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

项目摘要/摘要 这项提议的目标是确定内源性上皮祖细胞对 肺气肿治疗的再生方法。肺气肿是慢性阻塞性疾病的主要表现型。 肺部疾病(COPD)，在美国有400多万人受到影响。它的特点是逐渐失去 未经治疗阻止或逆转疾病的肺泡组织。重要的是，人的内生能力 肺气肿患者的远端肺激活自我修复机制是有缺陷的，这提出了一个问题，为什么？ 在肺气肿中，再生途径和/或细胞被沉默。WNT/β-连环蛋白信号转导最近 被认为是一种潜在的再生途径，在肺气肿中活性降低，WNT/β-1重新激活- 连环蛋白信号已被证明启动肺组织的内在修复。然而，这些机制和 应答Wnt/β-连环蛋白信号并促进组织修复的潜在祖细胞的鉴定 慢性肺部疾病的破坏情况仍不得而知。转基因Wnt/β-连环蛋白的初步研究 活动报告小鼠和祖细胞衍生的3D肺器官分析已经确定了一种独特的肺上皮细胞 形成呼吸道和肺泡类有机物质的种群，对 WNT/β-连环蛋白信号转导，因此代表了WNT/β-连环蛋白应答的前体种群。此外， 群体和单细胞转录组分析导致了呼吸道俱乐部细胞前体细胞的鉴定 在WNT/β-连环蛋白应答群体中，群体是主要的细胞类型。因此，这项提案的目的是 是为了解决中心假设，即WNT/β-连环蛋白信号激活俱乐部细胞再生肺泡 肺气肿肺组织。这一假设将在三个具体目标上得到检验：1)。检验以下假设 俱乐部细胞群对WNT/β-连环蛋白有反应，并形成类器官；决定命运和 小鼠肺气肿模型中俱乐部细胞群的表型及其再生能力 WNT/β-连环蛋白激活；检验俱乐部细胞群对WNT/β-连环蛋白有反应的假设 人肺气肿肺组织中的活化。 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 通过识别新的标记基因来阐明它们在小鼠和人类肺中的分布 肺气肿的改变和再生潜力。这项工作和综合培训计划将使 调查员将建立关于肺损伤和再生的全面知识以及专业知识和技能 随后明确了导致未来成功肺组织再生的分子机制 独立研究生涯。

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.