PATHOBIOLOGY OF BRONCHIAL TISSUE KALLIKREIN

描述 (改编自申请人的摘要)本提案的目的是 提供Rosanna Forteza，医学博士，助理教授 迈阿密大学(UM)肺部和重症监护医学 在酶学和分子生物学方面的深入研究培训，这样她就可以 成为哮喘的独立研究人员和积极贡献者 该部门的研究小组。哮喘研究项目正在积极开展 参与从哮喘的分子和细胞基础的一系列研究 以其生理和疾病的临床表现。这 该项目将利用两个基地：罗森斯泰尔医学大楼 密歇根大学校园(细胞和分子研究)和西奈山医学 中心(生理学和药理学研究)。亚当·万纳博士，首席 该部门将与威廉博士一起担任Forteza博士的主要赞助人 M.亚伯拉罕和格雷戈里·康纳担任次要赞助商。Forteza博士 将研究支气管组织激肽释放酶在呼吸道中的作用。 与三位赞助商共同完成的初步研究使Dr。 Forteza提出的假设是，呼吸道高反应性取决于 支气管组织激肽释放酶上调及其后代的研究 激肽可能至少部分地负责维持 呼吸道炎症，易导致呼吸道高反应性。至 更好地了解哮喘患者支气管组织激肽释放酶的调节 航空公司，已经确定了两个具体目标。在特定目标1中，绵羊 将对支气管激肽释放酶进行纯化，并对其特性进行研究 结构、酶活性和对抑制剂的敏感性。在……里面 此外，Forteza博士还将从绵羊的气管中克隆激肽释放酶基因 图书馆。纯化的激肽释放酶将用于生产抗血清 定量分析(ELISA、RIA)和免疫组织化学，而 绵羊激肽释放酶基因将作为研究基因表达的探针 在正常和发炎情况下。在《特定目标2》中，Forteza博士将 研究支气管组织的基因表达、合成和释放 激肽释放酶在正常情况下，引起呼吸道炎症后 抗原挑战，并在此抗原修饰后诱导 糖皮质激素引起的炎症。由于研究的性质， 这些实验将在已建立的绵羊模型中进行 抗原诱导的呼吸道高反应性。总的来说，这些研究 应有助于提高对支气管组织激肽释放酶和 可能会带来新的治疗方式。除了她的研究 经验，Forteza博士的培训将辅之以两个说教 细胞和分子生物学课程。她在整个课程中的进步 将在包括Goal在内的领域通过书面评估来监测赠款的使用情况 树立、贯彻和全面的责任意识。这 计划将为Forteza博士提供适当的专业知识，以开始 富有成效的独立学术生涯。

DESCRIPTION (Adapted from the applicant's abstract) The purpose of this proposal is to provide Rosanna Forteza, M.D., an assistant professor in the Division of Pulmonary and Critical Care Medicine at the University of Miami (UM) with in depth research training in enzymology and molecular biology, so that she can become an independent researcher and active contributor to the asthma research group in the division. The asthma research program is actively involved in studies ranging from the molecular and cellular basis of asthma to its physiology and the clinical manifestations of the disease. This project will utilize two bases: the Rosensteil Medical Science Building on the UM campus (cellular and molecular studies) and Mount Sinai Medical Center (physiologic and pharmacologic studies). Dr. Adam Wanner, chief of the division, will serve as Dr. Forteza's primary sponsor, with Drs. William M. Abraham and Gregory Conner serving as secondary sponsors. Dr. Forteza will study the role of bronchial tissue kallikrein in the airways. Preliminary studies done in conjunction with the three sponsors led Dr. Forteza to formulate the hypothesis that airway hyperresponsiveness depends on upregulation of bronchial tissue kallikrein and the subsequent generation of kinins which may be responsible, at least in part, for the maintenance of airway inflammation, which predisposes to airway hyperresponsiveness. To better understand the regulation of bronchial tissue kallikrein in the airways, two specific aims have been identified. In Specific Aim 1, ovine bronchial kallikrein will be purified and characterized with respect to its structure, enzymatic activity, and susceptibility to inhibitors. In addition, Dr. Forteza will clone kallikrein cDNA from a sheep tracheal library. Purified kallikrein will be used to produce antisera for quantitative analysis (ELISA, RIA) and immunohistochemistry, whereas the sheep kallikrein cDNA will be used as a probe for studying gene expression under normal and inflamed conditions. In Specific Aim 2, Dr. Forteza will investigate gene expression, synthesis and release of bronchial tissue kallikrein under normal conditions, after airway inflammation caused by antigen challenge, and after modification of this antigen-induced inflammation by glucocorticosteroids. Because of the nature of the studies, these experiments will be carried out in the well-established sheep model of antigen-induced airway hyperresponsiveness. Collectively, these studies should help improve understanding of bronchial tissue kallikrein and possibly lead to new therapeutic modalities. In addition to her research experience, Dr. Forteza's training will be supplemented by two didactic courses in cell and molecular biology. Her progress throughout the course of the grant will be monitored by written evaluation in areas including goal setting, follow-through, and overall awareness of responsibility. This program will provide Dr. Forteza with the appropriate expertise to begin a productive independent academic career.