Monoclonal Antibodies and Genetic Elements for Airway Disease

描述(由申请人提供)：本申请涉及广泛的挑战领域(06)使能技术，06-HL-109：产生用于研究肺细胞生物学和疾病进展的试剂。慢性呼吸道疾病，包括慢性阻塞性肺疾病、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。迫切需要这些疾病的新标记物和小鼠模型来开发缓解疗法。这是一项建议，利用广泛的免疫策略和有针对性的筛查方法相结合，产生细胞特异性和疾病特异性的单抗。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抗。其中一些单抗显示纤毛细胞亚群，仅从组织学上看并不明显。我们现在希望极大地扩大这些努力，以针对来自上呼吸道和下呼吸道的广泛范围的细胞。此外，我们希望通过用来自过敏性哮喘、COPD和其他慢性疾病大鼠模型的呼吸道组织免疫小鼠来识别疾病特异性标志物。然后，我们将对这些单抗的目标蛋白进行分子表征，然后将GFP引入它们的同源基因的基因组序列中，以测试细胞特异性表达。最后，我们将建立针对呼吸道上皮细胞表面蛋白的单抗的文库，为FACS分析呼吸道疾病提供工具。我们预计，这些单抗和组织特异性启动子文库将成为分析和发展呼吸道疾病的遗传和分子分析的重要工具。 与公共卫生相关：2000万美国人患有某种形式的慢性呼吸道疾病。这是一项建议，旨在确定新的分子工具来评估伴随这些疾病的复杂组织变化，以使研究能够理解这些变化并设计逆转它们的方法。

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.