Mannose binding lectin-dependent complement activation in emphysema

项目总结 香烟烟雾激活补体级联导致肺不育的机制 炎症和COPD进展尚不完全清楚。考虑到网络的关键作用 病原体诱导的炎症中的补体，选择性抑制甘露糖结合凝集素(MBL) 补体途径可能导致CS诱导的炎症和肺气肿样空隙减少 放大，而不是不加区别地抑制补体对病原体的反应。在目标1中，我们建议 探讨CS致肺损伤的新机制；重点研究甘露糖结合凝集素 相关丝氨酸蛋白酶-2(MASP-2)是MBL途径中的中心蛋白酶，是诱导 接触CS时的炎症和肺气肿。在目标2中，我们将研究α-1抗胰蛋白酶、 主要的丝氨酸蛋白酶抑制剂，结合并抑制MASP-2。我的建议针对的是与临床相关的 质疑通过抑制MASP-2选择性靶向补体级联反应是否能改善肺部 CS暴露相关小鼠模型中炎症和肺气肿的发生。我们的动物研究是 补充测量MASP-2水平和活性的样本来自活跃吸烟者和 没有慢性阻塞性肺病。 拟议课程和实验的设计和实施将帮助我获得 补体生物学、酶学方面的专业知识，以及完善我的肺部体视学技能。我将在新的训练 差示温热电泳法、等温滴定量热法和荧光共振等技术 能量转移，FRET-荧光寿命成像显微镜，胶片研究α-1抗胰蛋白酶与 MASP-2。这些技巧和新的研究集中在补体系统在CS诱导的不孕中的作用 炎症将使我成为一名独立的研究员，从事不同于我的 导师，彼得拉什医生。我的委员会成员和合作者处于战略地位，可以协助 在完成这项建议方面，我谨此致谢。国家犹太人健康环境，超凡的专业知识 指导团队，以及Petrache博士与 委员会，将确保一个多方面和培育的环境，以促进我向独立的过渡。 该项目的完成将提供令人信服的实验证据，证明利用MASP-2抑制 蛋白水解酶抑制剂可减轻肺气肿小鼠模型的炎症和肺损伤 作为人类COPD疾病的下一代治疗药物。

Project summary The mechanisms by which cigarette smoke (CS) activates the complement cascade to cause lung sterile inflammation and COPD progression are not completely understood. Considering the critical role of complement in pathogen-induced inflammation, selective inhibition of the mannose-binding lectin (MBL) complement pathway may result in decreased CS-induced inflammation and emphysema-like airspace enlargement without an indiscriminate inhibition of complement's response to pathogens. In Aim 1 we propose to investigate a novel mechanism of CS-induced lung injury; focusing on whether mannose-binding lectin associated serine protease-2 (MASP-2), the central protease in the MBL pathway, is necessary to induce inflammation and emphysema during CS exposure. In Aim 2 we will investigate whether alpha-1 antitrypsin, a major serine protease inhibitor, binds and inhibits MASP-2. My proposal addresses the clinically relevant question whether selective targeting of complement cascade via MASP-2 inhibition ameliorates lung inflammation and emphysema development in relevant murine models of CS exposure. Our animal studies are complemented by measurements of MASP-2 levels and activity in samples from active smokers with and without COPD. The design and implementation of the proposed coursework and experiments will assist me in gaining expertise in complement biology, enzymology, and perfect my skills in lung stereology. I will train in new techniques, such as miscroscale thermophoresis, isothermal titration calorimetry, and fluorescence resonance energy transfer, FRET - fluorescence lifetime imaging microscopy, FLIM to study alpha-1 antitrypsin binding to MASP-2. These skills and the new research focus on the role of complement system in CS-induced sterile inflammation will allow me to become an independent investigator in a different area of research than my mentor, Dr. Petrache. My committee members and collaborators are strategically positioned to assist me in completing this proposal. National Jewish Health environment, the extraordinary expertise of the mentoring team, and the track record of fruitful collaborations between Dr. Petrache and members of the committee, will ensure a multifaceted and nurturing setting to facilitate my transition to independence. Completion of this project will provide compelling experimental evidences that MASP-2 inhibition using protease inhibitors ameliorates inflammation and lung injury in murine models of emphysema and it can be harnessed as next generation therapeutics in human COPD disease.