PATHOBIOLOGY OF BRONCHIAL TISSUE KALLIKREIN

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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.