Erythrocyte Nitrosothiol Flux and Vasoregulation in Lung

红细胞亚硝基硫醇通量和肺血管调节

• 批准号: 7009990
• 负责人: ALLAN DOCTOR
• 金额: $ 6.72万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-01 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7009990
• 关键词: SDS polyacrylamide gel electrophoresis; biological signal transduction; cell membrane; clinical research; erythrocytes; hemodynamics; hemoglobin; human subject; immunocytochemistry; inflammation; lung; nitric oxide; nitroso compounds; proteomics; pulmonary circulation; respiratory hypoxia; respiratory insufficiency /failure; respiratory pharmacology; tissue /cell culture; vascular endothelium

DESCRIPTION (provided by applicant): This proposal describes a five-year plan for the attainment of research skills necessary in the career development of an academic physician-scientist in Critical Care. The candidate is a junior faculty member in an established Division of Pediatric Critical Care and has preliminary research training in pulmonary physiology. The award will be utilized to broaden this background by acquiring a foundation in vascular cell biology and the biochemistry of nitrosative signaling to enable independent investigation of erythrocytic and endothelial communication in the pathobiology of disrupted vasomotor control. Ben Gaston, MD, a leader in pulmonary nitrosothiol signaling research, will serve as primary mentor for the award and Brian Duling, PhD, a pioneer in microcirculation research and vascular cell biology will serve as Co-Mentor. Specific expertise in nitrosative hemoglobin chemistry will be provided by collaboration with investigators from Duke and the University of Pennsylvania. Additionally, an advisory committee of senior scientists will provide scientific and career guidance. Research conducted with this award will investigate the role of the erythrocyte as a link between dysregulated pulmonary blood flow and remote inflammation via nitric oxide (NO) and hemoglobin (Hb) interactions. There is evidence for a nitrosative signaling network in which Hb and NO reactions are balanced to transduce regional redox gradients, coupling oxygen tension and the distribution of NO (and thus flow), in vascular beds. In this regard, we hypothesize oxidative stress in the systemic inflammatory response syndrome (SIRS) may disrupt normal erythrocytic nitrosative signaling and explain dysregulated pulmonary blood flow in this state. We aim, in this project, to determine (1) the degree of abnormal NO loading of RBCs in SIRS, correlating with onset and severity of respiratory failure and (2) to determine the change in Hb vasoactivity in the lung following addition of NO to a beta-chain cysteine (betacys93), the allosteric control of this change, and (3) the endothelial regulation of Hb-based nitrosative signaling. We will pursue these goals on three levels: (1) molecular investigation of intraerythrocytic Hb approximately NO chemistry (2) pharmacologic, immunohistochemical, and proteomic investigations of NO signaling between erythrocytes and endothelial cells in culture, and (3) physiologic correlation of our findings in the isolated murine lung, permitting further transgenic and pharmacologic query. At the conclusion of this project, we expect to define the mechanism of NO traffic between remote vascular beds and the lung via the erythrocyte, as effected by serial transnitrosation reactions. At the conclusion of the development period, the candidate will have acquired skills to pursue further independent investigation of (S)NO and erythrocyte vasoactivity with the hope of informing therapy directed at the dysregulation of regional blood flow in the lung. The candidate's long-term goal is to define the mechanism of NO traffic between regional vascular beds via the erythrocyte, and its role in the evolution of multiple organ failure in severe inflammatory states.

描述（由申请人提供）：该提案描述了一个五年计划，以实现在重症监护学术医师和科学家的职业发展中所必需的研究技能。该候选人是一名已成立的儿科重症监护室的初级教员，并在肺生理学方面接受过初步的研究培训。该奖项将通过获得血管细胞生物学和亚硝化信号生物化学的基础来扩大这一背景，以便在血管运动控制中断的病理生物学中对红细胞和内皮细胞通讯进行独立研究。肺亚硝基硫醇信号研究的领导者Ben Gaston医学博士将担任该奖项的主要导师，微循环研究和血管细胞生物学的先驱Brian Duling博士将担任联合导师。与杜克大学和宾夕法尼亚大学的研究人员合作，将提供亚硝化血红蛋白化学方面的专业知识。此外，一个由资深科学家组成的咨询委员会将提供科学和职业指导。该奖项的研究将通过一氧化氮（NO）和血红蛋白（Hb）的相互作用，研究红细胞在肺血流失调和远端炎症之间的作用。有证据表明，在一个亚硝化信号网络中，Hb和NO反应被平衡以传导区域氧化还原梯度，耦合氧张力和NO的分布（因此流动），在血管床中。在这方面，我们假设系统性炎症反应综合征（SIRS）中的氧化应激可能会破坏正常的红细胞亚硝酸盐信号，并解释这种状态下肺血流失调的原因。在这个项目中，我们的目标是确定(1)SIRS中红细胞异常NO负荷的程度，与呼吸衰竭的发病和严重程度相关；(2)确定在β链半胱氨酸（betacys93）中添加NO后肺Hb血管活性的变化，这种变化的变张控制；(3)基于Hb的亚硝化信号的内皮调节。我们将在三个层面上追求这些目标：(1)红细胞内Hb近似NO化学的分子研究；(2)培养红细胞和内皮细胞之间NO信号的药理学、免疫组织化学和蛋白质组学研究；(3)分离小鼠肺中我们的发现的生理学相关性，允许进一步的转基因和药理学研究。在本项目结束时，我们期望确定通过红细胞在远程血管床和肺之间的NO运输机制，受一系列亚硝化反应的影响。在开发阶段结束时，候选人将获得进一步独立研究(S)NO和红细胞血管活性的技能，希望为针对肺局部血流失调的治疗提供信息。该候选人的长期目标是确定通过红细胞在区域血管床之间运输NO的机制，及其在严重炎症状态下多器官衰竭演变中的作用。