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: 6839438
• 负责人: ALLAN DOCTOR
• 金额: $ 12.14万
• 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 2008-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6839438
• 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）：该提案描述了一项为期五年的计划，以实现重症监护学术医师科学家职业发展所必需的研究技能。该候选人是小儿重症监护阶层既定部门的初级教师，并接受了肺部生理学的初步研究培训。该奖项将通过获得血管细胞生物学的基础和硝化信号传导的生物化学来扩大这种背景，以在干扰血管瘤控制的病理生物学中独立研究红细胞和内皮交流。医学博士本·加斯顿（Ben Gaston）是肺硝基硫醇信号研究的领导者，将担任该奖项的主要导师，而Brian Duling，博士学位，微循环研究的先驱和血管细胞生物学的先驱将担任同事。与杜克大学和宾夕法尼亚大学的调查人员合作将提供硝化血红蛋白化学方面的具体专业知识。此外，高级科学家的咨询委员会将提供科学和职业指导。通过该奖项进行的研究将研究红细胞通过一氧化氮（NO）和血红蛋白（HB）相互作用的肺血流和远程炎症之间的联系。有证据表明，硝化信号网络，其中Hb和无反应在局部氧化还原梯度，耦合氧张力以及NO（因此流动）的分布中，在血管床中平衡。在这方面，我们假设系统性炎症反应综合征（SIR）中的氧化应激可能破坏正常的红细胞硝化信号传导，并解释该状态下肺血流失调。在这个项目中，我们的目标是确定（1）SIRS中RBC的异常NO载荷程度，与呼吸道衰竭的发作和严重程度相关，以及（2）确定在肺链囊cySteine（Betacys93）中添加NO后肺中HB血管活性的变化，该型（Betacys93），noterial noterive no no no no no no no（Betacys93），noterial no no no no no no no no no no no no no n n n n n n n n n n n n no no no no no no no。信号。 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.在该项目的结论结束时，我们希望通过串行的转硝化反应的影响来定义远程血管床与肺之间无流量的机制。在开发期结束时，候选人将获得对NO和红细胞血管活性的进一步独立研究，以期为肺部区域血流失调的治疗提供信息。候选人的长期目标是通过红细胞来定义区域血管床之间没有交通的机制，及其在严重炎症状态下多器官衰竭演变中的作用。