Identification of a Wnt/ Beta-catenin responsive adult lung epithelial progenitor cell for tissue repair in chronic lung disease

项目总结/摘要 这项提议的目的是确定内源性上皮祖细胞的潜力， 肺气肿治疗的再生方法。肺气肿是慢性阻塞性肺疾病的主要表型， 肺疾病（COPD），影响美国超过400万人。它的特点是逐渐丧失 肺泡肺组织没有治疗，停止或扭转疾病。重要的是， 远端肺激活自我修复机制是有缺陷的肺气肿，提出了一个问题，为什么和 再生途径和/或细胞在肺气肿中被沉默。WNT/β-连环蛋白信号传导最近被 被确定为一种潜在的再生途径，在肺气肿中活性降低，WNT/β- 已显示连环蛋白信号传导启动内在肺组织修复。然而，这些机制和 响应WNT/β-连环蛋白信号传导并有助于组织修复的潜在祖细胞的鉴定 慢性肺部疾病的破坏，仍然未知。我使用转基因Wnt/β-catenin的初步研究 活性报告小鼠和祖细胞衍生的3D肺类器官测定已经鉴定出独特的肺上皮细胞， 形成气道和肺泡类器官并对以下调节表现出高敏感性的群体： WNT/β-连环蛋白信号传导，因此代表WNT/β-连环蛋白应答性祖细胞群体。此外，本发明还 群体和单细胞转录组分析已经导致了气道俱乐部细胞祖细胞的鉴定 在WNT/β-连环蛋白应答性群体中，作为主要细胞类型的细胞群。因此，本提案的目的是 是为了解决WNT/β-连环蛋白信号激活俱乐部细胞再生肺泡的中心假设， 肺气肿的肺组织该假设将在3个特定目标中进行检验：1）。测试的假设 俱乐部细胞群是WNT/β-连环蛋白应答性的并形成类器官; 2）。决定命运， 小鼠肺气肿模型中俱乐部细胞群的体内表型和它们的再生能力 WNT/β-连环蛋白激活; 3）.检验俱乐部细胞群对WNT/β-连环蛋白应答的假设 在人肺气肿肺组织中的激活。 拟议的工作将通过定义精确的WNT/β-连环蛋白反应俱乐部细胞来解决现有的挑战 在小鼠和人肺中的群体，并通过鉴定新的标记基因，允许阐明它们的 改变和再生潜力。这项工作和综合培训计划将使 研究者建立关于肺损伤和再生的全面知识， 随后确定导致未来成功肺组织再生的分子机制 独立的研究生涯。

Project Summary/Abstract The goal of this proposal is to determine the potential of endogenous epithelial progenitor cells to contribute to regenerative approaches for emphysema therapies. Emphysema is a major phenotype of chronic obstructive lung disease (COPD) and affects over 4 million people in the US. It is characterized by progressive loss of alveolar lung tissue without therapies that stop or reverse the disease. Importantly, the endogenous ability of the distal lung to activate self-repair mechanisms is defective in emphysema, raising the question why and which regenerative pathways and/or cells are silenced in emphysema. WNT/beta-catenin signaling has been recently identified as a potential regenerative pathway with reduced activity in emphysema and re-activation of WNT/beta- catenin signaling has been shown to initiate intrinsic lung tissue repair. However, the mechanisms and the identity of potential progenitor cells that respond to WNT/β-catenin signaling and contribute to the repair of tissue destruction in chronic lung diseases, remain unknown. My preliminary studies using transgenic Wnt/beta-catenin activity reporter mice and a progenitor cell derived 3D lung organoid assay have identified a unique lung epithelial population which forms airway and alveolar organoids and exhibits high sensitivity to the modulations of WNT/beta-catenin signaling, thus representing a WNT/beta-catenin responsive progenitor population. Further, population and single cell transcriptome analysis has led to the identification of an airway club cell progenitor population as the major cell type in the WNT/beta-catenin responsive population. Thus, the aim of this proposal is to address the central hypothesis that WNT/beta-catenin signaling activates club cells to regenerate alveolar tissue in the emphysematous lung. This hypothesis will be tested in 3 Specific Aims: 1). Test the hypothesis that the club cell population is WNT/beta-catenin responsive and forms organoids; 2). Determine the fate and phenotype of the club cell population in murine emphysema models in vivo and their regenerative capacity upon WNT/beta-catenin activation; 3). Test the hypothesis that the club cell population respond to WNT/beta-catenin activation in human emphysematous lung tissue. The proposed work will solve an existing challenge by defining a precise WNT/beta-catenin-responsive club cell population in both mouse and human lungs and by identifying novel marker genes allowing to elucidate their alterations and regenerative potential in emphysema. This work and the integrated training plan will allow the investigator to establish comprehensive knowledge on lung injury and regeneration and expertise and skills to subsequently define the molecular mechanism leading to successful lung tissue regeneration in a future independent research career.