Emphysema attenuation via modulation of macrophage NFKB and antiprotease activity

通过调节巨噬细胞 NFKB 和抗蛋白酶活性减轻肺气肿

• 批准号: 8117725
• 负责人: ANDREW A WILSON
• 金额: $ 12.96万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8117725
• 关键词: Affect; Alveolar Macrophages; Animals; Anti-Inflammatory Agents; Anti-inflammatory; Applications Grants; Award; Biological; Biological Assay; Biology; Blood; Cell Count; Cells; Chronic; Cigarette smoke-induced emphysema; Clinical; Critical Care; Development; Disease; Educational process of instructing; Elastases; Environment; Epithelial; Exposure to; Foundations; Goals; Hereditary Disease; Human; Immunology; In Vitro; Inflammation; Inflammatory; Infusion procedures; Inherited; Injury; Kinetics; Knowledge; Lead; Lentivirus Vector; Leukocyte Elastase; Life; Liquid substance; Lung; Lung Inflammation; Lung diseases; Mentors; Methodology; Methods; Modeling; Molecular Biology; Monitor; Mus; NF-kappa B; Nuclear; Nuclear Translocation; Obstruction; Pathogenesis; Patient Care; Patients; Physicians; Physiological; Play; Principal Investigator; Protease Inhibitor; Proteins; Pulmonary Emphysema; Pulmonology; Research; Research Personnel; Resolution; Respiratory physiology; Risk; Role; Scientist; Signal Transduction; Subfamily lentivirinae; System; Testing; Therapeutic; Time; Wild Type Mouse; Work; alpha 1-Antitrypsin; alpha 1-Antitrypsin Deficiency; attenuation; base; blood product; career; cell type; cellular transduction; cigarette smoke-induced; cigarette smoking; cytokine; gene therapy; healthy volunteer; in vivo; macrophage; novel; overexpression; p65; public health relevance; research study; response; skills; small hairpin RNA; symposium; tool; transcription factor

DESCRIPTION (provided by applicant): As a physician-scientist, my overarching career goal is to use the tools available to me in the research setting to work towards treatments for the lung diseases that I encounter in the clinical arena. I plan to become an independent principal investigator in an academic division of pulmonary medicine and to spend approximately 80% of my time pursuing scientific research. Patient care and teaching responsibilities as a pulmonary and critical care attending will occupy the remainder of my time and help to keep me focused on the diseases that drive our research efforts. I plan to use the K08 award to build upon my initial work on lentiviral manipulation of alveolar macrophages and the role that these cells play in emphysema pathogenesis. During this time, I plan to develop my research career by: 1) laying an educational foundation in immunology through a combination of formal coursework and regular attendance at a weekly pulmonary immunology conference to discuss my research and that of others; 2) expanding my technical repertoire to include additional skills central to a career in molecular biology research; 3) using the assets acquired in numbers 1 and 2 above to extend and broaden my work to date; and 4) transitioning from a mentored research environment to one of independent scientific inquiry. Patients with alpha-1 antitrypsin (AAT) deficiency have decreases in circulating blood and lung epithelial lining fluid AAT that ultimately lead to panacinar emphysema, particularly in the setting of cigarette smoke exposure. Current therapy for patients with impaired lung function resulting from AAT deficiency is replacement via weekly infusions of AAT from pooled human plasma.4 This treatment is expensive, inefficient (only a small fraction of the infused AAT reaches the lung),5 and involves a lifetime of exposure to human blood products and the risks that accompany such exposure. A comprehensive understanding of AAT biology is lacking and has been a major barrier to development of new treatments for AAT deficient patients. Although it has long been known that AAT functions to inactivate neutrophil elastase in the lung, it has only recently been recognized to have a variety of alternative and significant biological effects. In particular, the effects that AAT exerts on signaling of the key transcription factor nuclear factor-kB (NF-kB) are not well understood. The goals of this proposal are first, to better understand the relationship between AAT and NF-kB to help further our knowledge of emphysema pathogenesis; and second, to test a potential gene therapy for AAT deficiency based on local expression of the normal human AAT protein in the alveolar macrophage (AM), the cell type most implicated in lung destruction in AAT deficient patients. To accomplish these goals, the investigators have developed a new tool allowing tracking of NF-kB activity in live animals, something that was not previously possible. The experiments outlined further develop this tool and then use it to help test whether anti-inflammatory effects exerted by AAT in the lung are accompanied by suppression of NF-:B translocation and additionally whether AAT deficiency is characterized by chronic activation of NF-:B signaling. They test the effects of manipulating NF-:B signaling in the setting of cigarette smoke exposure. They then proceed to test whether or not life-long overexpression of AAT in resident AMs ameliorates emphysema via anti-inflammatory effects characterized by suppression of macrophages NF-:B activation in normal or AAT deficient mice. PUBLIC HEALTH RELEVANCE: Alpha-1 antitrypsin (AAT) deficiency is one of the most common hereditary lung diseases world-wide. Findings helping to explain how AAT protects the lung in the setting of injury are therefore likely to have great significance. Any novel treatment capable of affecting progression of emphysema will similarly have a significant long-term clinical impact.

