Emphysema attenuation via modulation of macrophage NFKB and antiprotease activity

描述（由申请人提供）：作为一名内科科学家，我的总体职业目标是使用我在研究环境中可用的工具来研究我在临床领域遇到的肺部疾病的治疗方法。我计划成为肺医学学术部门的独立首席研究员，并将大约80%的时间用于科学研究。作为一名肺病和重症监护主治医生，病人护理和教学职责将占据我剩余的时间，并帮助我专注于推动我们研究工作的疾病。我计划利用K08奖来建立我在慢病毒操纵肺泡巨噬细胞和这些细胞在肺气肿发病机制中所起作用的初步工作。在此期间，我计划通过以下方式发展我的研究事业：1)通过正规的课程学习和定期参加每周一次的肺免疫学会议来讨论我和他人的研究，奠定免疫学的教育基础；2)扩大我的技能范围，包括分子生物学研究职业生涯的核心技能；3)利用上述第1和第2项所获得的资产来扩展和拓宽我迄今为止的工作；4)从受指导的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内层液AAT减少，最终导致panacinar肺气肿，特别是在香烟烟雾暴露的情况下。目前对因AAT缺乏而导致肺功能受损的患者的治疗是通过每周从汇集的人血浆中输注AAT来替代这种治疗费用昂贵，效率低下（只有一小部分注入的AAT到达肺部），5并且涉及终生接触人类血液制品以及伴随这种接触的风险。缺乏对AAT生物学的全面了解，这已经成为开发针对AAT缺陷患者的新疗法的主要障碍。虽然人们早就知道AAT具有肺中性粒细胞弹性酶失活的功能，但直到最近才认识到它具有多种替代和显着的生物学作用。特别是，AAT对关键转录因子核因子- kb （NF-kB）信号转导的影响尚不清楚。本研究的目标是：首先，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的发病机制；第二，基于肺泡巨噬细胞（AM）中正常人类AAT蛋白的局部表达，测试AAT缺乏症的潜在基因治疗方法，AM是AAT缺乏症患者肺破坏最相关的细胞类型。为了实现这些目标，研究人员开发了一种新的工具，可以跟踪活体动物的NF-kB活性，这在以前是不可能的。实验概述了进一步发展这一工具，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着NF-:B易位的抑制，以及AAT缺乏是否以NF-:B信号的慢性激活为特征。他们测试了在吸烟环境下操纵NF-:B信号的效果。然后，他们进一步测试在正常或AAT缺陷小鼠中，AAT的终身过度表达是否通过抑制巨噬细胞NF-:B活化的抗炎作用来改善肺气肿。

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.