Role of the NLRP3 Inflammasome in Mediating Microglial Activation and Development of Neuropsychiatric Symptoms Induced by HIV, Antiretrovirals and Cocaine

摘要 尽管艾滋病毒感染者的预期寿命与艾滋病毒阴性人群相当， 尽管如此，由于神经精神疾病的流行，他们的生活质量仍然受到严重影响，其中包括 抑郁和焦虑这些人大脑中的主要病理变化包括异常的小胶质细胞 (Mg)激活和神经元损伤。药物滥用是艾滋病毒感染的一种高并发症，滥用药物可能 夸大了PLWHIV中现有的神经系统并发症。这种现象背后的详细机制 现象仍然非常难以捉摸。已经提出了几个促成因素，这种神经 包括持续表达HIV蛋白如转录反式激活因子（达特）、 长期使用抗逆转录病毒药物和滥用药物。我们的初步数据表明：1）NLRP 3 炎症体信号参与HIV-TAT介导的Mg激活; 2）可卡因通过 miR-124/TLR 4轴失调，这可以通过抑制NLRP 3炎性体来逆转; 3） 临床实践中使用的联合抗逆转录病毒药物（替诺福韦：TFV，恩曲他滨：FTC和度鲁特韦：DTG）可以 通过溶酶体功能障碍激活Mg; 4）Mg共同暴露于三种药物（达特、可卡因和ARV） 有趣的是，导致体外NLRP 3炎性体活性增加; 5）IL 1 β，NLRP 3的最终执行者 炎性小体激活，脊髓密度降低，谷氨酸受体离子型NMDA增加 NLRP 3炎性小体活性在SIV感染的小鼠脑中显示增加; 猕猴。基于这些发现和NLRP 3炎性小体激活的两个不同步骤，我们假设 在HIV-TAT/HIV、可卡因、抗逆转录病毒药物的背景下， 过度的镁激活和神经元损伤，这是导致神经精神疾病发病率高的原因。 使用可卡因的艾滋病毒携带者的疾病。我们将在以下两个特定目标（SA）中测试这一假设， 互补的体外和体内方法。SA 1：研究NLRP 3炎性体在Mg中的作用 在HIV-TAT/HIV、可卡因和抗逆转录病毒药物的背景下，激活和神经元损伤。我们将把这个SA分成三个 子目标。SA 1A将探索负责夸大NLRP 3炎性小体的详细机制 SA 1B将探索NLRP 3炎性小体介导的神经元激活的潜在机制。 SA 1C将研究存档的SIV中NLRP 3炎性小体和神经元损伤的状态。 感染的猕猴大脑和HIV感染的个体，无论是否使用可卡因。SA 2：探索潜力 NLRP 3炎性体抑制对体内HIV iTat小鼠中神经精神行为的治疗作用。

Abstract Although people living with HIV (PLWHIV) have comparable life-expectancy as the HIV-negative population does, their life-quality is still deeply compromised due to the prevalence of neuropsychiatric disorders including depression and anxiety. The main pathological changes in the brains of those people include aberrant microglial (Mg) activation and neuronal injuries. Drug abuse is a high comorbidity of HIV infection and abused drugs could exaggerate the existing neurologic complications in PLWHIV. The detailed mechanisms underlying such phenomenon remain much elusive. Several contributing factors have been suggested for such neurological disorders including the continued expression HIV proteins such as trans-activator of transcription (TAT), the long-term use of antiretrovirals (ARVs), and drugs of abuse. Our preliminary data demonstrated that :1) NLRP3 inflammasome signaling was involved in HIV-TAT-mediated Mg activation; 2) cocaine activated Mg through dysregulating miR-124/TLR4 axis, which could be reversed by inhibition of NLRP3 inflammasome; 3) combination ARVs used in clinical practice (tenofovir:TFV, emtricitabine:FTC, and dolutegravir:DTG) could activate Mg via lysosomal dysfunction; 4) co-exposure of Mg to three agents (TAT, cocaine, and ARVs) intriguingly resulted in increased activity of NLRP3 inflammasome in vitro; 5) IL1β, the final executor of NLRP3 inflammasome activation, decreased the spine density and increase the glutamate receptor ionotropic NMDA subunits (Grins) in vitro; and 6) increased NLRP3 inflammasome activity was shown in the brains of SIV-infected macaque. Based on these findings and two distinct steps of NLRP3 inflammasome activation, we hypothesize that exacerbated NLRP3 inflammasome activation in the context of HIV-TAT/HIV, cocaine, ARVs will lead to exaggerated Mg activation and neuronal injuries, which are responsible for the high incidence of neuropsychiatric disorders in PLWHIV with cocaine use. We will test this hypothesis in the following two specific aims (SA) using complimentary in vitro and in vivo approaches. SA1: Investigate the role of NLRP3 inflammasome in Mg activation and neuronal injuries in the context of HIV-TAT/HIV, cocaine, & ARVs. We will split this SA into three sub aims. SA1A will explore the detailed mechanisms responsible for exaggerated NLRP3 inflammasome activation in vitro; SA1B will explore the mechanisms underlying NLRP3 inflammasome-mediated neuronal injuries; and SA1C will investigate the status of NLRP3 inflammasome and neuronal injuries in archived SIV- infected macaque brains and HIV-infected individuas with or without cocaine use. SA2: Explore the potential therapeutic effects of NLRP3 inflammasome inhibition on neuropsychiatric behaviors in HIV iTat mice in vivo.