描述（由申请人提供）：作为一名医生，科学家，我的首要职业目标是利用我在研究环境中可用的工具，致力于治疗我在临床竞技场中遇到的肺部疾病。我计划成为一名独立的首席研究员，在肺部医学的学术部门，花大约80%的时间从事科学研究。作为一名肺部和重症监护主治医师，病人护理和教学职责将占用我剩余的时间，并有助于让我专注于推动我们研究工作的疾病。我计划利用K 08奖来建立我对肺泡巨噬细胞的慢病毒操作的初步工作，以及这些细胞在肺气肿发病机制中的作用。在此期间，我计划通过以下方式发展我的研究事业：1）通过正式课程和定期参加每周肺免疫学会议来讨论我的研究和其他人的研究，从而奠定免疫学的教育基础; 2）扩展我的技术技能，包括分子生物学研究职业生涯中的其他技能; 3）利用上面第1和第2项中获得的资产来扩展和拓宽我迄今为止的工作; 4）从指导研究环境过渡到独立的科学探究环境。α-1抗胰蛋白酶（AAT）缺乏的患者循环血液和肺上皮衬里液AAT减少，最终导致全腺泡肺气肿，特别是在香烟烟雾暴露的情况下。目前对AAT缺乏导致肺功能受损的患者的治疗是通过每周输注来自汇集的人血浆的AAT进行替代。4这种治疗昂贵、低效（只有一小部分输注的AAT到达肺部），5并且涉及终生暴露于人血液制品以及伴随这种暴露的风险。缺乏对AAT生物学的全面了解，这一直是AAT缺陷患者开发新疗法的主要障碍。尽管人们早就知道AAT在肺中具有抑制中性粒细胞弹性蛋白酶的功能，但直到最近才认识到它具有多种替代性和显著的生物学效应。特别是，AAT对关键转录因子核因子-kB（NF-kB）信号传导的影响尚未得到充分理解。该建议的目标是，首先，更好地了解AAT和NF-κ B之间的关系，以帮助进一步了解我们的肺气肿发病机制;第二，测试一个潜在的基因治疗AAT缺乏症的基础上，在肺泡巨噬细胞（AM），细胞类型的正常人AAT蛋白的局部表达最牵连在肺破坏AAT缺乏症患者。为了实现这些目标，研究人员开发了一种新的工具，可以跟踪活体动物中的NF-kB活性，这在以前是不可能的。概述的实验进一步开发了该工具，然后使用它来帮助测试AAT在肺中发挥的抗炎作用是否伴随NF-：B易位的抑制，以及AAT缺乏是否以NF-：B信号传导的慢性激活为特征。他们测试了在暴露于香烟烟雾的情况下操纵NF-：B信号的效果。然后，他们继续测试在正常或AAT缺陷小鼠中，常驻AM中AAT的终身过表达是否通过以抑制巨噬细胞NF-：B活化为特征的抗炎作用来改善肺气肿。 公共卫生相关性：α-1抗胰蛋白酶（AAT）缺乏症是世界范围内最常见的遗传性肺病之一。因此，有助于解释AAT如何在损伤情况下保护肺的发现可能具有重要意义。任何能够影响肺气肿进展的新型治疗方法都将同样具有显著的长期临床影响。