Biology of Submucosal Gland Stem Cells in the Airway

气道粘膜下腺干细胞的生物学

• 批准号: 10649543
• 负责人: JOHN F ENGELHARDT
• 金额: $ 70.18万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-20 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10649543
• 关键词: Acute; Address; Adult; Affect; Airway Disease; Asthma; Basal Cell; Behavior; Biology; Caucasians; Cell Compartmentation; Cell Lineage; Cell Maintenance; Cell Maturation; Cell surface; Cells; Chronic; Chronic Bronchitis; Chronic Phase; Chronic lung disease; Cre driver; Cystic Fibrosis; Data; Developmental Biology; Disease; Disease Progression; Duct (organ) structure; Ductal Epithelial Cell; Epithelial Cells; Family suidae; Ferrets; Functional disorder; Funding; Genes; Genetic Models; Gland; Glandular Cell; Goals; Grant; Homeostasis; Human; Human Biology; Injury; Knowledge; Label; Life; Lobar; Lung; Maintenance; Mammals; Maps; Modeling; Mus; Myoepithelial cell; Natural regeneration; Outcome; Pathogenesis; Pathologic; Patient Care; Phase; Phenotype; Process; Proliferating; Property; Pulmonary Cystic Fibrosis; Recording of previous events; Reporter; Repression; Research; Reserve Stem Cell; Signal Transduction; Site; Stress; Submucosa; Surface; System; Testing; Trachea; Transgenic Organisms; Translating; VX-770; WNT Signaling Pathway; Work; airway epithelium; airway regeneration; airway repair; autosome; cartilaginous; cell type; chronic respiratory disease; clinical care; conditional knockout; cystic fibrosis airway; design; epithelial repair; human model; injured airway; insight; migration; mouse model; novel; pressure; progenitor; regenerative; repaired; stem cell biology; stem cell function; stem cell niche; stem cells; tool; transcription factor

Project Summary Airway submucosal glands (SMGs) are known to contain reserve stem cells for the surface airway epithelium (SAE). In mice, this niche serves only the trachea; however, in larger mammals such as humans, pigs and ferrets, SMGs are present throughout the intralobar cartilaginous airways and may serve the broader function of maintaining the proximal conducting airway epithelium in the setting of disease. During the previous six funding cycles, this grant has used mouse and ferret genetic models to address multiple aspects of airway SMG biology, SMG stem/progenitor cell biology, and cystic fibrosis (CF) lung pathogenesis. This proposal aims to identify the subpopulations of glandular myoepithelial cells (GMECs) that participate in airway repair, as well as the Wnt- regulated mechanisms that control their behavior following injury. Based on our preliminary data, we hypothesize that Lef-1 and Sox9 transcription factors differentially control Wnt-responsive GMEC states that orchestrate the commitment, renewal, migration, and proliferative expansion of GMECs on the airway surface. Aim 1 will define the biology of tracheal GMECs in mice and utilize an array of transgenic lines (intersectional lineage tracing, conditional knockout, Wnt-reporters, Dox-inducible H2B-GFP) to study the involvement of Lef-1 and Sox9 in regulating processes that control GMEC commitment, renewal, migration, and proliferation. Aim 2 will use intersectional lineage tracing in ferrets to define the participation of GMEC subtypes and gland ductal cells in maintaining the extralobar and intralobar SAE at homeostasis and following injury. We hypothesize that gland ducts are a site for GMEC maturation to a pre-basal cell state, and that this underlying hierarchical relationship is disturbed in chronic airway diseases such as CF. Aim 3 will test this hypothesis by identifying disturbances in GMEC and gland duct niches in the setting of mild and severe CF lung disease, using VX-770-responsive CFTRG551D ferrets. Novel aspects of these studies include the first non-rodent fate mapping in a species (ferret) that closely models CF lung disease and SMG biology of humans, and supporting fate mapping data demonstrating that ferret GMECs (ACTA2CreER) and gland ductal cells (KRT7CreER) participate in SAE repair. This research will also shed light on differences in the behavior of SMG stem cell compartments in the extralobar and intralobar cartilaginous airways, which cannot be addressed in mice because they lack SMGs in the intralobar airways. This project is designed to enhance our understanding of stem cell phenotypes in airway SMGs and the mechanisms that regulate their participation in SMG and surface airway repair. Given that GMECs can regenerate both glandular and surface airway cell types, they are an attractive target for gene editing in CF and such efforts will be enhanced by knowledge gained from this proposal. Furthermore, this work will delineate disease-associated changes to SMG stem cell niches that may be important for the pathogenesis of CF airway disease and other hypersecretory diseases that affect SMGs, such as asthma and chronic bronchitis.

项目概要 已知气道粘膜下腺 (SMG) 含有气道表面上皮的储备干细胞 （美国汽车工程师学会）。在小鼠中，这个生态位只服务于气管；然而，在人类、猪和雪貂等大型哺乳动物中， SMG 存在于整个叶内软骨气道中，并且可以发挥更广泛的功能 在疾病情况下维持近端传导气道上皮。在前六次融资期间 周期，这笔赠款使用小鼠和雪貂遗传模型来解决气道 SMG 生物学的多个方面， SMG 干/祖细胞生物学和囊性纤维化 (CF) 肺发病机制。该提案旨在确定 参与气道修复的腺肌上皮细胞 (GMEC) 亚群以及 Wnt- 控制他们受伤后行为的调节机制。根据我们的初步数据，我们假设 Lef-1 和 Sox9 转录因子差异性地控制 Wnt 响应的 GMEC 状态，从而协调 气道表面 GMEC 的承诺、更新、迁移和增殖扩张。目标 1 将定义 小鼠气管 GMEC 的生物学，并利用一系列转基因系（交叉谱系追踪、 条件敲除、Wnt 报告基因、Dox 诱导的 H2B-GFP）来研究 Lef-1 和 Sox9 在 监管控制 GMEC 承诺、更新、迁移和扩散的流程。目标2将使用 雪貂中的交叉谱系追踪以确定 GMEC 亚型和腺导管细胞的参与 维持叶外和叶内 SAE 的稳态以及受伤后的状态。我们假设腺体 导管是 GMEC 成熟到前基底细胞状态的场所，并且这种潜在的层次关系 慢性气道疾病（如 CF）会受到干扰。目标 3 将通过识别干扰来检验这一假设 使用 VX-770 响应在轻度和重度 CF 肺部疾病中的 GMEC 和腺管生态位 CFTRG551D 雪貂。这些研究的新颖之处包括第一个物种（雪貂）的非啮齿动物命运图谱 密切模拟人类 CF 肺病和 SMG 生物学，并支持命运图谱数据 证明雪貂 GMEC (ACTA2CreER) 和腺导管细胞 (KRT7CreER) 参与 SAE 修复。这 研究还将揭示叶外和叶外 SMG 干细胞区室行为的差异。 叶内软骨气道，无法在小鼠中解决，因为它们在叶内缺乏 SMG 航空公司。该项目旨在增强我们对气道 SMG 中干细胞表型的理解， 调节其参与 SMG 和表面气道修复的机制。鉴于 GMEC 可以 再生腺体和表面气道细胞类型，它们是 CF 和 中基因编辑的有吸引力的目标 从该提案中获得的知识将加强此类努力。此外，这项工作将描绘 SMG 干细胞生态位的疾病相关变化可能对 CF 气道的发病机制很重要 疾病和其他影响 SMG 的分泌过多疾病，例如哮喘和慢性支气管炎。