已完成内科住院医师培训， 在心脏病学奖学金，最近在博士后奖学金， 生物化学系。 他的博士研究涉及分子 HIV基因调控的生物学研究。 他的博士后研究涉及 定义神经限制性转录因子的神经生物学作用 在小鼠敲除模型中。 他最近刚被任命为助理 心脏科的教授 生理学研究的建议 心血管功能方面将导致获得研究 新的实验领域的技能。 他的赞助商和 咨询委员会成员将作为重要资源，因为他们 心血管生物学和其他直接相关领域的专业知识， 这些实验目标的实现。 加拿大国际开发署的赠款将有助于 将主要研究者培养为独立的研究领导者 他的职业目标是研究信号传导 在成年哺乳动物的心血管系统中。