PATHOBIOLOGY OF BRONCHIAL TISSUE KALLIKREIN

描述 （改编自申请人的摘要）本建议书的目的是 提供罗莎娜·福泰萨医学博士一个助理教授在司 肺和重症监护医学在迈阿密大学（UM）与 在酶学和分子生物学方面的深入研究培训，使她能够 成为一个独立的研究人员和积极的贡献者哮喘 研究组在司。 哮喘研究计划正在积极地 从哮喘的分子和细胞基础， 生理学和临床表现的联系 这 该项目将利用两个基地：罗森斯泰尔医学科学大楼， UM校园（细胞和分子研究）和西奈山医学中心 中心（生理学和药理学研究）。 Adam Wanner博士， 该部门将作为Forteza博士的主要赞助商， 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气道中的作用。 与三个赞助商一起进行的初步研究导致了Dr. Forteza提出了气道高反应性依赖于 对支气管组织激肽释放酶的上调和随后的 激肽可能是负责，至少部分，为维护 气道炎症，易导致气道高反应性。 到 更好地了解支气管组织激肽释放酶的调节， 航空公司，已确定两个具体目标。 在目标特定1中，绵羊 支气管激肽释放酶将被纯化并就其特性进行表征。 结构、酶活性和对抑制剂的敏感性。 在 此外，Forteza博士将从绵羊气管克隆激肽释放酶cDNA 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）和免疫组化，而 绵羊激肽释放酶cDNA将被用作研究基因表达的探针 在正常和发炎的情况下。 在《特定目标2》中，Forteza博士将 研究支气管组织基因表达、合成和释放 激肽释放酶在正常情况下，后气道炎症引起的 抗原攻击，并且在修饰该抗原诱导的 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， 抗原诱导的气道高反应性。 总的来说，这些研究 应该有助于提高对支气管组织激肽释放酶的理解， 可能会带来新的治疗方法。 除了她的研究 根据经验，Forteza博士的培训将由两个教学补充 细胞和分子生物学课程。 她在整个课程中的进步 赠款的使用将通过书面评估进行监测，包括目标 设定、贯彻和全面的责任意识。 这 该计划将为Forteza博士提供适当的专业知识，以开始一个 独立的学术生涯。 同行评审员的评论 简历和其他开头部分的简要说明是 对小组讨论的最终结果的权威性陈述。 的 所附评论是由指定的审查员编写的， 应用程序. 所有评审员都对申请进行了讨论和评分 礼物 因此，评注可能不一定反映 作者在小组讨论结束时也没有最终的多数意见 最后一个

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. PEER REVIEWERS' COMMENTS The resume and other initial sections of the summary statement are the authoritative representation of the final outcome of group discussion. The comments attached were prepared by the reviewers assigned to this application. The application was discussed and scored by all reviewers present. Thus, the commentaries may not necessarily reflect the position of the authors at the close of group discussion nor the final majority opinion of the group.