Mechanisms of submucosal gland cell mediated airway regeneration

粘膜下腺细胞介导气道再生的机制

• 批准号: 10444912
• 负责人: Purushothama Rao Tata
• 金额: $ 40.25万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-02 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10444912
• 关键词: Ablation; Acinus organ component; Acute; Address; Airway Disease; Allergens; Basal Cell; Biological Assay; Cell Proliferation; Cell Therapy; Cell Transplantation; Cells; Cessation of life; Characteristics; Chlorine; Chronic; Coupled; Data; Diphtheria Toxin; Emigrations; Epithelial; Epithelial Cells; Failure; Family suidae; Foundations; Future; Genetic Transcription; Gland; Goals; Human; Imaging Device; Inflammation; Inflammatory; Inflammatory Response; Inhalation; Injury; Knowledge; Lobular; Lung diseases; MUC5B gene; Mediating; Modeling; Molecular; Monitor; Movement; Mucous body substance; Multipotent Stem Cells; Myoepithelial cell; Naphthalene; Natural regeneration; Outcome; Population; Process; Proliferating; Publishing; Respiratory Disease; Respiratory Mucosa; Secretory Cell; Serous; Source; Structure of parenchyma of lung; Sulfur Dioxide; Surface; Testing; Therapeutic; Time; Tissues; Toxin; United States; Work; airway epithelium; airway regeneration; base; cell motility; cell type; clinically relevant; epithelial repair; epithelium regeneration; genetic manipulation; imaging study; imaging system; in vivo; influenzavirus; injured airway; intravital imaging; loss of function; lung injury; lung regeneration; migration; mouse model; multi-photon; novel; pathogen; prevent; programs; repaired; respiratory; response; single-cell RNA sequencing; stem cell population; transcription factor; transdifferentiation

SUMMARY Most respiratory diseases are thought to result from an aberrant or a lack of efficient repair mechanisms following injuries. Identifying the cellular sources and the mechanisms that can enhance the endogenous regeneration and that can aid cell-based therapies is much needed. In our recent published work, we identified a novel reserve multipotent stem cell population in the airway tissues. We found that myoepithelial cells that reside in the submucosal glands (SMG) of the airways are normally very quiescent but they proliferate extensively and migrate to repopulate surface airway epithelium (SAE) following severe damage. We further identified SOX9-mediated transcriptional programs are necessary for the proliferation and migration of SMG-derived myoepithelial cells to SAE. Furthermore, we found that a fraction of SMG-derived cells have the ability to fully convert into SAE, albeit very slowly. Our current preliminary data indicated that the fraction of SMG-derived cells that do not convert into SAE cells continue to maintain SMG cell characteristics, including the expression of transcription factor SOX9, for extended time periods. In addition, using newly developed mouse models and intra-vital imaging studies our preliminary data indicate that SMG-acinar luminal cells (serous and mucous cells) also have the ability to migrate to SAE. Based on our preliminary data, we hypothesize that SMG-acinar luminal cells have the ability to migrate and contribute to SAE regeneration and that this process is dependent on MECs migration. We also hypothesize that SOX9-dependent mechanisms must be downregulated for the complete conversion of SMG-derived cells into SAE cells. The major objectives of this proposal are to address both the potential contribution of SMG- acinar luminal cells to surface epithelium and to enhance proper regeneration. In Aim1, we will qualitatively and quantitatively determine the contribution of SMG-acinar luminal cells to SAE repair after injury. We will use our newly developed in vivo lineage tracing mouse models coupled with multi-photon assisted intravital imaging to determine the ability of SMG-acinar luminal cells migration and contribution to SAE repair. We will also use diphtheria toxin mediated ablation of MECs to test our hypothesis that SMG-acinar luminal cell proliferation and migration is dependent on MECs. In Aim2, we will test our hypothesis that loss of SOX9 is sufficient for the complete conversion of SMG-derived cells into SAE cells in the context of acute injury and chronic inflammation. This work has taken on added importance, as we found that SMG-derived cells contribution to SAE regeneration occurs in multiple forms of injury contexts including, influenza virus, Sulphur dioxide, chlorine, and chronic allergen- induced airway damage. Therefore, the outcomes from the proposed studies will have broader significance to airway diseases that occur due to defective regeneration. This work will lay the foundation for future studies involving human airway regeneration.

总结 大多数呼吸系统疾病被认为是由于以下异常或缺乏有效的修复机制造成的 受伤确定可以增强内源性再生的细胞来源和机制 并且可以帮助基于细胞的治疗是非常需要的。在我们最近发表的研究中，我们发现了一种新的保护区， 多能干细胞群在气道组织中。我们发现，肌上皮细胞，居住在 气道的粘膜下腺（SMG）通常非常静止，但它们广泛增殖并迁移 以在严重损伤后重新填充表面气道上皮（SAE）。我们进一步鉴定了SOX 9介导的 转录程序对于SMG衍生的肌上皮细胞的增殖和迁移是必需的， SAE。此外，我们发现一部分SMG衍生的细胞具有完全转化为SAE的能力， 慢慢地我们目前的初步数据表明，SMG衍生的细胞中不转化为 SAE细胞继续保持SMG细胞特征，包括转录因子SOX 9的表达， 在很长一段时间内。此外，使用新开发的小鼠模型和活体成像研究， 初步数据表明，SMG-腺泡腔细胞（浆液和粘液细胞）也具有迁移能力， 是SAE。基于我们的初步数据，我们假设SMG-腺泡腔细胞有能力 迁移并有助于SAE再生，并且该过程依赖于MEC迁移。我们 我还假设，SOX 9依赖性机制必须下调，以完成 SMG衍生细胞转化为SAE细胞。 该提案的主要目标是解决SMG-腺泡腔细胞的潜在贡献 以使上皮表面化并增强适当的再生。在目标1中，我们将定性和定量地 确定SMG-腺泡腔细胞对损伤后SAE修复的贡献。我们将使用我们新开发的 体内谱系追踪小鼠模型结合多光子辅助活体成像，以确定 SMG-腺泡腔细胞迁移的能力和对SAE修复的贡献。我们还会用白喉毒素 介导的MECs消融，以检验我们的假设，即SMG-腺泡腔细胞增殖和迁移是 依赖于MEC。在Aim 2中，我们将测试我们的假设，即SOX 9的缺失足以完成 在急性损伤和慢性炎症的情况下，SMG衍生的细胞转化为SAE细胞。这项工作 已经采取了额外的重要性，因为我们发现SMG衍生的细胞对SAE再生的贡献发生在 多种形式的损伤环境，包括流感病毒、二氧化硫、氯和慢性过敏原- 导致气道损伤。因此，拟议研究的结果将具有更广泛的意义， 由于再生缺陷而发生的气道疾病。本文的工作为以后的研究奠定了基础 涉及人体呼吸道再生。