A new pathway of spinal neurons in neuropathic pain induced by HIV with opioid

项目总结： 为回应RFA-MH-18-610-“FOA邀请研究拨款申请，以破译路径和 HIV-1导致中枢神经系统(CNS)功能障碍的机制“，我们重点关注 研究兴趣领域：“NIDA感兴趣的是了解潜在的机制 滥用药物和艾滋病毒感染相互作用，通过改变神经元回路来损害中枢神经系统功能， 神经感受器。美国目前存在阿片类药物过度使用的流行病。尽管出现了 有效的抗逆转录病毒治疗，HIV-神经病理性疼痛(HIV-NP)是一种常见的神经系统并发症，如 患者享有更长的寿命。长期滥用鸦片类药物进一步加剧了这一严重的健康问题 通常见于HIV+的个体，进而导致神经缺陷的严重程度增加。艾滋病毒与慢性 使用/滥用吗啡可增加细胞因子的产生，导致神经炎症加剧。与HIV相关的 神经毒性物质直接或通过激活神经胶质细胞间接引起神经元损伤 释放神经毒性因子(如肿瘤坏死因子α)。我们的初步数据显示，反复接触艾滋病毒衣物 蛋白gp120与吗啡一起降低NAD+依赖的神经元sirtuin(SIRT)3的表达 调节脊髓背角线粒体解毒的脱乙酰酶，并增加 神经元线粒体中的超氧化物。然而，造成损失的确切上游路径 神经元SIRT3在SCDHgp120和吗啡相互作用中的作用尚不清楚。最近的研究表明， 表观遗传阅读器Brd4是溴域和端外(BET)家族的一种蛋白质，它调节 转录因子c-Myc的表达。Zust同源基因2(EZH2)的表观遗传学编写者和增强子 通过组蛋白甲基化抑制基因表达(例如，H3K27me3)。BRD4正向调节EZH2 通过上调c-Myc转录。在这项提议中，我们将检验神经元TNFRI- --Brd4-c-Myc-EZH2表观遗传途径介导脊髓中抗氧化SIRT3系统的丧失 脊髓背角在HIV/慢性阿片类药物相关神经病理性疼痛中的作用。我们将把分子遗传学和表观遗传学结合起来 方法，基因敲除(KO)小鼠，条件基因敲除(CKO)小鼠，神经元选择性病毒载体， 线粒体超氧化物成像和分子药理学分析都在体内证明了新的 机械装置。基于神经元信号转导途径，这些研究的结果将提供 艾滋病病毒与阿片类药物相互作用对神经病理性疼痛治疗的启示。

Project Summary: In response to the RFA-MH-18-610---“the FOA invites research grant applications to decipher pathways and mechanisms responsible for HIV-1 induced central nervous system (CNS) dysfunction”, we focused on the Areas of Research Interest: “NIDA is interested in understanding the underlying mechanisms whereby drugs of abuse and HIV infection interact to impair CNS functions mediated through altered neuronal circuits, neuronal receptors.” The United States currently has an opioid overuse epidemic. Despite the advent of effective anti-retroviral therapy, HIV-neuropathic pain (HIV-NP) is a common neurological complication as patients enjoy longer life spans. This serious health issue is further exacerbated by chronic abuse of opiates often seen in HIV+ individuals, leading, in turn, to increased severity of neurological deficits. HIV and chronic morphine use/abuse can increase cytokine production resulting in enhanced neuroinflammation. HIV-related neuron damage is induced either directly by neurotoxic substances or indirectly by activating glial cells releasing neurotoxic factors (such as, TNFα). Our preliminary data shows that repeated exposure of HIV coat protein gp120 with morphine decreased neuronal sirtuin (SIRT) 3 expression, an NAD+-dependent deacetylase that regulates mitochondrial detoxification in the spinal cord dorsal horn (SCDH), and increased in neuronal mitochondrial superoxide. However, the exact upstream pathways responsible for the loss of neuronal SIRT3 in the SCDH in the interaction of gp120 and morphine is not clear. Recent studies show that the epigenetic reader Brd4, one of bromodomain and extraterminal (BET) family of proteins modulates, the expression of transcriptional factor c-Myc. The epigenetic writer, enhancer of zeste homology 2 (EZH2) suppresses gene expression via histone methylation (e.g., H3K27me3). Brd4 positively regulates EZH2 transcription through upregulation of c-Myc. In this proposal we will test the hypothesis that neuronal TNFRI- --Brd4---c-Myc---EZH2 epigenetic pathway mediates the loss of anti-oxidative SIRT3 system in the spinal cord dorsal horn in HIV/chronic opioid-related neuropathic pain. We will combine molecular and epigenetic approach, knockout (KO) mice, conditional knockout (cKO) mice, neuron-selective viral vectors, mitochondrial superoxide imaging, and molecular-pharmacological assays all in vivo to prove novel mechanisms. Based on the neuronal signal transduction pathway, the results of these studies will provide insights on the treatment of neuropathic pain of the interaction of HIV and opioids.