Involvement of TRP Channels in Ethanol Concentration-Dependent Effects on Immune

TRP 通道参与乙醇浓度依赖性免疫效应

基本信息

批准号：
8936414
负责人：
SULIE L. CHANG
金额：
$ 15.64万
依托单位：
SETON HALL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-28 至 2018-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8936414
关键词：
Accident and Emergency department Adhesions Adolescent and Young Adult Adverse effects Affect Affinity Agonist Alcohol abuse Alcoholic Beverages Alcoholic Intoxication Alcohols Animals Beer Behavior Behavior assessment Behavioral Binding Blood Blood - brain barrier anatomy Brain C-terminal Calcium Calcium ion Cations Cell Culture Techniques Cell membrane Confocal Microscopy Consumption Data Development Endothelial Cells Ethanol Gene Expression Genes Health Hypothalamic structure Immune Immune response Immune system Immunomodulators Inflammation Inflammatory Intake Intoxication Investigation Ion Channel Ions Knock-out Knockout Mice Leukocytes Light Literature Liver Measures Mediating Metabolism Modeling Molecular Molecular Biology Techniques Molecular Conformation Molecular Profiling Molecular Structure Motor Activity Mus N-terminal NMR Spectroscopy Neurons Neurotransmitter Receptor Nuclear Magnetic Resonance Patients Pattern Permeability Phorbol Phorbols Physiological Play Population Process Production Protein Conformation Proteins Rattus Reporting Research Resolution Role Social Problems Solutions Spleen Stimulus Structure TRPA1 Channel TRPV1 gene Taste Perception Techniques Temperature Sense Testing Therapeutic Time Transmembrane Domain Vascular Endothelium Vasopressins Wild Type Mouse Wine alcohol content base binge drinking cell type cellular targeting chemokine clinically relevant cytokine distilled alcoholic beverage drinking extracellular innovation intravital microscopy prevent psychologic receptor receptor expression response structural biology supraoptic nucleus young adult

项目摘要

DESCRIPTION (provided by applicant): Binge drinking, or the consumption of a large volume of alcoholic beverages in a very short time, is a serious social problem, particularly among adolescents and young adults. Ethanol (EtOH), the main ingredient in alcoholic beverages, is well known for its behavioral and psychological effects and also as an immune system modulator. However, the underlying mechanisms by which EtOH exerts its various effects are still not defined. Although recent evidence indicates that EtOH acts at the cellular level, there i still a fundamental gap between EtOH's potential cellular targets and its subsequent physiological effects. One such target is a group of cell membrane ion channels called transient receptor potential (TRP) channels. TRP channels are found in various cell types, including brain microvascular endothelial cells (BMVEC) of the blood-brain barrier (BBB). TRP channels mediate certain immune responses, such as cytokine production and leukocyte-endothelial adhesion (LEA), the initial step in the inflammatory process. Activation of TRP channels by various extracellular stimuli, including EtOH, induces an influx of calcium ions, which can subsequently increase LEA. Alcoholic beverages differ in their EtOH content or alcohol-by-volume (ABV) concentration, and EtOH's effects appear to be concentration dependent. We recently reported that binge consumption of solutions with high EtOH concentrations causes more pronounced immune responses than those with low EtOH concentrations, even when the amount of EtOH intake is the same. Based on the recent literature and our preliminary studies, we hypothesize that TRP channels mediate alcohol- induced immune responses at the blood-brain barrier (BBB) in an EtOH concentration- dependent manner. To test our hypothesis, we propose the following two specific aims: (1) To determine the effects of EtOH concentration on the structural binding of EtOH to TRPV4 channels in BMVEC using NMR spectroscopic techniques; and (2) To delineate the involvement of TRPV4 channels in the EtOH concentration-dependent effects on immune responses in the BMVEC at the BBB. In this application, we will combine investigation of the structural biology of EtOH-protein interactions with examination of EtOH's effects in animal and cell culture models to determine the mechanisms by which EtOH affects immune responses. This study is innovative because, to our knowledge, few studies have examined the relationship between EtOH-protein interaction and EtOH-induced immune effects at the level of the vascular endothelium. The NMR structural studies will provide essential details concerning EtOH-TRP binding and the EtOH concentration-dependent changes in target protein conformation. The molecular studies will then determine the correlation between those changes and the immunomodulatory effects of EtOH. Our study is highly significant and clinically relevant because it will provide valuable information on the mechanisms underlying the physiological effects of binge drinking with high ABV alcoholic beverages, which can help to prevent EtOH-induced dysregulation of immune responses and be used to develop therapeutic strategies to treat patients with alcoholic intoxication seen in the emergency room.

描述（由申请人提供）：酗酒，即在很短的时间内饮用大量酒精饮料，是一个严重的社会问题，特别是在青少年和年轻人中。乙醇 (EtOH) 是酒精饮料的主要成分，因其行为和心理影响以及免疫系统调节剂而闻名。然而，乙醇发挥其各种作用的潜在机制仍不清楚。尽管最近的证据表明乙醇在细胞水平上起作用，但乙醇的潜在细胞靶标与其随后的生理效应之间仍然存在根本差距。其中一个目标是一组细胞膜离子通道，称为瞬时受体电位 (TRP) 通道。 TRP 通道存在于多种细胞类型中，包括血脑屏障 (BBB) 的脑微血管内皮细胞 (BMVEC)。 TRP 通道介导某些免疫反应，例如细胞因子的产生和白细胞内皮粘附 (LEA)，这是炎症过程的第一步。各种细胞外刺激（包括 EtOH）激活 TRP 通道会诱导钙离子流入，从而导致 LEA 增加。酒精饮料的乙醇含量或酒精体积 (ABV) 浓度各不相同，并且乙醇的效果似乎与浓度相关。我们最近报道，即使乙醇摄入量相同，暴饮暴食高浓度乙醇溶液比低浓度乙醇溶液引起更明显的免疫反应。根据最近的文献和我们的初步研究，我们假设 TRP 通道以 EtOH 浓度依赖性方式介导血脑屏障 (BBB) 上酒精诱导的免疫反应。为了检验我们的假设，我们提出以下两个具体目标：（1）使用核磁共振波谱技术确定EtOH浓度对BMVEC中EtOH与TRPV4通道结构结合的影响； (2) 描述 TRPV4 通道对 BBB 处 BMVEC 免疫反应的 EtOH 浓度依赖性影响的参与。在此应用中，我们将把乙醇-蛋白质相互作用的结构生物学研究与乙醇在动物和细胞培养模型中的影响相结合，以确定乙醇影响免疫反应的机制。这项研究具有创新性，因为据我们所知，很少有研究探讨乙醇-蛋白质相互作用与乙醇诱导的血管内皮水平免疫效应之间的关系。 NMR 结构研究将提供有关 EtOH-TRP 结合和靶蛋白构象中 EtOH 浓度依赖性变化的重要细节。然后分子研究将确定这些变化与乙醇的免疫调节作用之间的相关性。我们的研究具有非常重要的意义和临床相关性，因为它将提供关于酗酒高 ABV 酒精饮料的生理影响机制的有价值的信息，这有助于预防乙醇引起的免疫反应失调，并可用于制定治疗策略治疗急诊室出现的酒精中毒患者。