Hypoxic Sensing Transcription Factor EPAS1 in Mice

描述（由申请人提供） 该提案的总体目标是增加我们对 成年小鼠的缺氧反应。 PAS 域家族的成员 转录因子在发育和响应中具有重要作用 环境压力，包括缺氧反应。第一个确定的 该家族的缺氧反应成员，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 (EPAS1)，第二个缺氧反应成员 PAS 家族的成员，在血管内皮细胞中高水平表达， 颈动脉体血管球细胞和肺泡细胞。 尽管观察 HIF1 和 EPAS1 可以与模型中相似的 DNA 调控序列结合 启动子，HIF1 和 EPAS1 可能发挥独特的调节作用 表达位点以及阶段和阶段的差异显而易见 各个敲除的胚胎致死率的描述。 我们 假设 EPAS1 等缺氧调节因子从根本上参与其中 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）构建一个前提条件 用于产生 EPAS1 条件性敲除的菌株； (2)构建 在解剖学中以组织限制方式表达 cre 的转基因小鼠 与 EPAS1 表达重叠的位点； (3) 生成缺乏EPAS1的敲除小鼠 在内皮细胞或血管球细胞中并表征心血管功能 由此产生的 EPAS1 缺陷小鼠在常氧和间歇状态下 缺氧条件。 主要研究者完成了内科住院医师实习， 心脏病学研究员，最近是博士后研究员 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 定义神经限制转录因子的神经生物学作用 在小鼠敲除模型中。 他最近刚刚被任命为助理 心脏病学系教授。 拟议的生理学研究 心血管功能方面将导致获得研究成果 实验领域的技能对于该研究者来说是新颖的。 他的赞助商和 咨询委员会成员将作为重要资源，因为他们的 心血管生物学和其他直接相关领域的专业知识 追求这些实验目标。 CIDA 赠款将有助于 首席研究员发展成为独立研究领导者 心血管生物学专业，其职业目标是研究信号转导 在成年哺乳动物的心血管系统中。

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.