Opioids, Extracellular Vesicles and BBB Innate Immunity

摘要 细胞外囊泡（EV）在神经元存活和中枢神经系统的免疫监视中起重要作用。 通过介导神经元和其他脑细胞之间的细胞间通讯，在神经系统（CNS）中发挥作用。我们早期 体外和体内研究表明，阿片类药物（吗啡和海洛因）和/或艾滋病毒可以抑制一种 细胞内和循环中HIV限制性microRNA（miRNAs）的数量。最近，我们证明， EV可以将免疫激活的脑微血管内皮细胞（BMVEC）的抗病毒信号穿梭于 巨噬细胞，导致HIV抑制。此外，我们发现，来自HIV感染者的血浆EV 富集了具有神经毒性作用（miR-21和let-7）或靶向紧密连接蛋白（miR-17和let-7）的几种miRNA， miR-20a）。因此，我们假设阿片类药物和/或艾滋病毒损害血脑屏障（BBB）的完整性，并发挥 通过EV介导的miRNA细胞间传递的神经毒性。我们提出两个具体目标， 具体目标1。确定EV介导的miRNA对BBB先天免疫的影响 阿片类药物使用背景下的艾滋病毒和神经元损伤;具体目标2。从机制上研究BMVEC的作用- 以及血浆来源的EV在BBB先天性抗病毒免疫和神经元损伤中的作用，我们将产生生物工程 含有特定miRNA的外泌体。然后我们将确定这些miRNAs的3' UTR靶点， 这些miRNA参与BMVEC和神经元细胞中免疫活化和炎症的调节。 该项目的发现将为EV介导的BBB先天免疫对抗HIV提供科学证据， 阿片和/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.