Pathogenic Exosomes in COPD

项目摘要 这个R35计划的论点是，免疫细胞来源的“致病性外泌体”是 慢性阻塞性肺疾病（COPD）的主要参与者，他们的作用可以是 在吸烟小鼠外泌体转移模型中建模，以获得对 这种疾病可以反过来应用于COPD。该R35计划处于以下领域的前沿： 创新发现了α-1抗胰蛋白酶（α-1AT）抗性中性粒细胞的存在， 弹性蛋白酶（NE）+中性粒细胞（PMN）衍生的外泌体。为此，该方案 已经挑战了蛋白酶活性仅仅是溶液相的概念， 细胞外基质（ECM）蛋白水解活性通过蛋白酶附着到细胞外基质上而大大增强。 细胞外囊泡（EV）表面。因此，我们：i）描述似乎是第一个EV 可以将COPD样表型从人类转移到小鼠; ii）阐明一种新的机制 蛋白酶通过其逃避抗蛋白酶失活，导致ECM降解和细胞凋亡。 死亡通过受体相互作用蛋白激酶3（RIPK 3）; iii）描述一种新的模型， a从香烟烟雾（CS）暴露小鼠通过免疫细胞- iv）使用小鼠模型发现新的CS诱导的保护性外泌体; 抗外泌体损伤的机制以及外泌体自我繁殖的机制; v）发现外泌体损伤的新的治疗靶点; 更好地了解COPD患者和吸烟者的疾病。 这项研究计划的另一个非常重要的方面是培训/指导。在 过去十年，其中包括PI的就职R35赠款，PI已经或目前 导师两名博士生，五名医学生，一名MD/博士生，六名K获奖者，四名 博士后实习生，一个肺科研究员，和一个心脏病学研究员。PI目前指导 七名初级教员总的来说，我们的计划是多学科-就业 化学，生物化学，分子和细胞生物学，动物生理学等-和翻译 将基础科学家与内科科学家配对。我们的团队和学员履行NHLBI/NIH 使命是通过包括非洲裔美国人、拉丁裔、亚裔和高加索人来实现多样性 成员我们的计划通过以下方式实现了NHLBI战略愿景的所有四个目标：a）阐明 人类生物学的新的外泌体方面; B）通过新的治疗方法减少人类疾病 c）开发工作队伍和动物模型 外泌体资源和; d）将外泌体研究转化为人类疾病。

PROJECT SUMMARY The thesis of this R35 Program is that immune cell derived “pathogenic exosomes” are key players in chronic obstructive pulmonary disease (COPD) and that their role can be modeled in a smoking mouse model of exosome transfer to attain novel understanding of the disease which can in turn be applied to COPD. This R35 Program is at the cutting edge of innovation having discovered the existence of alpha-1 antitrypsin (α-1AT) resistant neutrophil elastase (NE)+ neutrophil (PMN)-derived exosomes in COPD patients. In doing so, the Program has challenged the concept that protease activity is solely solution phase and shown extracellular matrix (ECM) proteolytic activity is hugely enhanced by protease attachment to the extracellular vesicle (EV) surface. Consequently, we: i) describe what appears to be the first EV that can transfer a COPD-like phenotype from humans to mice; ii) elucidate a new mechanism by which proteases escape anti-protease inactivation, leading to ECM degradation and cell death via receptor-interacting protein kinase 3 (RIPK3); iii) describe a new model that transfers a COPD phenotype from cigarette smoke (CS) exposed mice to naïve mice via immune cell- derived exosomes; iv) use the mouse model to discover a new CS induced protective mechanism against exosomal damage as well as a mechanism for exosomal self-propagation; v) uncover new therapeutic targets for exosomal damage and; vi) translate the new findings to better understand disease in COPD patients and smokers. Another highly significant aspect of this research Program is training/mentoring. In the past ten years, which includes the PI's inaugural R35 grant, the PI has been or is currently mentor to two PhD students, five medical students, an MD/PhD student, six K awardees, four postdoctoral trainees, a pulmonary fellow, and a cardiology fellow. The PI currently mentors seven junior faculty members. Overall, our Program is both multidisciplinary – employing chemistry, biochemistry, molecular and cell biology, animal physiology, etc. – and translational in pairing basic scientists with physician scientists. Our team and trainees fulfill the NHLBI/NIH mission to have diversity by including African American, Latina, Asian, and Caucasian members. Our program fulfills all four goals of the NHLBI Strategic Vision by: a) elucidating a new exosomal aspect of human biology; b) reducing human disease by new therapeutic development against the exosomal pathway; c) developing a work force and an animal model exosome resource and; d) translating the exosome research to human disease.