My career goal is to become an independent physician-scientist investigating basic mechanisms of airway epithelial biology contributing to chronic respiratory diseases. This application outlines a comprehensive training program including an intensive laboratory research experience, a set of courses and seminars, and a mentoring system. The program builds upon my experiences as a neuroscience Ph.D. student and a clinical pulmonary fellow. My research project is inspired by exciting new discoveries made in the lab of my mentor, Dr. David Erle. Excessive amounts of mucins, enormous glycoproteins that appear to be critical for the viscoelastic properties of mucus, contribute to morbidity and mortality in chronic airway diseases such as asthma, COPD, and cystic fibrosis. Previous trainees in the Erle lab have shown that AGR2, a novel member of the protein disulfide isomerase (PDI) family, plays a selective role in mucus production in vivo. Mice lacking AGR2 are completely unable to produce intestinal mucin and have reduced airway mucin production in a mouse model of asthma. I hypothesize that AGR2 and a closely related PDI, AGR3, interact directly with airway mucins in the endoplasmic reticulum and that these interactions are critical for mucin production. AGR proteins may also interact with other proteins as they transit through the ER of mucous cells and perhaps other cells in the lung. I have 3 specific aims:
Aim 1 will analyze AGR2, AGR3 and mucin expression patterns in airway epithelium from naove and allergen- challenged mice using immunohistochemistry.
Aim 2 will analyze the ability of AGR2 and AGR3 to bind to mucins and other potential binding partners in airway epithelial cells.
Aim 3 will analyze the function of AGR2 and AGR3 by knocking down AGR2 and AGR3 in cultured human bronchial epithelial cells. These experiments will enhance our understanding of the basic cellular mechanisms important for airway production of mucins and determine whether AGR2 and AGR3 are promising targets for the treatment of mucus hypersecretion. I will gain valuable hands-on experience in cell culture, animal models of airway disease, molecular biology, and biochemistry. The lab work will be complemented by coursework and state of the art seminars to keep me abreast of recent developments in relevant fields and help me develop skills in writing and oral presentation. A committee comprised of senior pulmonary scientists and an internationally known expert in ER biology will monitor my progress and advise me. This integrated training program will provide me with the experience needed to begin a career as a successful independent investigator.
This proposal will investigate the role of novel proteins in mucus production within the airway. These studies will aid our understanding of the cellular processes underlying excessive mucus production in diseases such as asthma, emphysema, and cystic fibrosis.