Impact of alcohol exposure on unjamming the airway epithelium

酒精暴露对疏通气道上皮的影响

• 批准号: 10547794
• 负责人: Kristen Leigh Fowler
• 金额: $ 4.09万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10547794
• 关键词: Acute Respiratory Distress Syndrome; Adult; Affect; Air; Alcohols; Alveolus; Area; Attenuated; Biological Assay; Breast Cancer cell line; Bronchi; Cell Adhesion Molecules; Cell Nucleus; Cell Separation; Cell Shape; Cell model; Cell physiology; Cells; Cessation of life; Chronic; Complementary DNA; Complex; Defect; Dimerization; Disease; Disease model; Dyes; Electrical Resistance; Epithelial Cells; Epithelium; Ethanol; Exhibits; Focal Adhesion Kinase 1; Functional disorder; Gene Expression; Goals; Human; Image; Immobilization; Immune system; In Vitro; Infection; Injury; Integral Membrane Protein; Laboratory Finding; Lentivirus; Link; Liquid substance; Lung; Lung infections; MAPK3 gene; Measures; Microscopy; Modeling; Molecular; Morphology; Pathway interactions; Patients; Pattern; Persons; Phenotype; Phospho-Specific Antibodies; Phosphorylation; Physical shape; Predisposition; Proteins; Quantitative Reverse Transcriptase PCR; RNA; Rattus; Repression; Research; Risk; Role; Stretching; Syndrome; Testing; Tight Junctions; Time; Trachea; Transfection; United States; Western Blotting; airway epithelium; alcohol exposure; alcohol response; alcohol use disorder; alveolar epithelium; asthmatic patient; attenuation; cell motility; chronic alcohol ingestion; design; experimental study; improved; in vitro Model; inhibitor; junctional adhesion molecule; keratinocyte; kinase inhibitor; knock-down; lung health; migration; monolayer; non-alcoholic; pathogen; problem drinker; receptor; small hairpin RNA; small molecule; wound healing

PROJECT ABSTRACT Alcohol use disorder affects over 14 million adults and causes 88,000 deaths each year in the United States. Chronic alcohol use significantly increases people’s risk of developing lung infections and acute respiratory distress syndrome (ARDS). This increased sensitivity to injury is a condition known as alcoholic lung syndrome, and it is caused by dysfunction of the lung immune system and alveolar epithelial barrier. However, little is known about how alcohol impacts epithelial cells in the conducting airway, i.e. the trachea and bronchi, which are the first line of defense against infectious pathogens in the lungs. We have used primary airway epithelial cells isolated from healthy and alcoholic patients and differentiated them in vitro to examine the impact of alcohol on cell barrier function and morphology. Healthy differentiated primary airway epithelial cells exhibit a cobblestone- like pattern and become immobile once the monolayer has matured— a normal airway cell phenotype known as “jamming”. By contrast, airway epithelial cells isolated from chronic alcoholics remained migratory and in an “unjammed” state, since areas of stretched cells and swirls of cells appear in the monolayer. Additionally, rat tracheal cells grown and differentiated in vitro in the presence of ethanol remain unjammed while control monolayers become jammed. It is known that the transmembrane protein junctional adhesion molecule A (JAM- A) regulates epithelial cell migration, and it also regulates barrier function by controlling the paracellular flow of small molecules as part of tight junction complexes. Recently, we found that both rat and human in vitro differentiation models chronically exposed to alcohol have decreased JAM-A expression at both RNA and protein levels and decreased barrier function compared to their control counterparts. Nevertheless, how chronic alcohol exposure decreases JAM-A expression and impacts airway epithelial barrier function and migration has not been fully elucidated. Chronic alcohol exposure is known to activate TGF-β1 and recent evidence has linked TGF-β1 to repression of JAM-A expression. Thus, we hypothesize that the deleterious effects of chronic alcohol exposure on conducting airway epithelial cells are due to TGF-β1 activation causing attenuation of JAM- A expression and subsequent unjamming. My research plan is designed to determine how chronic alcohol exposure (potentially through TGF-β1) impacts JAM-A expression and, therefore, barrier function in primary airway epithelial cells (Aim 1), and to define how JAM-A regulates collective cell migration/cell jamming (Aim 2). The goal of this project is to identify targetable pathways by which chronic alcohol exposure causes barrier function and cell migration defects in conducting airway epithelial cells.

项目摘要 在美国，酒精使用障碍每年影响超过 1,400 万成年人，并导致 88,000 人死亡。 长期饮酒会显着增加人们患肺部感染和急性呼吸道感染的风险 这种对损伤的敏感性增加是一种称为酒精性肺综合征的疾病， 它是由肺免疫系统和肺泡上皮屏障功能障碍引起的，但人们对此知之甚少。 关于酒精如何影响传导气道（即气管和支气管）中的上皮细胞，这是 我们使用了原代气道上皮细胞作为抵抗肺部感染性病原体的第一道防线。 从健康人和酗酒患者中分离出来，并在体外对他们进行区分，以检查酒精对他们的影响 细胞屏障功能和形态。健康分化的初级气道上皮细胞表现出鹅卵石状。 类似的模式，一旦单层成熟就变得不动——一种正常的气道细胞表型，称为 相比之下，从酒精慢性药物中分离出的气道上皮细胞仍然处于迁移状态。 “未堵塞”状态，因为单层中出现拉伸细胞和细胞漩涡的区域。 在乙醇存在下体外生长和分化的气管细胞保持畅通无阻，而对照 众所周知，跨膜蛋白连接粘附分子 A (JAM-) A) 调节上皮细胞迁移，还通过控制细胞旁细胞流动来调节屏障功能 最近，我们在大鼠和人类体外发现了小分子作为紧密连接复合物的一部分。 长期暴露于酒精的分化模型降低了 RNA 和蛋白质的 JAM-A 表达 然而，与控制措施相比，慢性酒精的水平和屏障功能下降。 暴露会降低 JAM-A 表达并影响气道上皮屏障功能和迁移尚未得到证实 已知慢性酒精暴露会激活 TGF-β1，并且最近的证据表明 TGF-β1 与此相关。 因此，我们发现了长期饮酒的有害影响。 传导气道上皮细胞的暴露是由于 TGF-β1 激活导致 JAM- 我的研究计划旨在确定酒精的慢性程度。 暴露（可能通过 TGF-β1）影响 JAM-A 表达，从而影响原发性屏障功能 气道上皮细胞（目标 1），并定义 JAM-A 如何调节集体细胞迁移/细胞干扰（目标 2）。 该项目的目标是确定长期酒精暴露导致障碍的可靶向途径 传导气道上皮细胞的功能和细胞迁移缺陷。