Airway epithelial cell gene regulation: new mechanisms and therapeutic strategies

项目总结 呼吸道上皮由多种特殊类型的细胞组成，这些细胞在肺中起着重要作用。 健康。上皮细胞的改变是哮喘和其他常见肺部疾病的显著特征。在.期间 正常分化、基因表达的剧烈变化是唯一发育所需的 上皮内的基细胞、分泌细胞和纤毛细胞群的功能。此外，在 在呼吸道疾病中，每种细胞类型都有不同的基因表达变化，以响应致病细胞因子。 在过去的二十年里，我们使用转基因小鼠模型、人类细胞培养和临床研究。 确定导致呼吸道上皮细胞基因表达变化的分子机制 哮喘中的梗阻。尽管我们和其他人从老鼠模型中获得了巨大的洞察力，但我们的总体 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 合作者们已经开发出一套强大的方法，包括单细胞rna-seq、芯片-seq、大规模 平行的记者分析和CRISPR介导的基因编辑，以解决小学机械性问题 人呼吸道上皮细胞。这些方法现在使我们能够执行潜在的开创性工作 对呼吸道上皮细胞复杂性有新见解的实验，确定了基因的关键调控因子 表达和上皮细胞功能，并揭示特定的呼吸道上皮基因调节因子如何在 哮喘和其他呼吸道疾病。大多数与疾病相关的基因变异都是在调控区域中发现的， 我们的工作对理解基因对呼吸道疾病风险的贡献具有重要意义。我们会 也致力于应用对基因调控的见解来开发治疗重新编程的方法 用于预防、治愈或治疗疾病的上皮。加州大学旧金山分校的环境为创造性的合作者提供了通道， 尖端技术和关键临床资源，为早期合作提供了绝佳的机会 职业调查员将处于有利地位，继续推动该领域的发展。

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.