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：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COPD、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。这些疾病的新标记物和小鼠模型的巨大需求，以开发缓解疗法。这是一个建议，以产生细胞特异性和疾病特异性的单克隆抗体，使用广泛的免疫策略结合靶向筛选方法。在我们初步的概念验证实验中，我们已经产生了30多种特异性针对气管纤毛细胞的单克隆抗体。其中一些单克隆抗体揭示了纤毛细胞的亚群，仅从组织学上看不明显。我们现在希望大大扩展这些努力，以针对上呼吸道和下呼吸道的广谱细胞。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气道组织免疫小鼠来鉴定疾病特异性标志物。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，我们将产生特异性针对气道上皮细胞表面蛋白的单克隆抗体库，用于气道疾病的流式细胞仪分析。我们预期这些单克隆抗体和组织特异性启动子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。 公共卫生相关性：2000万美国人患有某种形式的慢性气道疾病。这是一项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的建议，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.