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

描述(由申请者提供)：一个为期五年的培训计划是为了发展一个学术心脏病学的职业生涯，重点是肺血管功能和疾病。首席研究员毕业于医学科学家培训计划，并已完成内科住院医师培训和心脏病学研究员培训(麻省总医院，MGH)。约瑟夫·洛斯卡尔佐博士将担任主要实验室导师，他是公认的血管生物学专家和科学领导者。他成功地培养了许多博士后研究员，其中许多人后来在生物医学科学方面担任了重要的科学和领导职务。一个由专家医学家组成的顾问小组也将提供进一步的科学和职业指导。通过整合多个哈佛附属项目的资源，这种培训环境是培养一个成功的研究项目的理想选择，为未来富有成效的职业生涯奠定了基础。主要研究人员发现，低氧诱导的microRNA-210(miR-210)通过抑制铁-硫簇组装蛋白ISCU1/2，是低氧肺动脉内皮细胞线粒体代谢和细胞呼吸的新的和必要的调节因子。这项提议将询问一个模型，在该模型中，肺血管内皮细胞特异性表型的控制在很大程度上取决于miR-210下调ISCU1/2和铁-硫簇的表达。在常氧和低氧条件下，实验将需要在培养的肺动脉内皮细胞和小鼠肺血管中表达miR-210并抑制miR-210。表型将通过分子、遗传、生化和生物物理技术的组合进行评估。在“特定目标”下列出的拟议实验将阐明miR-210、ISCU1/2和铁-硫簇在调节以下方面的作用：1)线粒体电子传递；2)活性氧物种通量；以及3)一氧化氮的生物利用度。结果将提高我们对缺氧性肺血管的生理学和病理生理学适应的分子理解，并可能指向新的治疗靶点。 与公共健康相关：这项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。通过这样做，预计将改善目前对低氧条件在正常和疾病状态下影响肺血管的机制的理解，并可能指明未来的治疗目标。

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.