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 的神经病理学中的作用

• 批准号: 9788407
• 负责人: Raghava Potula
• 金额: $ 53.41万
• 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: 2018-09-30 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9788407
• 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; 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; 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

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.

抗逆转录病毒治疗（ART）的HIV感染患者的治疗有效地抑制了病毒复制。 但是，中枢神经系统（CNS）仍然是病毒的主要靶标和储层，导致该病毒 与HIV-1相关的神经认知障碍的发展（手）。重要的是，由于 艾滋病毒流行，吸毒仍然是收缩，传播和恶化的主要危险因素 艾滋病毒的结果。实际上，关于发病机理，精神刺激物可以诱导更高的病毒载荷，减少CD4 计数并提高艺术抗性的速度。此外，个体的神经心理学下降更大 是可卡因用户和艾滋病毒 - 呼吸阳性。细胞外微泡（EV），其中包括微泡 （MV）和外泌体已成为一种新型的生物学现象，几乎由每个细胞类型发布 身体。我们已经表明，脑微止血管管内皮细胞（BMVEC）衍生的电动汽车包含紧密 连接蛋白（TJP）和转运蛋白是BBB的主要成分。此外， 手的标志性特征是BBB的破坏和TJ复合物的丧失。在此提案中，我们表明 BMVEC响应HIV病毒毒素和精神刺激剂的响应。因此，我们假设艾滋病毒 滥用触发器的感染和/或药物释放EV释放，导致BBB不稳定和促进神经入侵 通过感染的免疫细胞。该提案的创新性质在三个独立目标中列出了。在 第一个目的，我们将表征EV生产程度（MV和外泌体） 精神刺激性类型，使用带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药理剂对BMVEC-EV的影响 生产。此外，我们将在体内自我管理模型以及具有或没有药物的人源化的HIV感染模型中将EC-EV释放与成瘾的阶段相关联 行政。在第二个目标中，我们介绍了BMVEC衍生的EV与激活的新颖概念 或受感染的单核细胞，这会触发单核细胞跨内皮迁移。因此，我们假设 单核细胞利用电动汽车上的TJP参与内皮紧密连接络合物并促进免疫 中枢神经系统的渗透。在第三个目标中，我们将探索一种可以最大程度地减少电动汽车生产的治疗策略 从而挽救BBB的完整性。以上将通过针对ARF6的有针对性抑制来实现 参与MV和外泌体生物发生。使用新的药理学工具抑制ARF6，我们旨在稳定 在神经炎症过程中预防BBB屏障功能障碍的BBB。本文提出的研究将提供 对脑内皮EV产生的关键见解，免疫细胞浸润到中枢神经系统的机制以及 揭示了促进HIV和药物使用中BBB保护的药理干预措施的靶标 紊乱。