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

基本信息

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
项目类别：
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

项目摘要

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.

尽管抗病毒治疗取得了进展，但 HIV-1 相关神经系统疾病 (HAND) 仍显着发病率增加，特别是在滥用可卡因的宿主中。然而，这些药物的分子基础与 HIV-1 感染和神经发病机制密切相关。近来，中枢神经系统（CNS）的病理变化尚未完全了解。细胞外囊泡/外泌体 (EV) 与 HIV-1 发病机制和引发根据我们的初步研究，我们发现 HIV-1 和可卡因会增强神经炎症。通过改变分子货物和释放的 HIV-1 感染非法药物使用个体的神经发病机制来自 HIV-1 感染的免疫细胞的 EV 促进炎症介质和病毒的转移。通过破坏血脑屏障（BBB）的完整性来影响中枢神经系统。该提案的总体目标是表征大型电动汽车（lEV）的分子成分源自 HIV-1 感染和可卡因处理的单核细胞衍生巨噬细胞，然后研究可卡因介导增强这些 EV 的转胞吞作用和炎症作用的机制 BBB。具体创新点包括：(i) 利用尖端方法来表征电动汽车的衍生特征来自 HIV-1 感染和/或可卡因处理的单核细胞衍生巨噬细胞，(ii) 分析生物效应这些 LEV 对人脑微血管内皮细胞 (HBMEC)、小胶质细胞和星形胶质细胞的影响，(iii) 探索可卡因介导的 HBMEC 增强对 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 感染的巨噬细胞的 LEV 对 BBB 的影响， (v) 描述可卡因如何诱导 lEV 中 miR-34a 的表达增加，从而增强炎症 HBMEC，(vi) 阐明所谓的“特洛伊木马 EV”是否同时拥有逆转录病毒和宿主成分，可以将 HIV-1 感染传播至小胶质细胞和星形胶质细胞。我们制定了三个具体目标：(1) 研究可卡因对 HIV-1 衍生的大型 EV（lEV）的影响感染的单核细胞衍生的巨噬细胞，(2) 探索这些 lEV 对 HBMEC 单层的影响，存在或不存在可卡因，(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，我们希望为新型电动汽车提供一个框架更有效地对抗病毒在高危人群中传播和发展的策略可卡因使用者。