Mannose binding lectin-dependent complement activation in emphysema

肺气肿中甘露糖结合凝集素依赖性补体激活

• 批准号: 10310458
• 负责人: Karina Serban
• 金额: $ 15.87万
• 依托单位: NATIONAL JEWISH HEALTH
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-12-17 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10310458
• 关键词: Address; Animals; Antigen-Antibody Complex; Apical; Area; Binding; Biological Assay; Biological Markers; Biology; Calorimetry; Cause of Death; Cell surface; Cells; Chronic Obstructive Pulmonary Disease; Co-Immunoprecipitations; Code; Collaborations; Committee Members; Complement; Complement Activation; Data; Deposition; Development; Disease; Disease Progression; Energy Transfer; Ensure; Environment; Enzymatic Biochemistry; Enzyme-Linked Immunosorbent Assay; Epithelial; Epithelial Cells; Event; Fluorescence; Fluorescence Resonance Energy Transfer; Health; Human; In Vitro; Incidence; Individual; Infection; Inflammation; Kidney; Lead; Lectin; Link; Liquid substance; Lung; Lung diseases; MASP2 gene; Mannose Binding Lectin; Measurement; Measures; Mentors; Mus; Myocardial Ischemia; Pathogenicity; Pathway interactions; Peptide Hydrolases; Phase; Point Mutation; Positioning Attribute; Predisposition; Protease Inhibitor; Protein Isoforms; Pulmonary Emphysema; Pulmonary Inflammation; Reporting; Research; Research Personnel; Role; Sampling; Serine; Serine Protease; Serine Proteinase Inhibitors; Serum; Severity of illness; Signal Transduction; Smoke; Smoker; Smoking; Sterility; Structure of parenchyma of lung; Surface; Techniques; Testing; Tissues; Titrations; Training; Translating; Wild Type Mouse; airway epithelium; alpha 1-Antitrypsin; base; cell injury; cigarette smoke; cigarette smoke-induced; cigarette smoke-induced inflammation; cigarette smoke-induced lung injury; cigarette smoking; clinically relevant; complement pathway; complement system; design; early onset; experimental study; exposure to cigarette smoke; fluorescence lifetime imaging; follower of religion Jewish; improved; in vivo; inhibitor; lung injury; mannose-binding protein-associated serine proteases; mortality; mouse model; novel; novel therapeutics; pathogen; recruit; response; skills; tissue injury; translational study

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.

项目摘要 香烟烟雾（CS）激活补体级联反应导致肺不育的机制 炎症和COPD进展尚未完全了解。考虑到联合国的关键作用， 补体在病原体诱导的炎症中，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域减少 在没有不加选择地抑制补体对病原体的反应的情况下，在目标1中，我们提出 探讨CS诱导肺损伤的新机制，重点研究甘露糖结合凝集素是否 相关丝氨酸蛋白酶-2（MASP-2），MBL途径中的中心蛋白酶，是诱导细胞凋亡所必需的。 炎症和肺气肿。在目标2中，我们将研究α-1抗胰蛋白酶， 主要丝氨酸蛋白酶抑制剂，结合并抑制MASP-2。我的提案涉及临床相关问题 质疑通过MASP-2抑制选择性靶向补体级联是否改善肺 炎症和肺气肿发展。我们的动物研究是 补充的是测量来自患有和患有以下疾病的活跃吸烟者的样品中的MASP-2水平和活性， 没有COPD。 拟议的课程和实验的设计和实施将有助于我获得 补充生物学、酶学方面的专业知识，完善我在肺体视学方面的技能。我将在新的 微尺度热泳、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，FLIM研究α-1抗胰蛋白酶结合 MASP-2。这些技术和新的研究集中在补体系统在CS诱导的不育中的作用 炎症将使我成为一个独立的研究人员在一个不同的研究领域比我的 导师彼得拉奇博士我的委员会成员和合作者处于战略地位， 我来完成这个提案。国家犹太健康环境，非凡的专业知识， 指导团队，以及Petrache博士与 委员会将确保一个多方面和有利的环境，以促进我向独立过渡。 该项目的完成将提供令人信服的实验证据，表明使用MASP-2抑制剂可以抑制MASP-2。 蛋白酶抑制剂改善肺气肿小鼠模型中的炎症和肺损伤， 作为人类COPD疾病的下一代疗法。