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

项目概要： 为了回应RFA-MH-18-610-“FOA邀请研究资助申请破译途径， HIV-1诱导的中枢神经系统（CNS）功能障碍的机制”，我们专注于 研究兴趣领域：“NIDA有兴趣了解潜在的机制， 药物滥用和HIV感染相互作用，通过改变神经元回路介导损害CNS功能， 神经受体。”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，艾滋病毒神经性疼痛（HIV-NP）是一种常见的神经系统并发症， 患者的寿命更长。这一严重的健康问题因长期滥用阿片类药物而进一步恶化 这通常见于HIV+个体，进而导致神经功能缺损的严重程度增加。艾滋病毒和慢性 吗啡的使用/滥用可增加细胞因子的产生，导致神经炎症的增强。hiv相关 神经元损伤可由神经毒性物质直接或通过激活神经胶质细胞间接诱导 释放神经毒性因子（如TNFα）。我们的初步数据显示，反复接触艾滋病毒外壳 蛋白gp 120与吗啡降低神经元sirtuin（SIRT）3的表达，这是一种NAD+依赖性的 在脊髓背角（SCDH）中调节线粒体解毒的脱乙酰酶， 神经元线粒体超氧化物。然而，负责损失的确切上游途径， 神经元SIRT 3在SCDH中与gp 120和吗啡的相互作用尚不清楚。近年来研究表明 表观遗传阅读器Brd 4，溴结构域和末端外（BET）家族蛋白质之一，调节 转录因子c-Myc的表达。表观遗传学作者，zeste同源性增强子2（EZH 2） 通过组蛋白甲基化抑制基因表达（例如，H3K27me3）。Brd 4正向调节EZH 2 通过上调c-Myc转录。在这个提议中，我们将测试神经元TNFRI- --Brd 4-c-Myc-EZH 2表观遗传通路介导脊髓抗氧化SIRT 3系统的丧失 脊髓背角在HIV/慢性阿片类药物相关神经性疼痛中的作用我们将联合收割机结合分子和表观遗传学 方法，敲除（KO）小鼠，条件性敲除（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试验，所有这些都在体内证明了新的 机制等基于神经元信号转导通路，这些研究结果将提供 对艾滋病毒和阿片类药物相互作用的神经性疼痛治疗的见解。

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.