Developing EPR tools for preclinical interrogation of redox regulation mechanisms contributing to acute lung injury

开发 EPR 工具，用于临床前询问导致急性肺损伤的氧化还原调节机制

基本信息

批准号：
10396627
负责人：
SANDRA S. EATON
金额：
$ 37.7万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-22 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10396627
关键词：
Acute Lung Injury Acute Respiratory Distress Syndrome Address Adult Alveolar Antioxidants Automobile Driving Autopsy Biology Biopsy Bleomycin Blood Circulation Breathing COVID-19 pandemic Cells Cellular Stress Child Clinical Clinical Research Critical Illness DNA Data Development Disease Disulfides Electron Spin Resonance Spectroscopy Electrons Environment Fibrosis Fluorescent Probes Free Radicals Future Generations Genes Glutathione Glutathione Disulfide Goals Gold Homeostasis Human Hydroxylamine Image Imaging technology Incidence Individual Inflammation Life Link Lipids Lung Lung diseases Magnetic Resonance Imaging Measurement Measures Methodology Methods Mitochondria Molecular Monitor Morbidity - disease rate Mouse Strains Mus Nitrogen Noise Operative Surgical Procedures Organ Organ Transplantation Oxidants Oxidation-Reduction Oxidative Stress Oxygen Pathogenesis Pathway interactions Periodicity Pre-Clinical Model Process Production Proteins Pulmonary Fibrosis Radio Waves Regulation Reporting Research Route Sampling Scanning Signal Transduction Signaling Molecule Site Spin Trapping Structure of parenchyma of lung Sulfhydryl Compounds Superoxide Dismutase Superoxides Testing Therapeutic Time Tissues Variant Visualization antioxidant enzyme antioxidant therapy autooxidation base effective therapy end stage disease endophenotype ex vivo imaging extracellular human disease imager improved improved outcome in vivo insight knock-down mortality mouse model new technology novel organ injury overexpression oxidation pre-clinical pulmonary vascular disorder response to injury spectroscopic imaging tool treatment response

项目摘要

PROJECT SUMMARY Acute respiratory distress syndrome (ARDS), the most severe form of acute lung injury (ALI), is a principal cause of life-threatening illness in both adults and children. Unfortunately, despite decades of research in search of effective treatments, the high mortality of ARDS has remained largely unchanged, requiring breakthroughs in the methodologies to assess individuals with ARDS to guide therapeutic strategies. Oxidative stress is critical in the pathogenesis of ARDS (as well as numerous other human diseases); however, the clinical utility of antioxidant therapies is complicated because reactive oxygen and nitrogen species produced at low and controlled levels are also signaling molecules essential for cellular homeostasis and adaptation to cellular stress through targeted specific redox-sensitive pathways. Based on these findings, “oxidative stress” has been redefined from a simple imbalance in oxidants and antioxidants to also include a disruption in redox signaling and control. This revised definition is driving improved tools and therapeutic approaches in the field of Redox Biology; these advances provide new opportunities to understand and treat the disruption of redox-regulated processes that contribute to pathogenesis of ARDS. One major barrier to the study of dysregulated redox signaling in human ARDS is the lack of rigorous methodologies to precisely determine the redox status of the lung in vivo. This proposal addresses this major gap by developing new technology to measure the redox status of the acutely injured lung in vivo using electron resonance molecular that react EPR probes We hypothesize that advanced applications of Electron Paramagnetic Resonance (EPR) spectroscopy to preclinical models using rapid scan in vivo EPR imaging will provide novel insight into the time-course and localization of free radical production and redox status in lung disease. We will use EPR spin probes that can differentiate between total cell and mitochondrial superoxide as well as a novel spin probe that can detect the redox status of intracellular glutathione, providing precise and specialized measures of the redox status in mice with different levels of oxidative stress. We paramagnetic (EPR) imaging spectroscopy is the gold-standard for the measurement of free radicals: probes known as “spin traps” react with specific short-lived free radicals to form more stable radicals can then be easily detected and quantified by the EPR spectrometer, while “spin probe” molecules that with thiol species such as glutathione can report cellular thiol redox status. Importantly, like MRI, the imaging spectrometer uses radio waves that readily penetrate tissue and enable visualization of the spin within the mouse organs. . EPR boldly propose assignment essential, of endophenotypes in ARDS and inform future clinical studies and therapeutic decisions. that this will provide previously unattainable information that will guide the

项目概要急性呼吸窘迫综合征 (ARDS) 是急性肺损伤 (ALI) 的最严重形式，不幸的是，尽管几十年来，这是导致成人和儿童危及生命的疾病的主要原因。寻找有效治疗方法的研究，ARDS的高死亡率基本保持不变，需要在评估 ARDS 个体的方法上取得突破，以指导治疗策略。氧化应激在 ARDS（以及许多其他人类疾病）的发病机制中至关重要；然而，抗氧化疗法的临床应用很复杂，因为活性氧和氮以低水平和受控水平产生的物种也是细胞稳态所必需的信号分子基于这些发现，通过有针对性的特定氧化还原敏感途径来适应细胞应激。 “氧化应激”已从简单的氧化剂和抗氧化剂不平衡重新定义为还包括这一修订后的定义正在推动工具和治疗方法的改进。氧化还原生物学领域的方法；这些进展为理解和治疗提供了新的机会氧化还原调节过程的破坏导致 ARDS 的发病机制之一。对人类 ARDS 中氧化还原信号失调的研究缺乏精确的严格方法该提案通过开发新的技术来解决这一主要差距。利用电子测量体内急性损伤肺氧化还原状态的技术谐振分子那反应 EPR 探针我们采取了电子顺磁的先进应用使用快速扫描体内 EPR 成像对临床前模型进行共振 (EPR) 光谱分析为自由基产生和氧化还原状态的时间进程和定位提供新颖的见解在肺部疾病中，我们将使用可以区分总细胞和线粒体的 EPR 自旋探针。超氧化物以及一种新型自旋探针可以检测细胞内谷胱甘肽的氧化还原状态，提供对具有不同氧化应激水平的小鼠的氧化还原状态进行精确和专门的测量。顺磁的 (EPR) 成像光谱是自由基测量的黄金标准：被称为“自旋陷阱”的探针与特定的短命自由基发生反应，形成更稳定的自由基然后可以通过 EPR 光谱仪轻松检测和定量，而“自旋探针”分子重要的是，与谷胱甘肽等硫醇种类可以报告细胞硫醇氧化还原状态，就像 MRI 一样。成像光谱仪使用易于穿透组织并实现旋转可视化的无线电波小鼠器官内。 .EPR 大胆提议任务基本的， ARDS 的内表型并为未来的临床研究和治疗决策提供信息。这将提供以前无法获得的信息来指导