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.

尽管在抗病毒治疗方面取得了进展，但艾滋病毒-1相关的神经疾病(HAND)已经显著 发病率增加，特别是在从事药物滥用的宿主中。可卡因是一种常见的滥用药物。 药物，并与HIV-1感染和神经发病密切相关。然而，这些的分子基础 中枢神经系统(CNS)的病理改变尚不完全清楚。最近， 细胞外小泡/外切体(EVS)与HIV-1的致病机制有关 神经炎。根据我们的初步研究，我们假设HIV-1和可卡因能增强 HIV-1感染的非法吸毒者通过改变分子载量和释放的神经发病机制 EV来自HIV-1感染的免疫细胞。这种EV有助于炎性介质和病毒的转移 通过颠覆血脑屏障(BBB)的完整性进入中枢神经系统。 本提案的总体目标是描述大型电动汽车(Levs)的分子组成。 从HIV-1感染的来源和可卡因处理的单核细胞来源的巨噬细胞，然后研究 可卡因增强EVS的细胞转运和炎症作用的机制 BBB。 具体的创新点包括：(1)利用最先进的方法来描述从 从HIV-1感染和/或可卡因处理的单核细胞来源的巨噬细胞中，(Ii)分析其生物学效应 其中对人脑微血管内皮细胞、小胶质细胞和星形胶质细胞的作用：(Iii) 探讨可卡因促进HBMECs摄取Levs的分子机制，(IV)研究 HIV-1感染巨噬细胞来源的Levs在可卡因存在或不存在时对血脑屏障的影响 (V)表征可卡因诱导的Levs中miR-34a表达增加如何增强炎症反应 HBMECs，(六)阐明所谓的“特洛伊木马病毒”，即同时具有逆转录病毒和宿主成分， 可将HIV-1感染传播给小胶质细胞和星形胶质细胞。 我们提出了三个具体目标：(1)研究可卡因对HIV-1来源的大EVS(Levs)的影响 感染的单核细胞来源的巨噬细胞，(2)探讨这些Levs对HBMEC单层的影响 有无可卡因；(3)分析可卡因对Lev诱导的炎症和HIV-1的影响 小胶质细胞和星形胶质细胞感染。 通过破译这些涉及电动汽车的分子机制，我们希望为小说提供一个框架 更有效地控制手足口病在高危人群中传播和发展的策略 可卡因吸毒者。