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起重要作用。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，这些酶通过水解胞外蛋白参与炎症过程。其具体目的是：1.探索DPPI的结构与功能关系；2.鉴定正常肺组织和炎症肺组织中表达DPPI的细胞；3.鉴定DPPI的底物特异性；4.鉴定肥大细胞中发现的一种新的60 kDa半胱氨酸蛋白酶。目的：1.在重组表达的DPPI中对ProDPPI的蛋白降解过程进行研究。用免疫组织化学和原位杂交法检测DPPI在正常和炎症犬肺组织中的表达。4.将研究DPPI对胞外肽和蛋白质的水解性。5.将在DPPI基因敲除小鼠中研究肥大细胞的功能特征和蛋白降解活性。6.纯化并鉴定了一种新型肥大细胞半胱氨酸蛋白酶。沃尔特斯医生已经完成了内科、肺部和重症监护医学方面的培训。他表现出了对学术医学事业的坚定承诺，并对研究肺部疾病的病理生理学感兴趣。研究培训计划包括：密集的实验室体验、教学课程、每周研讨会和期刊俱乐部。研究培训将由一个顾问委员会监督，该委员会包括赞助商(考伊博士)和半胱氨酸蛋白酶专家(麦克克罗博士)、免疫组织病理学专家(麦克唐纳博士)和肺部疾病专家(马泰博士)，他们将指导候选人发展为肺部研究的独立研究员。

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.