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: 8041456
• 负责人: JOHN F ENGELHARDT
• 金额: $ 42.1万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-05-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8041456
• 关键词: Ablation; Address; Air; Anti-Bacterial Agents; Asthma; Binding; Binding Sites; Biological Assay; Biological Models; Biology; Cell Line; Cell physiology; Cells; Characteristics; Chromatin; Chronic Bronchitis; Collagen; Complex; Cystic Fibrosis; Development; Disease; Dissection; Duct (organ) structure; Ectopic Expression; Elements; Enhancers; Epithelial; Epithelial Cells; Epithelium; Ferrets; Funding; Ganciclovir; Gel; Gene Expression; Genetic; Genetic Models; Gland; Goals; Grant; Growth; HMG Domain; HMG-Box; Human; Hyperplasia; Hypertrophy; In Vitro; Indium; Infection; Injury; Interphase; Intrinsic factor; Knock-in Mouse; Knockout Mice; Knowledge; Label; Laboratories; LacZ Genes; Link; Liquid substance; Lung; Lung diseases; Lymphoid; Maintenance; Mediating; Mesenchymal; Methods; Modeling; Molecular; Morphogenesis; Mucous body substance; Mus; Mutation; Natural Immunity; Nose; Pathologic; Pathway interactions; Pattern; Phenotype; Physiology; Play; Production; Regulation; Regulatory Pathway; Reporter; Repression; Role; Serous; Signal Transduction; Signaling Molecule; Small Interfering RNA; Staging; Stem cells; Surface; System; TCF7L2 gene; Testing; Time; Tracheal Epithelium; Transcriptional Activation; Transcriptional Regulation; Transgenic Animals; Transgenic Mice; Transgenic Organisms; Viral Vector; Viscosity; Wnt proteins; Xenograft procedure; airway epithelium; animal model development; base; bone morphogenetic protein 2; cell type; enhancing factor; gland development; in vivo; mouse model; novel therapeutic intervention; progenitor; promoter; reconstitution; stem cell biology; stem cell fate; stem cell niche; time use; tool; transcription factor; Ablation; Address; Air; Anti-Bacterial Agents; Asthma; Binding; Binding Sites; Biological Assay; Biological Models; Biology; Cell Line; Cell physiology; Cells; Characteristics; Chromatin; Chronic Bronchitis; Collagen; Complex; Cystic Fibrosis; Development; Disease; Dissection; Duct (organ) structure; Ectopic Expression; Elements; Enhancers; Epithelial; Epithelial Cells; Epithelium; Ferrets; Funding; Ganciclovir; Gel; Gene Expression; Genetic; Genetic Models; Gland; Goals; Grant; Growth; HMG Domain; HMG-Box; Human; Hyperplasia; Hypertrophy; In Vitro; Indium; Infection; Injury; Interphase; Intrinsic factor; Knock-in Mouse; Knockout Mice; Knowledge; Label; Laboratories; LacZ Genes; Link; Liquid substance; Lung; Lung diseases; Lymphoid; Maintenance; Mediating; Mesenchymal; Methods; Modeling; Molecular; Morphogenesis; Mucous body substance; Mus; Mutation; Natural Immunity; Nose; Pathologic; Pathway interactions; Pattern; Phenotype; Physiology; Play; Production; Regulation; Regulatory Pathway; Reporter; Repression; Role; Serous; Signal Transduction; Signaling Molecule; Small Interfering RNA; Staging; Stem cells; Surface; System; TCF7L2 gene; Testing; Time; Tracheal Epithelium; Transcriptional Activation; Transcriptional Regulation; Transgenic Animals; Transgenic Mice; Transgenic Organisms; Viral Vector; Viscosity; Wnt proteins; Xenograft procedure; airway epithelium; animal model development; base; bone morphogenetic protein 2; cell type; enhancing factor; gland development; in vivo; mouse model; novel therapeutic intervention; progenitor; promoter; reconstitution; stem cell biology; stem cell fate; stem cell niche; time use; tool; transcription factor

