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.

项目摘要 众所周知，气道粘膜粘膜粘膜（SMG）含有表面气道上皮的储备干细胞 （SAE）。在老鼠中，这种利基仅服务于气管。但是，在人类，猪和雪貂等较大的哺乳动物中， SMG存在于整个叶内软骨气道中，可能会提供更广泛的功能 在疾病环境中维持近端导电通道上皮。在前六个资金中 周期，该赠款使用了小鼠和雪貂遗传模型来解决气道SMG生物学的多个方面， SMG茎/祖细胞生物学和囊性纤维化（CF）肺发病机理。该建议旨在确定 参与气道修复的腺体肌上皮细胞（GMEC）以及Wnt-的亚群 调节受伤后控制其行为的机制。根据我们的初步数据，我们假设 LEF-1和SOX9转录因子差异地控制Wnt响应的GMEC指出，该系数协调了 GMEC在气道表面上的承诺，更新，迁移和增殖。 AIM 1将定义 小鼠气管GMEC的生物学，并利用一系列转基因线（交叉谱系跟踪， 有条件的敲除，Wnt-Reporters，DOX诱导的H2B-GFP）研究LEF-1和SOX9参与 调节控制GMEC承诺，更新，迁移和增殖的过程。 AIM 2将使用 在雪貂中进行截面谱系跟踪，以定义GMEC亚型和腺体导管细胞的参与 保持体内稳态和受伤后保持外面的sae和叶内SAE。我们假设腺体 管道是GMEC成熟到基本细胞状态的一个地点，并且这种基本的分层关系 在慢性气道疾病（例如CF）中受到干扰。 AIM 3将通过识别障碍来检验这一假设 使用VX-770响应性，在轻度和重度CF肺部疾病的情况下，GMEC和腺体管壁细分市场 CFTRG551D雪貂。这些研究的新方面包括物种中的第一个非稳定命运映射（雪貂） 密切对人类的CF肺病和SMG生物学进行了密切建模，并支持命运图数据 证明雪貂GMEC（ACTA2CREER）和腺体导管细胞（KRT7CREER）参与SAE修复。这 研究还将阐明SMG干细胞室室内的行为差异和 叶内的软骨气道，这在小鼠中无法解决，因为它们缺乏aLobar中的SMG 航空公司。该项目旨在增强我们对气道SMG和 调节其参与SMG和表面气道修复的机制。鉴于Gmecs可以 再生腺体和表面气道细胞类型，它们是CF和 从该提案中获得的知识将增强这种努力。此外，这项工作将描绘 疾病相关的SMG干细胞生态位变化可能对CF气道的发病机理很重要 疾病和其他影响SMG的疾病，例如哮喘和慢性支气管炎。