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

• 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
• 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

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 这个NRSA个人奖学金的总体目标是确定和研究新的治疗靶点 慢性阻塞性肺疾病（COPD）中由于香烟烟雾引起的上皮损伤，并促进 发展必要的技能，使候选人成为一个有效的物理学家，科学家。的 候选人和她的导师已经制定了一个严格的培训计划，将提供一个基础， 强大而成功的学术生涯。该提案涉及的基本生物过程， 线粒体和代谢调节参与吸烟相关的肺部疾病。慢性阻塞性 在美国，肺病（COPD）是第三大死亡原因，吸烟（CS）是第三大死亡原因。 导致疾病的原发性损伤。尽管正在进行哺乳动物研究，但没有新的生物靶点。 目前的治疗方法对疾病进展或死亡率的影响有限。我们正在利用 盘状网骨藻阿米巴与人类气道上皮细胞的比较研究 以确定新的途径和治疗策略，保护对CS。作为氧化损伤，CS导致 形态和代谢变化。我们已经发现，CS导致相关的生长表型 抑制Dictyosteoblastoma。此外，使用cDNA文库从约35，000个网骨藻中进行遗传选择， cDNA转化体暴露于香烟烟雾提取物（CSE），我们鉴定了腺嘌呤核苷酸移位酶 (ANT)作为对CS诱导的生长缺陷的保护。ANT是线粒体ADP/ATP转运蛋白 在线粒体代谢和能量平衡中起着积极作用。目前， 线粒体系统在保护肺上皮免受烟雾诱导的损伤中的作用仍不清楚。 该提案的长期目标是确定ANT保护免受 CS损伤和识别新的潜在治疗靶点。在目标1中，候选人将解读 人ANT对香烟烟雾（CS）后支气管上皮细胞（HBE）细胞活力和形态的影响。 这将通过分子表达工具，细胞成像和遗传学的组合来实现。 修饰这些蛋白质以评估细胞活力和结构。在目标2中，候选人将确定 代谢和炎症机制，通过这些机制人类ANT保护HBE免受CS损伤。这 将通过评估ANT引起的实时线粒体代谢和细胞因子变化来完成 HBE中的调制。这些实验的目的是利用哺乳动物和网骨藻模型， 剖析导致COPD中上皮失调的线粒体机制。实现这些目标 将阐明线粒体ANT在肺上皮细胞对CS的反应中的作用，作为一种新的保护机制， 在COPD中的路径。此外，这项工作将为检查新的治疗策略奠定基础 使用体内转化模型和FDA批准的药物库来治疗COPD。