New Oxidation-Sensing Probes to Evaluate Mitochondrial Dysfunction in Lung Injury

用于评估肺损伤中线粒体功能障碍的新型氧化传感探针

基本信息

批准号：
9513774
负责人：
Cristina Maria Furdui
金额：
$ 51.63万
依托单位：
WAKE FOREST UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-07-14 至 2020-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9513774
关键词：
ATP Synthesis Pathway Acetyl Coenzyme A Aging Animal Model Animals Antibodies Biological Biological Markers Biology Biotin Breathing Cell Communication Cell Culture Techniques Cell Nucleus Cell model Cells Chemicals Citric Acid Cycle Communication Communities Computer Simulation Computing Methodologies Cysteine DNA Damage Data Data Set Dependence Detection Development Disease Down-Regulation Energy Metabolism Environmental Exposure Environmental Risk Factor Epigenetic Process Epithelial Cells Event Exposure to Financial Support Human Human body Image In Situ In Vitro Inflammation Investigation Ionizing radiation Kinetics Label Ligation Link Location Malignant Neoplasms Mass Spectrum Analysis Measurement Medicine Membrane Potentials Metabolic Methods Mitochondria Mitochondrial Proteins Modification Molecular Molecular Target Monitor Movement Mus Mutagenesis Organelles Oxidation-Reduction Oxidative Stress Oxides Pathogenesis Pathway interactions Peptides Phase Phenotype Phosphines Population Process Property Proteins Proteomics Publications Radiation therapy Reactive Oxygen Species Reagent Recombinant Proteins Reporting Research Research Design Respiration Respiratory physiology Series Signal Transduction Silver Site Stress Study models Sulfenic Acids Technology Testing United States National Institutes of Health Validation Western Blotting base chromatin remodeling data integration detection of nutrient environmental stressor forest imaging modality interest lung injury mitochondrial dysfunction mitochondrial membrane nanoparticle oxidation programs protein E public health relevance response single molecule tool trafficking uptake

项目摘要

DESCRIPTION (provided by applicant): Mitochondrial dysfunction and accumulation of reactive oxygen species (ROS) in environmentally associated diseases is well established. Yet we have only a limited appreciation for the molecular mechanisms linking these processes to changes in cellular phenotype underlining the pathogenesis of environmental stressors. With previous support from the NIH IMAT program, this research team at Wake Forest has pioneered the development of highly specific chemical probes, which enable detection and identification of oxidized proteins (molecular targets of ROS). While these probes have been used successfully to identify global targets of oxidation within cellular proteins under numerous disease conditions (e.g., cancer, aging, inflammation), they have not yet been targeted to specific organelles within the cells or applied to study cellular response to environmental stressors. The current proposal describes new strategies to achieve these important tasks by focusing first (R21 phase) on the development and validation of mitochondria-targeted chemical probes for protein oxidation and then (R33 phase) on the application of these oxidation-sensing probes and methods of analysis to investigate mechanisms of lung Injury induced by ionizing radiation(IR) and silver nanoparticles (AgNP). The new probes will enable selective labeling of electrophilic and nucleophilic protein sulfenic acids (-SOH) in mitochondria. New imaging methods that combine the mitochondria-targeted and oxidation-sensing probes with an antibody against the protein of interest will be employed to visualize selective protein -SOH modification in situ and movement of the oxidized protein within the cell (e.g., between mitochondria and nucleus). The probes will then be employed mechanistically to investigate the relationship between mitochondrial dysfunction and environmental lung injury. New computational methods (COSMro) will be employed to infer mitochondria-dependent up or downregulation of specific pathways, which will then be validated using studies in cells and animal models of lung injury. These studies will be performed in young and old animals using single and combined environmental stressors to mimic to the extent possible the environmental exposure in a human population. Successful completion of this project will have high impact, enabling a much deeper understanding of mitochondria- and redox-controlled intracellular processes involved in the biological response to environmental stressors encountered in our daily lives.

描述（由申请人提供）：环境相关疾病中的线粒体功能障碍和活性氧（ROS）积累已得到充分证实，但我们对将这些过程与强调环境发病机制的细胞表型变化联系起来的分子机制了解有限。在 NIH IMAT 项目的先前支持下，维克森林大学的这个研究团队率先开发了高度特异性的化学探针，该探针能够检测和识别氧化蛋白（ROS 的分子靶标）。这些探针已成功用于识别多种疾病条件（例如癌症、衰老、炎症）下细胞蛋白质内氧化的全局目标，但它们尚未靶向细胞内的特定细胞器或应用于研究细胞对环境应激源的反应当前的提案描述了实现这些重要任务的新策略，首先（R21阶段）关注用于蛋白质氧化的线粒体靶向化学探针的开发和验证，然后（R33阶段）关注这些技术的应用氧化传感探针和分析方法，用于研究电离辐射（IR）和银纳米粒子（AgNP）引起的肺损伤的机制。新探针将能够选择性标记线粒体中的亲电和亲核蛋白亚磺酸（-SOH）。将线粒体靶向和氧化传感探针与针对感兴趣蛋白质的抗体相结合的成像方法将用于可视化选择性蛋白质-SOH修饰然后，将机械地使用探针来研究线粒体功能障碍和环境性肺损伤之间的关系。 -特定途径的依赖性上调或下调，然后通过肺损伤细胞和动物模型的研究来验证这些研究将在年轻和年老的动物中使用单一和组合的环境压力源来尽可能模拟环境暴露。在该项目的成功完成将产生重大影响，使人们能够更深入地了解线粒体和氧化还原控制的细胞内过程，这些过程涉及我们对日常生活中遇到的环境压力的生物反应。