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 酒精使用障碍会影响超过1400万成年人，并在美国每年造成88,000人死亡。 慢性酒精使用大大增加了人们患肺部感染和急性呼吸道的风险 遇险综合征（ARDS）。对损伤的敏感性提高是一种称为酒精肺综合征的疾病， 它是由肺免疫系统和肺泡上皮屏障的功能障碍引起的。但是，鲜为人知 关于酒精如何影响导电道的上皮细胞，即气管和支气管，这是 对肺部感染性病原体的第一道防线。我们已经使用了主要气道上皮细胞 从健康和酒精患者中分离出来，并在体外区分他们，以检查酒精的影响 细胞屏障功能和形态。健康分化的原发性气道上皮细胞暴露了鹅卵石 - 像模式一样，一旦单层成熟，就变得不动 - 一种正常气道细胞表型称为 “干扰”。相比之下，从慢性酗酒者中分离出的气道上皮细胞仍在迁移 “无障碍”状态，因为单层中出现了拉伸细胞和细胞漩涡的区域。另外，大鼠 在乙醇存在下，气管细胞在体外生长和分化，而对照 单层被困住了。众所周知，跨膜蛋白连接胶分子A（jam-- a）调节上皮细胞迁移，并且还通过控制旁细胞流动来调节屏障功能 小分子作为紧密连接络合物的一部分。最近，我们发现大鼠和人类在体外都 长期暴露于酒精的分化模型在RNA和蛋白质上都改善了JAM-A的表达 与对照组相比，水平和屏障功能扩大。然而，多长期酒精 暴露会降低JAM-A表达并影响气道上皮屏障功能和迁移 已知慢性酒精暴露会激活TGF-β1，最近的证据与TGF-β1联系在一起 表达jam-a表达。那就是我们假设慢性酒精的删除效果 进行气道上皮细胞的暴露是由于TGF-β1激活引起的，导致果酱的衰减 表达和随后的无果。我的研究计划旨在确定慢性酒精 暴露（可能通过TGF-β1）会影响JAM-A表达，因此会影响原代的屏障功能 气道上皮细胞（AIM 1），并定义JAM-A如何调节集体细胞迁移/细胞干扰（AIM 2）。 该项目的目的是确定慢性酒精暴露导致障碍的目标途径 导电上皮细胞中的功能和细胞迁移缺陷。