Impact of alcohol on lung fibroblast regulation of the alveolar epithelial barrier

酒精对肺成纤维细胞调节肺泡上皮屏障的影响

• 批准号: 9896468
• 负责人: VIRANUJ SUEBLINVONG
• 金额: $ 7.8万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-15 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9896468
• 关键词: Acute; Adult Respiratory Distress Syndrome; Alcohols; Alveolar; Alveolar Macrophages; Animals; Attenuated; Automobile Driving; Bleomycin; Cell Differentiation process; Cell physiology; Chronic; Chronic Obstructive Airway Disease; Chronic lung disease; Coculture Techniques; Collagen; Communication; Complex; Cytokine Signaling; Data; Deposition; Down-Regulation; Endotoxins; Epithelial; Epithelial Cells; Epithelium; Ethanol; Experimental Models; Extracellular Matrix; Extracellular Matrix Proteins; Failure; Fibroblasts; Fibronectins; Foundations; Functional disorder; Future; Goals; Growth Factor; Homeostasis; Impairment; Individual; Injury; Interstitial Lung Diseases; Laboratories; Lead; Lung; Lung diseases; Measures; Mesenchymal; MicroRNAs; Modality; Molecular; Morbidity - disease rate; Normal tissue morphology; Organ; Oxidative Stress; Pathologic; Phagocytosis; Phase; Populations at Risk; Preventive Intervention; Process; Production; Proteins; Quality of life; Recovery; Regulation; Resistance; Resolution; Risk; Role; Signal Pathway; Signal Transduction; Signaling Molecule; Survivors; Testing; Therapeutic; Therapeutic Intervention; Tight Junctions; Tissues; Transforming Growth Factor beta; Up-Regulation; alcohol effect; alcohol exposure; alveolar epithelium; cell injury; cell motility; cell type; chronic alcohol ingestion; cytokine; design; equilibration disorder; exosome; extracellular vesicles; insight; intercellular communication; lung injury; lung repair; mortality; novel; prevent; problem drinker; repaired; response; restoration; tissue injury; tissue repair; wound

Project Summary Tissue disrepair following injury leads to organ dysfunction and increases morbidity and mortality of those who survive the initial insult. In the lung, the disrepair process is associated with excessive collagen deposition along with failure of both re-epithelialization and epithelial cell tight junction formation. Tissue disrepair is associated with dysregulation of TGFβ signaling in a variety of pulmonary diseases including the acute respiratory distress syndrome (ARDS), chronic obstructive pulmonary disease (COPD), and interstitial lung disease (ILD). Our laboratory utilizes experimental models of chronic alcohol ingestion and bleomycin- induced lung injury to assess the molecular mechanisms of lung repair and identify novel preventative and therapeutic interventions. We have previously shown that alcohol induces excessive and persistent TGFβ expression in the lung. Additionally, we demonstrated that in the lung of chronic alcohol-exposed animals, TGFβ is a critical molecule driving many cellular anomalies by increasing airway oxidative stress, decreasing alveolar macrophages phagocytosis, and priming the lung for fibroproliferative disrepair following acute injury. Our preliminary data show that alcohol-exposed lung fibroblasts interfere with epithelial cell barrier formation likely through induction of epithelial-mesenchymal transition (EMT). Additionally, inhibition of TGFβ signaling attenuates the effect of alcohol-exposed fibroblasts on epithelial cells. Interestingly, we also showed that fibroblasts influence epithelial cells indirectly via fibroblast-derived exosomes rather than direct secretion of cytokines or growth factors. Furthermore, we showed that alcohol disturbs the balance of pro- and anti-fibrotic microRNA (miR) expression. Specifically, alcohol increases pro-fibrotic miR-21 and attenuates anti-fibrotic miRNA-1946a in lung fibroblasts. These data lead us to hypothesize that alcohol exposure disrupts alveolar epithelial cell tight junction formation and barrier function following injury through an imbalance of miR-21 and miR-1946a in exosomes secreted by lung fibroblasts. The experimental approaches are designed to test this hypothesis, and these studies are expected to provide a firm scientific basis for the underlying mechanism(s) by which alcohol interferes with normal repair following lung injury. The results from this proposal will set the basis for future studies to investigate potential therapeutic strategies to prevent or mitigate tissue injury and disrepair in the at-risk population (i.e., alcoholic individuals).

项目摘要 损伤后的组织破损导致器官功能障碍，并增加了糖尿病的发病率和死亡率。 那些在最初的侮辱中幸存下来的人。在肺中，破损过程与过度的胶原蛋白有关 沉积沿着，同时上皮再形成和上皮细胞紧密连接形成失败。组织 在多种肺部疾病中，肺组织损伤与TGFβ信号转导失调有关， 急性呼吸窘迫综合征（ARDS）、慢性阻塞性肺疾病（COPD）和间质性 肺疾病（ILD）。我们的实验室利用慢性酒精摄入和博来霉素的实验模型- 诱导的肺损伤，以评估肺修复的分子机制，并确定新的预防和 治疗干预。我们先前已经表明，酒精诱导过度和持久的TGFβ 在肺部的表达。此外，我们证明了在慢性酒精暴露动物的肺中， TGFβ是通过增加气道氧化应激、减少气道炎症和/或呼吸道炎症而驱动许多细胞异常的关键分子。 肺泡巨噬细胞吞噬作用，并引发肺纤维增生性损伤后急性损伤。 我们的初步数据表明，暴露于酒精的肺成纤维细胞会干扰上皮细胞屏障的形成 可能通过诱导上皮-间质转化（EMT）。此外，抑制TGFβ信号传导 减弱暴露于酒精的成纤维细胞对上皮细胞的影响。有趣的是，我们还发现， 成纤维细胞通过成纤维细胞来源的外泌体间接影响上皮细胞，而不是直接分泌 细胞因子或生长因子。此外，我们发现酒精干扰了促纤维化和抗纤维化的平衡。 microRNA（miR）表达。具体而言，酒精增加促纤维化miR-21并减弱抗纤维化miR-21。 肺成纤维细胞中的miRNA-1946 a。这些数据使我们假设，酒精暴露破坏肺泡 上皮细胞紧密连接的形成和屏障功能损伤后通过miR-21和 肺成纤维细胞分泌的外泌体中的miR-1946 a。实验方法旨在验证这一点 这些研究有望为潜在机制提供坚实的科学依据 酒精会干扰肺损伤后的正常修复。该提案的结果将确定 未来研究的基础，以调查潜在的治疗策略，以防止或减轻组织损伤， 在危险人群中的破损（即，酗酒者）。