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: 8501652
• 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/8501652
• 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.

描述（由申请人提供）：作为医师科学家，我的总体职业目标是使用研究环境中可用的工具来努力治疗我在临床领域遇到的肺部疾病。我计划成为肺医学学术部门的独立首席研究员，并花费大约80％的时间从事科学研究。患者护理和教学责任作为肺部和重症监护，将占据我的剩余时间，并帮助我专注于推动我们研究工作的疾病。我计划使用K08奖，以我对肺泡巨噬细胞操纵的最初工作以及这些细胞在肺气肿发病机理中的作用。在此期间，我计划通过以下方式发展自己的研究职业：1）通过正式课程和定期参加每周一次的肺部免疫学会议的结合，为我的研究和其他研究奠定免疫学教育基金会； 2）扩大我的技术曲目，包括分子生物学研究职业中心的其他技能； 3）使用上面数字1和2中获得的资产来扩展和扩大我的工作； 4）从指导的研究环境过渡到独立科学探究之一。 α-1抗胰蛋白酶（AAT）缺乏症患者的循环血液和肺上皮衬里AAT的降低，最终导致了灵活的肺气肿，尤其是在香烟烟雾的情况下。 AAT缺乏症引起的肺功能受损的患者的当前治疗是通过每周从合并的人血浆中注入AAT的替代。4这种疗法昂贵，效率低下（只有一小部分注入的AAT到达肺），5），5，涉及对人类血液产物的一生暴露于人类血液和伴随这种暴露的风险。缺乏对AAT生物学的全面理解，并且是为AAT缺乏患者开发新治疗方法的主要障碍。尽管早就知道AAT在肺中灭活性嗜中性粒细胞弹性酶的作用，但直到最近才被认为具有多种替代性和重要的生物学作用。特别是，AAT对关键转录因子核因子-KB（NF-KB）的信号传导的影响尚不清楚。该提案的目标首先是为了更好地了解AAT和NF-KB之间的关系，以帮助我们进一步了解肺气肿发病机理；其次，基于肺泡巨噬细胞中正常人AAT蛋白的局部表达测试潜在的基因治疗，以使AAT缺乏症（AM），这是对AAT缺乏患者的肺部破坏最大的细胞类型。为了实现这些目标，调查人员开发了一种新工具，允许跟踪活动物中的NF-KB活动，这是以前无法的。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在肺中施加的抗炎作用是否伴随着NF-：B易位的抑制以及AAT缺乏症是否以NF-：B信号的慢性激活为特征。他们测试操纵NF-：B信号在烟烟暴露中的作用。然后，他们继续测试居民AM中AAT的终生过表达是否通过抑制巨噬细胞NF-的特征在于正常或AAT缺陷小鼠中的巨噬细胞NF-激活。