This application addresses broad Challenge Area (06) Enabling Technologies, 06-HL-109: Generate reagents for studying lung cell biology and disease progression. Chronic airway disease, including COPD, idiopathic fibrosis, bronchitis, asthma, and cystic fibrosis, affects millions of Americans. These diseases involve multiple cell types and undergo complex remodeling over time. There is a tremendous need of new markers and mouse models of these diseases to develop mitigating therapies. This is a proposal to generate both cell-specific and disease-specific monoclonal antibodies using broad immunization strategies combined with targeted screening approaches. In our preliminary, proof-of-concept experiments, we have generated more than 30 monoclonal antibodies specific to ciliated cells of the trachea. Some of these monoclonal antibodies reveal subpopulations of ciliated cells not apparent from histology alone. We now want to greatly expand these efforts to target a broad spectrum of cells from both the upper and lower airways. In addition, we want to identify disease-specific markers by immunizing mice with airway tissues derived from rat models of allergic asthma, COPD, and other chronic conditions. We will then perform molecular characterizations of the target proteins of these monoclonal antibodies, and then introduce GFP into the genomic sequences of their cognate genes to test for cell-specific expression. Finally, we will generate libraries of monoclonal antibodies specific to cell surface proteins of airway epithelia for tools in FACS analysis of airway disease. We anticipate that these libraries of monoclonal antibodies and tissue-specific promoters will become vital tools for the genetic and molecular analysis of airway disease analysis and progression.
20 million Americans suffer from some form of chronic airway disease. This is a proposal to identify new molecular tools to evaluate the complex tissue changes that accompany these diseases to empower research to understand these changes and design means of reversing them.
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