Monoclonal Antibodies and Genetic Elements for Airway Disease

描述（由申请人提供）：该申请涉及广泛的挑战领域（06）使能技术，06- hl -109：生成用于研究肺细胞生物学和疾病进展的试剂。慢性气道疾病，包括慢性阻塞性肺病、特发性纤维化、支气管炎、哮喘和囊性纤维化，影响着数百万美国人。这些疾病涉及多种细胞类型，并随着时间的推移经历复杂的重塑。对于这些疾病的新标记物和小鼠模型，开发缓解治疗方法的需求非常大。这是一项利用广泛免疫策略结合靶向筛选方法产生细胞特异性和疾病特异性单克隆抗体的建议。在我们初步的概念验证实验中，我们已经产生了30多种针对气管纤毛细胞的单克隆抗体。其中一些单克隆抗体显示纤毛细胞亚群，仅从组织学上看不明显。我们现在想要极大地扩展这些努力，以针对来自上呼吸道和下呼吸道的广泛细胞。此外，我们希望通过使用来自过敏性哮喘、COPD和其他慢性疾病大鼠模型的气道组织免疫小鼠来识别疾病特异性标记物。然后，我们将对这些单克隆抗体的靶蛋白进行分子表征，然后将GFP引入其同源基因的基因组序列中，以测试细胞特异性表达。最后，我们将建立针对气道上皮细胞表面蛋白的单克隆抗体文库，用于气道疾病的FACS分析工具。我们预计这些单克隆抗体和组织特异性启动子文库将成为气道疾病分析和进展的遗传和分子分析的重要工具。

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE: 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.