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.

概括 大多数呼吸系统疾病被认为是由于以下原因导致的异常或缺乏有效的修复机制： 受伤。识别可增强内源再生的细胞来源和机制 这可以帮助基于细胞的疗法是非常需要的。在我们最近发表的工作中，我们发现了一种新颖的储备 气道组织中的多能干细胞群。我们发现肌上皮细胞存在于 气道粘膜下腺 (SMG) 通常非常静止，但它们会广泛增殖并迁移 在严重损伤后重新填充气道表面上皮（SAE）。我们进一步鉴定了 SOX9 介导的 转录程序对于 SMG 衍生的肌上皮细胞增殖和迁移至 SAE。此外，我们发现一小部分 SMG 衍生细胞具有完全转化为 SAE 的能力，尽管 非常慢。我们目前的初步数据表明，SMG 衍生细胞中未转化为 SAE细胞继续保持SMG细胞特征，包括转录因子SOX9的表达， 对于较长的时间段。此外，使用新开发的小鼠模型和活体成像研究我们的 初步数据表明SMG-腺泡管腔细胞（浆液性和粘液性细胞）也具有迁移能力 到 SAE。根据我们的初步数据，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 还假设 SOX9 依赖性机制必须下调才能完整 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 表面上皮并增强适当的再生。在目标1中，我们将定性和定量 确定 SMG-腺泡管腔细胞对损伤后 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与多光子辅助活体成像相结合，以确定 SMG-腺泡管腔细胞迁移的能力及其对 SAE 修复的贡献。我们还会使用白喉毒素 介导的 MEC 消融来检验我们的假设，即 SMG 腺泡腔细胞增殖和迁移是 依赖 MEC。在 Aim2 中，我们将检验我们的假设，即 SOX9 的丢失足以完成完整的任务。 在急性损伤和慢性炎症的情况下，SMG 衍生细胞转化为 SAE 细胞。这部作品 变得更加重要，因为我们发现 SMG 衍生的细胞对 SAE 再生的贡献发生在 多种形式的伤害环境，包括流感病毒、二氧化硫、氯和慢性过敏原 诱发气道损伤。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。这项工作将为今后的学习奠定基础 涉及人类气道再生。