Opioids, Extracellular Vesicles and BBB Innate Immunity

摘要 细胞外小泡(EVS)在神经元存活和中枢免疫监视中发挥着重要作用。 神经系统(CNS)通过调节神经元和其他脑细胞之间的细胞间通讯而发挥作用。我们的早期 体外和体内研究表明，阿片类药物(吗啡和海洛因)和/或HIV可以抑制a基因的表达。 细胞内和循环中的HIV限制性微RNA(MiRNAs)的数量。最近，我们展示了 EVS可以将免疫激活的脑微血管内皮细胞(BMVEC)的抗病毒信号运送到 巨噬细胞，导致艾滋病毒抑制。此外，我们还发现，来自HIV感染者的血浆EV 富含几种具有神经毒性作用的miRNA(miR-21和let-7)或靶向紧密连接蛋白(miR-17和 MIR-20a)。因此，我们假设阿片类药物和/或艾滋病毒损害了血脑屏障(BBB)的完整性，并使 EVS介导的miRNAs细胞间传递的神经毒性。我们提出了两个具体目标来解决 这一假设：具体目的：1.确定EVS介导的miRNAs对BBB先天免疫的影响 HIV与阿片类药物使用背景下的神经元损伤；具体目标2.从机制上研究BMVEC- 而血浆衍生的EV在血脑屏障的先天抗病毒免疫和神经元损伤方面，我们将产生生物工程 包含特定miRNAs的外体。然后我们将确定这些miRNAs的3‘UTR靶标，以及 这些miRNAs参与了BMVEC和神经细胞的免疫激活和炎症的调节。 该项目的发现将为EVS介导的血脑屏障对艾滋病毒和艾滋病的天然免疫提供科学证据 阿片类药物和/或HIV介导的神经元损伤的新机制。

Abstract Extracellular vesicles (EVs) play an important role in neuronal survival and immune surveillance of the central nervous system (CNS) by mediating the intercellular communication between neurons and other brain cells. Our early in vitro and in vivo studies showed that opioids (morphine and heroin) and/or HIV could inhibit the expression of a number of the intracellular and circulating HIV restriction microRNAs (miRNAs). Recently, we demonstrated that EVs could shuttle antiviral signals from immune-activated brain microvascular endothelial cells (BMVEC) to macrophages, resulting in HIV inhibition. In addition, we showed that plasma EVs from HIV-infected subjects enriched several miRNAs that have neurotoxic effect (miR-21 and let-7) or target tight junction proteins (miR-17 and miR-20a). We thus hypothesize that opioids and/or HIV impair the blood-brain barrier (BBB) integrity and exert neurotoxicity via EVs-mediated intercellular transmission of miRNAs. We propose two specific aims to address this hypothesis: Specific Aim 1. To determine the impact of EVs-mediated miRNAs on BBB innate immunity against HIV and neuronal injury in the context of opioid use; Specific Aim 2. To mechanistically study the role of BMVEC- and plasma-derived EVs in the BBB innate antiviral immunity and neuronal injury, we will generate bioengineered exosomes that contain the specific miRNAs. We will then determine the 3’ UTR targets of these miRNAs and whether these miRNAs are involved in the modulation of immune activation and inflammation in BMVEC and neuronal cells. The findings from this project will provide scientific evidence of EVs-mediated BBB innate immunity against HIV and a novel mechanism for opioid and/or HIV-mediated neuronal injury.