Submucosal glands in the cartilaginous airways are thought to play important roles in protecting the human lung from infection by secreting antibacterial factors and controlling the composition and viscosity of fluid in the airways. In diseases such as cystic fibrosis (CF), asthma, and chronic bronchitis, Submucosal glands expand in mass (hypertrophy) and/or abundance (hyperplasia), leading to excessive and abnormal mucus production. Genetic defects in serous cells of submucosal glands have also been hypothesized to be a contributing factor in lung disease in CF. In addition to playing a role in innate immunity of the airways, submucosal gland ducts have also recently been recognized as a potential protective niche for proximal airway stem cells. During the previous two funding cycles of this grant, our laboratory has focused on elucidating the mechanisms of submucosal gland development from proximal airway progenitors using human, ferret, and mouse model systems. The goal of these studies has been to better characterize the phenotype of glandular progenitor cells by understanding the transcriptional pathways that control the growth of glands in the airway during normal development. Since submucosal gland progenitors also appear to have the capacity to contribute to surface airway epithelial cell renewal, the biology of stem cells in both submucosal gland and surface epithelial compartments may be closely linked. We have previously demonstrated that a transcription factor, lymphoid enhancing factor-1 (Lef-1), is induced during the initial stages of airway progenitor/stem cell commitment to form submucosal glands and is absolutely required for glands to develop in the airway. More recently, we have begun to dissect the regulatory pathways that control transcriptional activation of the Lef-1 promoter in cell line models and glandular progenitors in transgenic mice. We have found that Wnt/(3-catenin pathways appear to play a critical role in regulating the Lef-1 promoter through a set of related HMG-box transcription factors (TCP and Sox). This proposal will use in vitro polarized airway models, exvivo xenograft airway models, and genetic mouse models to study how Wnts, Noggin, and BMPs coordinate TCF/Sox regulation of the Lef-1 promoter during submucosal gland morphogenesis. Based on the inhibitory and enhancer functions of an identified Wnt-responsive element in the Lef-1 promoter, we hypothesize that Lef-1 regulation in glandular progenitors is controlled by both inhibitory (Sox and BMPs) and inductive (Wnt, TCF, and Noggin) signals. These hypotheses will be tested using viral vector systems and knockout mouse models capable of modulating the abundance of both extrinsic secreted factors and intrinsic transcription factors important for maintaining progenitor/stem cell phenotype and/or commitment to form glands. Lastly, this proposal will also attempt to evaluate whether TCF/Lef-1 pathways play a role in maintaining and/or mobilizing glandular duct stem cell niches for surface airway epithelial renewal following injury. Ultimately, this project will increase our understanding of stem cell phenotypes in the airway that have multipotent capacity for submucosal gland development and potentially also surface airway epithelial renewal. An increased understanding of submucosal gland morphogenesis may aid in the development of new therapeutic approaches to treat submucosal gland hyperplasia and hypertrophy in hypersecretory lung diseases. Furthermore, a greater knowledge of stem cell biology in the airway will greatly benefit the development genetic-based therapies for inhibited disease such as CF.

据认为，软骨气道中的粘膜粘膜在保护人肺免受保护方面起着重要作用 通过分泌抗菌因子并控制气道中流体的组成和粘度来感染。在 诸如囊性纤维化（CF），哮喘和慢性支气管炎等疾病，粘膜下腺在质量（肥大）中膨胀 和/或丰度（增生），导致过度和异常的粘液产生。浆液性细胞中的遗传缺陷 粘膜下腺也被认为是CF中肺部疾病的一个促成因素。除了玩 粘膜粘膜导管在天生的免疫力中的作用，最近也被认为是潜在的 近端气道干细胞的保护性利基市场。在这笔赠款的前两个资金周期中，我们的实验室有 专注于阐明使用近端气道祖细胞发育的粘膜粘膜发育机制 人，雪貂和小鼠模型系统。这些研究的目的是更好地表征 腺祖细胞通过了解控制气道腺体生长的转录途径 在正常发展期间。由于粘膜粘膜祖细胞似乎也有能力有助于表面 气道上皮细胞更新，粘膜腺和表面上皮室中干细胞的生物学 可能是紧密联系的。我们以前已经证明了转录因子淋巴增强因子1（LEF-1）， 在气道祖细胞/干细胞的初始阶段诱导，以形成粘膜粘膜腺体，IS 腺体在气道中发育绝对必需。最近，我们开始剖析调节途径 在细胞系模型和转基因中，LEF-1启动子的转录转录激活 老鼠。我们发现Wnt/（3-catenin途径似乎在通过A调节LEF-1启动子方面起着关键作用 一组相关的HMG-box转录因子（TCP和SOX）。该提案将使用体外偏光气道模型，Exvivo 异种移植气道模型和遗传小鼠模型，以研究WNT，Noggin和BMP如何坐标TCF/SOX 在粘膜下腺形态发生过程中，LEF-1启动子的调节。基于抑制和增强子功能 LEF-1启动子中鉴定出的WNT响应元件的识别，我们假设LEF-1在腺体中调节 祖细胞受抑制（SOX和BMP）和电感（Wnt，TCF和Noggin）信号的控制。这些 假设将使用病毒矢量系统和敲除鼠标模型进行测试 外部分泌因子和固有转录因子对于维持祖细胞/干细胞表型很重要 和/或承诺形成腺体。最后，该建议还将尝试评估TCF/LEF-1途径是否发挥作用 在维持和/或动员腺管干细胞壁细胞壁细胞壁细胞壁细胞中的作用，以进行表面气道上皮更新 受伤。最终，该项目将增加我们对具有多功能的气道中的干细胞表型的理解 粘膜粘膜发育的能力以及可能表面呼吸道上皮更新。增加 了解粘膜粘膜腺形态发生可能有助于开发新的治疗方法 粘膜下腺增生和肥大性肺部疾病。此外，对 气道中的干细胞生物学将极大地使发展基于基因的遗传疗法用于抑制疾病，例如 参见