Chemokine CXCL12/CXCR4 system and synthetic cathinones

趋化因子CXCL12/CXCR4系统和合成卡西酮

• 批准号: 10187189
• 负责人: SCOTT M. RAWLS
• 金额: $ 15.85万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10187189
• 关键词: 2019-nCoV; Acute; Adult; Area; Award; Behavior; Biological Response Modifiers; Blood - brain barrier anatomy; Brain; Brain region; COVID-19; COVID-19 pandemic; CXCL12 gene; CXCR4 gene; Cessation of life; Chemosensitization; Chronic; Cocaine; Complement; Contracts; Coronavirus; Cytoskeletal Proteins; Cytoskeleton; Electrical Resistance; Endothelial Cells; Endothelium; Enterocytes; Epithelial Cells; Exposure to; Female; Fluorescence Microscopy; Functional disorder; Funding; Human; In Vitro; Inflammation Mediators; Inflammatory; Inflammatory Response; Investigation; Laboratories; Lung; Measurement; Measures; Microbe; Microscopy; Mitochondria; Molecular; Neuraxis; Nucleus Accumbens; Parents; Peptidyl-Dipeptidase A; Permeability; Persons; Prefrontal Cortex; Proteins; Rattus; Reactive Oxygen Species; Research; Resistance; Rewards; Role; Series; Small Intestines; Surface; System; Testing; Tight Junctions; Time; Toxic effect; Toxin; Ventral Tegmental Area; Virus; alveolar epithelium; awake; bath salts; blood damage; blood-brain barrier function; blood-brain barrier permeabilization; brain endothelial cell; brain tissue; chemokine; cocaine exposure; cocaine use; cytokine; drug seeking behavior; experimental study; fluorescence microscope; frontal lobe; in vivo; insight; lung injury; male; monolayer; neuroinflammation; novel; psychostimulant; receptor; response; time use

Human death in persons infected with SARS-CoV-2 (the virus responsible for the covid-19 pandemic) is caused by barriers dysfunction (i.e. in the pulmonary system). Angiotensin- converting enzyme 2 (ACE2) has been identified as a functional receptor of SARS-CoV-2, similarly to other coronaviruses. Surface expression of ACE2 protein was found on lung alveolar epithelial cells and enterocytes of the small intestine. In brain, ACE2 is expressed on endothelial cells, a major component of blood-brain barrier (BBB). We propose to perform a series of studies to assess the impact of SARS-CoV-2 spike protein on BBB function in the presence of cocaine as an extension of our funded research on psychostimulants and the CXCL12/CXCR4 chemokine system. First, a comprehensive series of experiments on the impact of SARS-CoV-2 spike protein in the presence and absence of cocaine on brain microvascular endothelial cells will be performed in vitro to inform on the cellular and molecular mechanism involved in altered BBB function. Included in the analysis will be measurements of cytosolic Ca2+ levels, mitochondrial reactive oxygen species, tight junctions and cytoskeletal proteins. Second, integrated fluorescence microscopy will be used to visualize and quantify changes in BBB permeability in real time in awake rats after exposure to SARS-CoV-2 spike protein. The impact of acute and chronic administration of cocaine on BBB function in the setting of SARS-CoV-2 spike protein will be determined. Finally, brain regions from rats exposed to cocaine and the spike protein will be examined for levels of pro-inflammatory cytokines and chemokines. Taken together, this series of experiments will provide novel information about the effect of the spike protein on BBB function and neuroinflammation, and its potential interactions with cocaine. We hypothesize that chronic cocaine exposure will exacerbate the negative impact of SARS-CoV-2 spike protein on BBB integrity and increase pro-inflammatory mediators in the CNS. These studies will provide the necessary groundwork to embark on a larger study of the impact of SARS-CoV-2 on cocaine behaviors and toxicities.

感染SARS-CoV-2(导致新冠肺炎的病毒)的人死亡 大流行)是由屏障功能障碍(即在肺部系统)引起的。血管紧张素- 转换酶2(ACE2)是SARS-CoV-2的功能性受体， 与其他冠状病毒相似。在肺组织中发现ACE2蛋白的表面表达 小肠的肺泡上皮细胞和肠细胞。在大脑中，ACE2表达于 内皮细胞是血脑屏障(BBB)的主要组成部分。我们建议执行一项 评估SARS-CoV-2刺突蛋白对儿童血脑屏障功能影响的系列研究 可卡因的存在是我们资助的精神刺激剂和 CXCL12/CXCR4趋化因子系统。首先，进行了一系列关于 SARS-CoV-2刺突蛋白在可卡因存在和不存在时对脑的影响 微血管内皮细胞将在体外进行，以告知细胞和分子 血脑屏障功能改变的机制。分析中将包括以下测量 细胞内钙离子水平、线粒体活性氧、紧密连接和细胞骨架 蛋白质。第二，将使用集成荧光显微镜进行可视化和量化 SARS-CoV-2尖峰暴露后清醒大鼠血脑屏障通透性的实时变化 蛋白。急、慢性使用可卡因对老年人血脑屏障功能的影响 SARS-CoV-2刺突蛋白的设定将被确定。最后，老鼠的大脑区域 暴露在可卡因中，刺激性蛋白将被检查促炎症水平 细胞因子和趋化因子。综上所述，这一系列实验将提供新颖的 有关Spike蛋白对血脑屏障功能和神经炎症的影响的信息，以及其 与可卡因的潜在相互作用。我们假设长期接触可卡因会 加剧SARS-CoV-2刺突蛋白对血脑屏障完整性和增加的负面影响 中枢神经系统中的促炎介质。这些研究将为以下工作提供必要的基础 就SARS-CoV-2对可卡因行为和毒性的影响展开更大规模的研究。