Airway epithelial cell gene regulation: new mechanisms and therapeutic strategies

项目摘要 排列在气道上的上皮细胞包括多种特化的细胞类型，这些细胞在肺中起重要作用。 健康上皮细胞的变化是哮喘和其他常见肺部疾病的显著特征。期间 正常分化，基因表达的巨大变化是独特的细胞发育所必需的。 上皮内基底细胞、分泌细胞和纤毛细胞群的功能能力。更以 在呼吸道疾病中，每种细胞类型响应于致病性细胞因子具有独特的基因表达变化。 在过去的二十年里，我们已经使用转基因小鼠建模，人类细胞培养和临床研究 确定上皮细胞基因表达变化的分子机制， 阻塞性哮喘尽管我们和其他人已经从小鼠模型中获得了巨大的洞察力，但我们的总体研究结果仍然是： 了解哮喘和其他人类呼吸道疾病的生物学基础的目标越来越多地引导我们 专注于开发合适的人体实验系统。在过去的几年里，我们和我们的 合作者已经开发了一套强大的方法，包括单细胞RNA-seq，ChIP-seq，大规模 平行报告基因测定和CRISPR介导的基因编辑，以解决原发性肝癌的机制问题。 人气道上皮细胞这些方法现在使我们能够执行潜在的开创性 这些实验为气道上皮细胞的复杂性提供了新的见解， 表达和上皮细胞功能，并揭示如何特定的气道上皮基因调节剂有助于 哮喘和其他呼吸道疾病。大多数与疾病相关的遗传变异都存在于调节区， 我们的工作对于理解基因对气道疾病风险的贡献具有重要意义。我们将 还致力于应用对基因调控的见解，以开发治疗性重编程的方法。 上皮以预防、治愈或治疗疾病。加州大学旧金山分校的环境提供了创造性的合作者， 尖端技术和关键的临床资源，并提供出色的机会， 职业调查员，他们将有能力继续推动实地工作向前发展。

PROJECT SUMMARY The epithelium that lines the airway comprises a variety of specialized cell types that play essential roles in lung health. Changes in the epithelium are prominent features of asthma and other common lung diseases. During normal differentiation, dramatic changes in gene expression are required for the development of the unique functional capabilities of the basal, secretory, and ciliated cell populations within the epithelium. Furthermore, in airway disease, each cell type has distinctive gene expression changes in response to pathogenic cytokines. Over the past two decades, we have used transgenic mouse modeling, human cell culture, and clinical studies to identify molecular mechanisms that underlie changes in epithelial gene expression that contribute to airway obstruction in asthma. Although we and others have gained tremendous insights from mouse models, our overall goal of understanding the biology underlying asthma and other human airway diseases has increasingly led us to focus on developing appropriate human experimental systems. In the past several years, we and our collaborators have developed a set of powerful approaches including single cell RNA-seq, ChIP-seq, massively parallel reporter assays, and CRISPR-mediated gene editing, to address mechanistic questions in primary human airway epithelial cells. These approaches now enable us to perform potentially groundbreaking experiments that give new insights into airway epithelial cell complexity, identify key regulators of gene expression and epithelial cell function, and reveal how specific airway epithelial gene regulators contribute to asthma and other airway diseases. Most disease-associated genetic variants are found in regulatory regions, and our work has important implications for understanding genetic contributions to airway disease risk. We will also work to apply insights about gene regulation to develop approaches for therapeutic reprogramming of the epithelium to prevent, cure, or treat disease. The environment at UCSF provides access to creative collaborators, cutting-edge technologies, and key clinical resources and affords outstanding opportunities for working with early career investigators who will be well positioned to continue to move the field forward.