Brain endothelial EVs role in the neuropathology of drugs of abuse and HIV

脑内皮细胞 EV 在滥用药物和 HIV 神经病理学中的作用

基本信息

批准号：
9930925
负责人：
Raghava Potula
金额：
$ 0.85万
依托单位：
TEMPLE UNIV OF THE COMMONWEALTH
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-30 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9930925
关键词：
Affect Animal Model Attenuated Autopsy Bathing Binding Biogenesis Biological Phenomena Blood - brain barrier anatomy Brain CD4 Lymphocyte Count Carrier Proteins Cell model Cells Cerebrovascular system Chronic Cocaine Users Complex Contracts Data Development Dose Drug Exposure Drug Modelings Drug usage Endothelial Cells Endothelium Epidemic Equus caballus Event Exhibits Exposure to Extracellular Protein Extravasation FCGR3B gene Functional disorder HIV HIV Infections HIV Seropositivity HIV-associated neurocognitive disorder Homeostasis Human Immune In Vitro Individual Infiltration Influentials Intervention Kinetics Laboratories Methamphetamine Microfluidics Modeling Monomeric GTP-Binding Proteins Mus Nature Neuraxis Neuropathogenesis Neuropsychology Outcome Pathogenesis Patients Pharmaceutical Preparations Pharmacology Phase Production Proteins Reporting Resistance Risk Factors Role Salts Self Administration Signaling Molecule Substance Use Disorder System Testing Therapeutic Tight Junctions Tissues United States National Institutes of Health Viral Load result Viral reservoir Virus Replication addiction antiretroviral therapy brain endothelial cell cell motility cell type cerebral microvasculature cocaine exposure drug of abuse exosome extracellular extracellular vesicles humanized mouse in vivo inhibitor/antagonist innovation insight microvesicles migration monocyte mouse model negative affect neuroinflammation neuropathology neurovascular unit novel novel therapeutics prevent psychostimulant response tool virotoxins virtual

项目摘要

Treatment of HIV-infected patients with antiretroviral therapy (ART) has effectively suppressed viral replication; however, the central nervous system (CNS) is still a major target and reservoir of the virus leading to the development of HIV-1-associated neurocognitive disorders (HAND). Importantly, since the beginning of the HIV epidemic, drug use has remained a primary risk factor for contracting, transmitting, and worsening the outcomes of HIV. In fact, regarding pathogenesis, psychostimulants can induce higher viral loads, reduce CD4 counts, and increase rates of ART resistance. Furthermore, neuropsychological decline is greater in individuals that are cocaine users and HIV-seropositive. Extracellular microvesicles (EVs), which includes microvesicles (MVs) and exosomes, have emerged as a novel biological phenomenon, released by virtually every cell type in the body. We have shown that brain microvasculature endothelial cell (BMVEC)-derived EVs contain tight junction proteins (TJPs) and transporter proteins, which are main constituents of the BBB. Furthermore, a hallmark feature of HAND is the disruption of the BBB and loss of TJ complexes. In this proposal, we show that BMVECs shed EVs in response to HIV virotoxins and psychostimulants. Thus, we hypothesize that HIV infection and/or drugs of abuse triggers EV release leading to BBB instability and facilitation of neuroinvasion by infected immune cells. The innovative nature of this proposal is featured in three independent aims. In the first aim, we will characterize the degree of EV production (MVs and exosomes) as a function of psychostimulant type using a novel microfluidic model of the neurovascular unit with primary human cells. We will also investigate the effects of exposure to HIV virotoxins and ART pharmacologic agents on BMVEC-EV production. Furthermore, we will correlate EC-EV release to the phases of addiction in an in-vivo self-administration model as well as in a humanized mouse model of HIV infection with or without drug administration. In the second aim, we introduce the novel concept that BMVEC-derived EVs bind to activated or infected monocytes, which triggers increased monocytic transendothelial migration. Thus, we hypothesize that monocytes utilize TJPs on EVs to engage endothelial tight junction complexes and facilitate immune infiltration of the CNS. In the third aim, we will explore a therapeutic strategy that could minimize EV production and thus rescue BBB integrity. The above will be accomplished by targeted inhibition of ARF6, which is involved in MV and exosome biogenesis. Using new pharmacological tools to inhibit ARF6, we aim to stabilize the BBB to prevent BBB barrier dysfunction during neuroinflammation. The studies proposed herein will offer crucial insight to brain endothelial EV production, mechanisms of immune cell infiltration into the CNS, and also reveal targets for pharmacological interventions that promote BBB protection in HIV and substance use disorder.

对HIV感染者进行抗逆转录病毒疗法（ART）治疗，可有效抑制病毒复制；然而，中枢神经系统（CNS）仍然是病毒的主要目标和储存库，导致 HIV-1 相关神经认知障碍 (HAND) 的发展。重要的是，自年初以来艾滋病毒流行、吸毒仍然是感染、传播和恶化艾滋病毒的主要危险因素艾滋病毒的结果。事实上，就发病机制而言，精神兴奋剂可以诱导更高的病毒载量，减少CD4 计数，并增加 ART 耐药率。此外，个体的神经心理衰退更为严重他们是可卡因使用者且艾滋病毒血清呈阳性。细胞外微泡 (EV)，包括微泡（MV）和外泌体，已成为一种新的生物现象，几乎由所有类型的细胞释放身体。我们已经证明，脑微血管内皮细胞 (BMVEC) 衍生的 EV 含有紧密的连接蛋白（TJP）和转运蛋白，它们是血脑屏障的主要成分。此外，一个 HAND 的标志性特征是 BBB 的破坏和 TJ 复合体的丢失。在这个提案中，我们表明 BMVEC 响应 HIV 病毒毒素和精神兴奋剂而释放 EV。因此，我们假设艾滋病毒感染和/或滥用药物触发 EV 释放，导致 BBB 不稳定并促进神经侵袭被感染的免疫细胞。该提案的创新性体现在三个独立的目标上。在第一个目标，我们将描述 EV 产生的程度（MV 和外泌体）作为精神兴奋剂类型使用具有原代人类细胞的神经血管单元的新型微流体模型。我们还将研究暴露于 HIV 病毒毒素和 ART 药物对 BMVEC-EV 的影响生产。此外，我们将在体内自我给药模型以及使用或不使用药物的 HIV 感染人源化小鼠模型中将 EC-EV 释放与成瘾阶段相关联行政。在第二个目标中，我们引入了 BMVEC 衍生的 EV 与激活的或感染的单核细胞，这会引发单核细胞跨内皮迁移增加。因此，我们假设单核细胞利用 EV 上的 TJP 来接合内皮紧密连接复合物并促进免疫中枢神经系统的浸润。在第三个目标中，我们将探索一种可以最大限度地减少电动汽车产量的治疗策略从而挽救 BBB 完整性。上述工作将通过靶向抑制 ARF6 来实现，即参与 MV 和外泌体生物合成。使用新的药理学工具抑制 ARF6，我们的目标是稳定 BBB 可以防止神经炎症期间 BBB 屏障功能障碍。本文提出的研究将提供对脑内皮细胞 EV 产生、免疫细胞浸润到中枢神经系统的机制的重要见解，以及揭示促进 HIV 和物质使用中 BBB 保护的药物干预目标紊乱。