Mitochondrial Genes as New Targets in the Protection of Airway Epithelial Cells Against Cigarette Smoke

线粒体基因作为保护气道上皮细胞免受香烟烟雾侵害的新靶标

基本信息

批准号：
9192357
负责人：
Corrine R Kliment
金额：
$ 5.32万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-01 至 2017-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9192357
关键词：
Actins Address Adenine Nucleotide Translocase Behavior Biological Assay Biological Models Biological Process Biology Carrier Proteins Cause of Death Cell Survival Cell model Cells Cellular Morphology Cellular Structures Chronic Obstructive Airway Disease Cigarette Complementary DNA Defect Development Dictyostelium Dictyostelium discoideum Disease Disease Progression Epithelial Epithelial Cells Epithelium Evaluation FDA approved Fellowship Foundations Future Genes Genus Hippocampus Glycolysis Goals Growth Homeostasis Human In Vitro Individual Inflammatory Injury Investigation Libraries Life Lung Lung diseases Mammalian Cell Measures Mediating Mentors Metabolic Metabolism Mitochondria Mitochondrial Proteins Modeling Molecular Myosin Type II National Research Service Awards Normal Cell Oxidative Phosphorylation Oxidative Stress Oxygen Consumption Pathogenesis Pathway interactions Patients Pharmaceutical Preparations Phenotype Physicians Play Post-Translational Protein Processing Production Regulation Research Role Scientist Smoker Structure Structure of parenchyma of lung Symptoms System Testing Therapeutic Time Tissues Training Translating United States Work Wound Healing base cDNA Library career cellular imaging cigarette smoke-induced cigarette smoking cigarette smoking comparative cytokine environmental tobacco smoke exposure experience fluorophore genetic manipulation genetic selection imaging genetics in vivo Model lung injury mitochondrial metabolism mortality mouse model new therapeutic target non-smoker novel novel therapeutic intervention novel therapeutics overexpression prevent protein function research study response skills small molecule therapeutic target tool

项目摘要

PROJECT SUMMARY The overall objectives of this NRSA individual fellowship are to identify and investigate new therapeutic targets for epithelial injury due to cigarette smoke in chronic obstructive lung disease (COPD), and to facilitate the development of essential skills that will allow the candidate to become an effective physician-scientist. The candidate and her mentors have constructed a rigorous training plan that will provide the foundation for a strong and successful academic career. The proposal addresses the fundamental biological processes of mitochondrial and metabolic regulation involved in cigarette smoking-related lung disease. Chronic obstructive pulmonary disease (COPD) is the 3rd leading cause of death in the US with cigarette smoke (CS) being the primary insult leading to disease. Despite ongoing mammalian research, no new biologic targets have been identified, and current therapies have limited effect on disease progression or mortality. We are leveraging the amoebozoan Dictyostelium discoideum to develop a comparative approach with human airway epithelial cells to identify new pathways and therapeutic strategies protective against CS. As an oxidative insult, CS results in morphologic and metabolic changes. We already found that CS leads to a relevant phenotype of growth suppression in Dictyostelium. Furthermore, using a cDNA library genetic selection from ~35,000 Dictyostelium cDNA transformants exposed to cigarette smoke extract (CSE), we identified adenine nucleotide translocase (ANT) as protective against CS-induced growth defects. ANT is a mitochondrial ADP/ATP transport protein that plays an active role in mitochondrial metabolism and energy homeostasis. Currently, the role of the ANT mitochondrial system in protecting the lung epithelium against cigarette-smoke induced injury remains unclear. The long-term goal of the proposal is to determine the mechanisms through which ANT is protective against CS injury and identify new potential therapeutic targets. In Aim 1, the candidate will decipher the effects of human ANT on cell viability and morphology in bronchial epithelial cells (HBEs) after cigarette smoke (CS). This will be accomplished by a combination of molecular expression tools, cell imaging, and genetic modification of these proteins to assess cell viability and structure. In Aim 2, the candidate will identify the metabolic and inflammatory mechanisms through which human ANT protects against CS injury in HBEs. This will be accomplished by evaluating real-time mitochondrial metabolism and cytokine changes due to ANT modulation in HBEs. The goal of these experiments is to leverage mammalian and Dictyostelium models to dissect the mitochondrial mechanisms leading to epithelial dysregulation in COPD. Completion of these aims will elucidate the role of the mitochondrial ANT in lung epithelium in response to CS, as a novel protective pathway in COPD. Furthermore, this work will lay groundwork for examination of new therapeutic strategies for COPD using translational in vivo models and an FDA-approved drug library.

项目摘要该NRSA个人奖学金的总体目标是识别和调查新的治疗目标由于慢性阻塞性肺疾病（COPD）中的香烟烟雾而引起的上皮损伤，并促进发展基本技能，使候选人成为有效的医师科学家。这候选人和她的导师制定了一项严格的培训计划，将为强大而成功的学术职业。该提案涉及的基本生物学过程与吸烟相关的肺部疾病涉及的线粒体和代谢调节。慢性阻塞性肺疾病（COPD）是美国的第三大死亡原因，香烟烟雾（CS）是主要侮辱导致疾病。尽管正在进行的哺乳动物研究，但没有新的生物学靶标确定的，当前的疗法对疾病进展或死亡率的影响有限。我们正在利用迪斯特尔迪斯特尔迪斯特尔（Dictyostelium discoideum）与人类气道上皮细胞开发一种比较方法确定针对CS的新途径和治疗策略。作为氧化损伤，CS导致形态学和代谢变化。我们已经发现CS导致相关的生长表型抑制dictyostelium。此外，使用〜35,000 dictyostelium的cDNA库遗传选择暴露于香烟烟雾提取物（CSE）的cDNA转化体，我们确定了腺嘌呤核苷酸易位酶（ANT）保护CS诱导的生长缺陷。蚂蚁是线粒体ADP/ATP转运蛋白这在线粒体代谢和能量稳态中起着积极作用。目前，蚂蚁的作用线粒体系统保护肺上皮免受烟雾诱发损伤的影响尚不清楚。该提案的长期目标是确定蚂蚁保护性的机制 CS损伤并确定新的潜在治疗靶标。在AIM 1中，候选人将破译香烟烟雾后支气管上皮细胞（HBE）中的细胞生存力和形态的人类蚂蚁（CS）。这将通过分子表达工具，细胞成像和遗传的组合来实现修饰这些蛋白质以评估细胞的生存力和结构。在AIM 2中，候选人将确定人类蚂蚁可以防止HBE中的CS损伤的代谢和炎症机制。这将通过评估蚂蚁引起的实时线粒体代谢和细胞因子变化来实现 HBES中的调制。这些实验的目的是利用哺乳动物和dictyostelium模型解剖导致COPD上皮失调的线粒体机制。这些目标的完成将阐明线粒体蚂蚁在肺上皮的CS中的作用，作为一种新型保护 COPD中的途径。此外，这项工作将为检查新的治疗策略奠定基础用于COPD使用体内模型和FDA批准的药物库的COPD。