MAST CELL CYSTEINE PROTEASES IN LUNG INFLAMMATION

肥大细胞半胱氨酸、金属蛋白酶和丝氨酸蛋白酶可能在炎症性肺部疾病的发病机制中发挥重要作用。这项建议的长期目标是确定肥大细胞中存在的半胱氨酸蛋白酶，并表征它们在肥大细胞依赖性炎症中的作用。我们假设组织肥大细胞是二肽基肽酶I （DPPI）和一种新发现的60 kda半胱氨酸蛋白酶的主要来源，这些蛋白酶通过水解细胞外蛋白参与炎症反应。具体目标是：1.；探讨DPPI的结构-功能关系；2. 鉴定正常和炎症肺组织中表达DPPI的细胞；3. 3 .表征DPPI的底物特异性；描述肥大细胞中发现的一种新的60 kDa半胱氨酸蛋白酶。要实现这些目标：将在重组表达的DPPI中研究proDPPI的蛋白水解过程。2. 用突变前肽表达的DPPI将检测其二肽基肽酶和内源性蛋白酶活性。通过免疫组织化学和原位杂交检测正常和炎症狗肺中特定细胞类型DPPI的表达。4. 将研究DPPI水解细胞外肽和蛋白质的能力。5. 将在DPPI敲除小鼠中研究肥大细胞的功能特征和蛋白水解活性。6. 一种新的肥大细胞半胱氨酸蛋白酶将被纯化和表征。Wolters博士完成了内科、肺部和重症监护医学的培训。他坚定地致力于医学学术事业，并对研究肺部疾病的病理生理学感兴趣。研究训练计划包括：密集的实验室体验、教学课程、每周研讨会和期刊俱乐部。研究培训将由咨询委员会监督，该委员会包括发起人（Dr. Caughey）和半胱氨酸蛋白酶（Dr. McKerrow）、免疫组织病理学（Dr. McDonald）和肺部疾病（Dr. Matthay）方面的专家，他们将指导候选人发展成为肺部研究的独立研究者。

Mast cell cysteine, metalloprotease, and serine proteases may play a significant role in the pathogenesis of inflammatory lung diseases. The long-term objective of this proposal is to identify the cysteine proteases present in mast cells, and to characterize their roles in mast cell dependent inflammation. We hypothesize that tissue mast cells are a major source of dipeptidyl peptidase I (DPPI) and a newly identified 60-kDa cysteine protease and that these proteases participate in inflammation by hydrolyzing extracellular proteins. Specific aims are : 1. To explore structure-function relationships of DPPI; 2. To identify cells expressing DPPI in normal and inflamed lung tissues; 3. To characterize substrate specificity of DPPI, and 4. To characterize a nove1 60 kDa cysteine protease found in mast cells. To achieve these goals: 1. Proteolytic processing of proDPPI will be studied in recombinantly expressed DPPI. 2. DPPI's expressed with mutated propeptides will be assayed for their dipeptidyl peptidase and endoprotease activity, 3. Expression of DPPI by specific cell types in normal and inflamed dog lungs will be determined by immunohistochemistry and in situ hybridization. 4. DPPI will be studied for its ability to hydrolyze extracellular peptides and proteins. 5. Mast cell functional characteristics and proteolytic activities will be studied in DPPI knockout mice. 6. A novel mast cell cysteine protease will be purified and characterized. Dr. Wolters has completed training in internal medicine and pulmonary and critical care medicine. He has demonstrated a firm commitment to a career in academic medicine and an interest in studying the pathophysiology of lung diseases. The research training plan includes: an intensive laboratory experience, didactic coursework, weekly seminars, and journal clubs. The research training will be overseen by an advisory committee including the sponsor (Dr. Caughey) and experts on cysteine proteases (Dr. McKerrow), immunohistopathology (Dr. McDonald), and pulmonary diseases (Dr. Matthay), who will guide the candidates development into an independent investigator in pulmonary research.