Cocaine induces production of infectious large extracellular vesicles (lEV) and regulates neuro-inflammation

可卡因诱导产生传染性大细胞外囊泡 (lEV) 并调节神经炎症

• 批准号: 10208847
• 负责人: JEROME E GROOPMAN
• 金额: $ 38.37万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-15 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10208847
• 关键词: Acquired Immunodeficiency Syndrome; Address; Anti-Inflammatory Agents; Antiviral Therapy; Astrocytes; Binding; Biological; Biological Process; Blood - brain barrier anatomy; Brain; Cell Communication; Cells; Cocaine; Cocaine Users; Communication; Development; Fostering; HIV; HIV-1; Heterogeneity; Human; Immune; Incidence; Individual; Infection; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; Mediating; Membrane; Methodology; MicroRNAs; Microglia; Molecular; Neuraxis; Neuropathogenesis; Pathogenesis; Pathologic; Pathway interactions; Play; Production; Reaction; Role; Signal Pathway; Substance abuse problem; Techniques; Viral; Virus; Virus Replication; axl receptor tyrosine kinase; base; blood-brain barrier permeabilization; brain endothelial cell; cofactor; combat; cytokine; design; drug of abuse; exosome; extracellular; extracellular vesicles; high risk behavior; high risk population; illicit drug use; innovation; insight; macrophage; monocyte; monolayer; nanoparticle; nanosized; nervous system disorder; neuroinflammation; novel; novel strategies; particle; prevent; response; transcytosis; transmission process; uptake; vesicular release

Despite advances in antiviral therapy, HIV-1 associated neurological disorders (HAND) have significantly increased in incidence, particularly in hosts who engage in substance abuse. Cocaine is a commonly abused drug and strongly implicated in HIV-1 infection and neuropathogenesis. However, the molecular basis of these pathological changes in the central nervous system (CNS) is still not yet fully understood. Recently, extracellular vesicles/exosomes (EVs) have been implicated in HIV-1 pathogenesis and elicitation of neuroinflammation. Based on our preliminary studies, we hypothesize that HIV-1 and cocaine enhance neuropathogenesis in HIV-1 infected illicit drug using individuals by altering the molecular cargo and release of EVs from HIV-1 infected immune cells. Such EVs facilitate the transfer of inflammatory mediators and viral components to the CNS by subverting the integrity of the blood brain barrier (BBB). The overall objective of this proposal is to characterize the molecular components of large EVs (lEVs) derived from HIV-1 infected and cocaine treated monocyte-derived macrophages, and then study the mechanisms involved in cocaine mediated enhanced transcytosis and inflammatory effects of these EVs on the BBB. Specific points of innovation include: (i) utilize cutting edge methodologies to characterize the lEVs derived from HIV-1 infected and/or cocaine treated monocyte-derived macrophages, (ii) analyze the biological effects of these lEVs on human brain microvascular endothelial cells (HBMECs), microglial cells and astrocytes, (iii) explore molecular mechanisms involved in cocaine mediated enhanced uptake of lEVs by HBMECs , (iv) study the effects of lEVs derived from HIV-1 infected macrophages on BBB, in the presence or absence of cocaine, (v) characterize how cocaine induced increased expression of miR-34a in lEVs enhances inflammation in HBMECs, (vi) elucidate whether so-called “ Trojan EVs”, that possess both retroviral and host components, can transmit HIV-1 infection to microglia and astrocytes. We set out three specific aims to: (1) Study the effects of cocaine on large EVs (lEVs) derived from HIV-1 infected monocyte-derived macrophages, (2) Explore the effects of these lEVs on a HBMEC monolayer, in the presence or absence of cocaine, (3) Analyze the effects of cocaine on lEV induced inflammation and HIV-1 infection in microglial cells and astrocytes. By deciphering these molecular mechanisms involving EVs, we hope to provide a framework for novel strategies to more effectively combat virus spread and development of HAND in the high risk population of cocaine users.

尽管在抗病毒疗法方面取得了进步，但HIV-1相关的神经系统疾病（手）显着 发病率有所增加，特别是在从事滥用药物的宿主中。可卡因通常是滥用的 药物，与HIV-1感染和神经病发生密切相关。但是，这些分子基础 中枢神经系统（CNS）的病理变化尚未完全了解。最近， 在HIV-1发病机理中隐含了细胞外蔬菜/外泌体（EV） 神经炎症。根据我们的初步研究，我们假设HIV-1和可卡因增强了 通过改变分子货物和释放，使用个体的HIV-1感染非法药物的神经病发生 来自HIV-1感染的免疫细胞的电动汽车。这样的电动汽车促进了炎症介质和病毒的转移 通过颠覆血脑屏障（BBB）的完整性，将CNS的组成部分。 该提案的总体目的是表征大电动汽车（LEV）的分子成分 源自受HIV-1感染和可卡因处理的单核细胞衍生的巨噬细胞，然后研究 可卡因介导的增强的转胞病和这些电动汽车的炎症作用涉及的机制 BBB。 创新的特定点包括：（i）利用尖端方法来表征衍生的LEV 从感染HIV-1和/或可卡因处理的单核细胞衍生的巨噬细胞中，（ii）分析生物学作用 这些在人脑微血管内皮细胞（HBMEC），小胶质细胞和星形胶质细胞的LEV中，（iii） 探索与可卡因介导的HBMEC涉及的LEV摄取的分子机制，（iv）研究 在存在或不存在可卡因的情况下，来自HIV-1感染巨噬细胞的LEV对BBB的影响， （v）表征可卡因如何诱导LEV中miR-34a的表达增加会增强炎症 HBMecs（vi）阐明是否具有逆转录病毒和宿主组件的所谓“特洛伊电动汽车”， 可以将HIV-1感染传输到小胶质细胞和星形胶质细胞。 我们设定了三个特定的目的：（1）研究可卡因对源自HIV-1的大型电动汽车（LEV）的影响 感染单核细胞衍生的巨噬细胞，（2）探索这些LEV对HBMEC单层的影响， 可卡因的存在或不存在，（3）分析可卡因对LEV诱导注射和HIV-1的影响 小胶质细胞和星形胶质细胞的感染。 通过破译这些涉及电动汽车的分子机制，我们希望为新颖的框架提供一个框架 在高风险人群中更有效地战斗病毒传播和开发手的策略 可卡因用户。