Functions of the Hypoxia-Induced MicroRNA-210 in Pulmonary Vascular Endothelium

描述（由申请人提供）：一个为期五年的培训计划，建议发展在学术心脏病学的职业生涯，重点是肺血管功能和疾病。主要研究者毕业于医学科学家培训项目，并已完成内科住院医师培训和心脏病学研究员培训（马萨诸塞州总医院，MGH）。Joseph Loscalzo博士将担任主要实验室导师，他是血管生物学领域公认的专家和科学领导者。他成功地培养了许多博士后研究员，其中许多人已经在生物医学科学中担任主要的科学和领导角色。一个由医学专家组成的咨询小组也将提供进一步的科学和职业指导。通过结合多个哈佛附属项目的资源，这种培训环境是理想的培养一个成功的研究计划，在此基础上，一个富有成效的未来职业生涯。 主要研究者已经确定了缺氧诱导的microRNA-210（miR-210）作为缺氧肺动脉内皮细胞线粒体代谢和细胞呼吸的一种新的和必要的调节剂，通过抑制铁硫簇组装蛋白ISCU 1/2。该提议将询问一个模型，其中肺血管系统中内皮特异性表型的控制关键取决于miR-210对ISCU 1/2和铁硫簇的下调。在常氧和缺氧条件下，实验将需要在培养的肺动脉内皮细胞以及鼠受试者的肺脉管系统中表达miR-210和抑制miR-210。表型将通过分子、遗传、生物化学和生物物理技术的组合进行评估。在“特定目的”下列出的拟议实验将阐明miR-210、ISCU 1/2和铁硫簇在以下调节中的作用：1）线粒体电子传递; 2）活性氧通量; 3）一氧化氮生物利用度。结果将提高我们对缺氧肺血管生理和病理生理适应的分子理解，并可能指向新的治疗靶点。 公共卫生相关性：该提案将定义一种新分子（microRNA-210）在调节肺血管细胞对低氧暴露的反应中的关键作用。在这样做的过程中，预计将提高目前的理解的机制，低氧条件影响肺血管在正常和疾病状态，并可能指向未来的治疗目标。

DESCRIPTION (provided by applicant): A five year training program is proposed to develop a career in academic cardiology with a focus on pulmonary vascular function and disease. The principal investigator is a graduate of the Medical Scientist Training Program and has completed residency training in Internal Medicine and fellowship training in Cardiology (Massachusetts General Hospital, MGH). Dr. Joseph Loscalzo will serve as the primary laboratory mentor and is a recognized expert and scientific leader in vascular biology. He has successfully trained numerous postdoctoral fellows, many of whom have gone on to major scientific and leadership roles in biomedical sciences. An advisory panel of expert medical scientists will also provide further scientific and career guidance. By combining the resources of multiple Harvard-affiliated programs, this training environment is ideal to cultivate a successful research program on which to base a productive future career. The principal investigator has identified the hypoxia-induced microRNA-210 (miR-210) as a novel and essential regulator of mitochondrial metabolism and cellular respiration in hypoxic pulmonary arterial endothelial cells, via repression of the iron-sulfur cluster assembly proteins ISCU1/2. This proposal will interrogate a model whereby control of endothelial-specific phenotypes in the pulmonary vasculature depends critically upon the down-regulation of ISCU1/2 and iron-sulfur clusters by miR-210. Under conditions of normoxia and hypoxia, experiments will entail expression of miR-210 and inhibition of miR-210 in cultured pulmonary arterial endothelial cells as well as in the pulmonary vasculature of murine subjects. Phenotypes will be assessed by a combination of molecular, genetic, biochemical, and biophysical techniques. Proposed experiments listed under "Specific Aims" will elucidate the role of miR-210, ISCU1/2, and iron-sulfur clusters in the regulation of: 1) mitochondrial electron transport; 2) reactive oxygen species flux; and 3) nitric oxide bioavailability. Results will improve our molecular understanding of physiologic and pathophysiologic adaptations in the hypoxic pulmonary vasculature and may point to novel therapeutic targets. PUBLIC HEALTH RELEVANCE: This proposal will define the critical actions of a novel molecule (microRNA-210) in regulating the response to low oxygen exposure in cells that line the blood vessels of the lungs. In doing so, it is expected to improve the current understanding of the mechanisms by which low oxygen conditions affect the pulmonary vessels during normal and disease states and may point to future therapeutic targets.