Injury of blood brain and alveolar-endothelial barriers caused by alcohol and electronic cigarettes via purinergic receptor signaling

酒精和电子烟通过嘌呤受体信号传导引起血脑和肺泡内皮屏障损伤

基本信息

批准号：
10638221
负责人：
Yuri Persidsky
金额：
$ 59.49万
依托单位：
TEMPLE UNIV OF THE COMMONWEALTH
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2028-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10638221
关键词：
3-Dimensional Adhesions Adolescent Adverse effects Affect Alcohols Alveolar Animal Model Animals Bacterial Infections Biological Assay Blood Blood - brain barrier anatomy Blood Vessels Brain Brain Injuries Bronchoalveolar Lavage C-reactive protein Cells Cerebrovascular Disorders Chronic Clinical Research Cognition Cognitive Data Devices Dissection Electronic cigarette Endothelial Cells Endothelium Epithelial Cells Evaluation Exposure to Functional disorder Generations Glucose Transporter Health Hippocampus Human Immune response Immune system Immunohistochemistry Impaired cognition Impairment In Vitro Individual Infiltration Inflammation Inflammatory Inflammatory Response Inhalation Injury Knockout Mice Knowledge Leukocytes Link Lipids Long-Term Effects Lung Macrophage Magnetic Resonance Imaging Marketing Measures Mediating Messenger RNA Metals Microglia Mitochondria Mus Myosin Light Chain Kinase Nicotine Organ Oxidative Stress P2X-receptor Pathology Pathway interactions Permeability Phenotype Phosphotransferases Play Production Proteins Publications Purinoceptor Reaction Reactive Oxygen Species Receptor Signaling Rho-associated kinase Role Safety Secondary to Smoking Structure of parenchyma of lung System Testing Therapeutic Intervention Tight Junctions Virus Diseases Work alcohol exposure alcohol use disorder antagonist blood-brain barrier disruption blood-brain barrier function blood-brain barrier permeabilization brain endothelial cell cellular targeting central nervous system injury cerebral microvasculature cigarette smoke cognitive function combustible tobacco cytokine e-cigarette aerosols electronic cigarette use electronic cigarette user endothelial dysfunction experimental study extracellular feeding immune activation improved in vivo inflammatory marker innovation intercellular cell adhesion molecule knockout animal lung injury mitochondrial dysfunction multidrug abuse neuroinflammation neurotoxic neurotrophic factor neutrophil prevent receptor response systemic inflammatory response tissue injury vaping volunteer

项目摘要

Polydrug abuse (especially alcohol use disorder, AUD, and smoking) are known individually to compromise the lung alveolar-endothelial barrier (AEB) and the blood brain barrier (BBB). Very limited knowledge exists regarding damage in lung and brain due to electronic cigarettes (e-Cig) in combination with AUD. E-Cig have become popular, yet very limited data indicate that they cause endothelial dysfunction and result in a pro-inflammatory phenotype in macrophages and endothelium in lungs. While e-Cig are known to be addictive, their effects on the brain and cognition are essentially unknown. Our data show that chronic e-Cig exposure in mice enhanced permeability of the BBB and neuroinflammation, diminished expression of a key glucose transporter and tight junction protein on brain endothelium, and impaired cognition. Preliminary data indicate that the combination of alcohol/e-Cig exposure in vivo caused enhanced AEB permeability and amplified neuroinflammation/BBB compromise. We found that e-Cig and alcohol impair AEB and BBB via the same mechanism including mitochondrial dysfunction, Ca2+ accumulation, and ATP extracellular release, potentially mediated by purinergic receptor, P2X7, in cellular components of AEB/BBB. Using innovative in vitro 3D systems of AEB and BBB and relevant animal models, we test the hypothesis that BBB and AEB injury in e-Cig/alcohol exposure are mediated through the P2X7 receptor. In aim 1, we will screen the magnitude of injury (mitochondrial dysfunction, Ca2+ increase and ATP release) by various types of e-Cig in combination with alcohol on human brain and lung endothelial and lung epithelial cells. Then, we will define mechanisms of demise using innovative 3D in vitro constructs of lung and brain microvasculature, functional assays, assessment of mitochondrial functions and expression of key molecules supporting BBB and AEB. We will investigate the contribution of activation of the purinergic P2X7 receptor in e-Cig/alcohol induced BBB/AEB dysfunction. The 2nd aim will study in vivo lung injury after chronic alcohol feeding and e-Cig vaping evaluating AEB permeability, expression of barrier supporting molecules, inflammatory responses (immunohistochemistry, protein/mRNA, bronchoalveolar lavage). P2X7 knockout (KO) animals will allow dissection of the role of this receptor in pulmonary dysfunction. The 3rd aim will decipher combined in vivo effects of BBB function, expression of barrier-mediating molecules, and neuroinflammation. The same experiments performed in P2X7 KO mice will determine the importance of this pathway in CNS injury. Markers of lung injury and BBB damage will be measured in blood and correlated with signs of end-organ pathology.

多药滥用（尤其是酒精使用障碍，AUD和吸烟）是单独知道的妥协肺肺泡 - 内皮屏障（AEB）和血脑屏障（BBB）。非常关于电子卷烟引起的肺和大脑损害的知识有限（E-cig）结合AUD。电子烟已经流行，但数据非常有限，表明他们引起内皮功能障碍，并导致巨噬细胞中的促炎表型肺内皮。虽然众所周知电子烟是令人上瘾的，但它们对大脑的影响认知本质上是未知的。我们的数据表明，小鼠的慢性电子烟暴露 BBB和神经炎症的渗透性增强，钥匙的表达降低葡萄糖转运蛋白和紧密连接蛋白在脑内皮上，认知受损。初步数据表明，酒精/电子烟暴露在体内引起的结合增强了AEB渗透性和扩增的神经炎症/BBB折衷。我们发现电子烟和酒精通过包括线粒体在内的相同机制损害了AEB和BBB 功能障碍，Ca2+积累和ATP细胞外释放，可能由嘌呤能受体P2X7，在AEB/BBB的细胞成分中。使用创新的体外3D AEB和BBB的系统以及相关的动物模型，我们检验了BBB和通过P2X7受体介导E-CIG/酒精暴露中的AEB损伤。在AIM 1中，我们将通过筛选损伤的幅度（线粒体功能障碍，Ca2+增加和ATP释放）各种类型的电子烟与人脑上的酒精结合使用，肺内皮和肺上皮细胞。然后，我们将使用创新的3D体外定义灭亡机制肺和脑微举行的构建体，功能测定，线粒体评估支持BBB和AEB的关键分子的功能和表达。我们将调查 E-CIG/酒精诱导的BBB/AEB激活嘌呤能P2X7受体的激活的贡献功能障碍。第二个目标将在慢性酒精喂养和电子烟后研究体内肺损伤评估AEB渗透性的烟雾，屏障支撑分子的表达，炎症反应（免疫组织化学，蛋白质/mRNA，支气管肺泡灌洗）。 P2X7淘汰赛（KO）动物将允许解剖该受体在肺功能障碍中的作用。第三 AIM将破译BBB功能的体内效应，表达屏障介绍分子和神经炎症。在P2X7 KO小鼠中进行的相同实验将确定该途径在中枢神经系统损伤中的重要性。肺损伤和BBB的标记损害将在血液中测量，并与最终器官病理学的迹象相关。