Immunomodulation of nicotine in HIV-1Tg rat brain

项目总结： 作为烟草烟雾的主要活性成分之一，尼古丁主要作用于烟碱。 乙酰胆碱受体(NAChRs)，表达在神经元、小胶质细胞和其他中央细胞上。 神经系统(CNS)。尼古丁激活nAChRs具有多种潜在的治疗免疫益处， 包括调节炎症反应、维持免疫动态平衡和调节 促炎和抗炎因子的表达。然而，吸烟者感染艾滋病毒的几率更高。 进展与行为和认知并发症，男性和女性之间存在差异 吸烟者。已有研究表明，通过吸烟使HIV患者长期接触尼古丁 可能减少nAChRs的表达并使其失敏，从而抑制尼古丁的能力 来调节免疫反应。在过去的几年里，我们进行了一系列的行为和 使用啮齿动物HIV-1转基因(HIV-1TG)大鼠模型进行的分子研究，以研究各种医学问题， 包括免疫，与接受联合抗逆转录病毒治疗(CART)的艾滋病毒患者相关。我们用了 在HIV-1存在的情况下确定尼古丁对脑功能影响的HIV转基因大鼠模型 蛋白质，并发现长期服用尼古丁可以恢复许多基因的表达，这些基因由 HIV-1病毒蛋白在信号通路中参与免疫和其他神经系统。此外，我们 发现多种nAChR亚基，特别是α6，β3和β4，以及免疫相关基因，如 作为干扰素调节因子7(Irf7)和白细胞介素1α(IL-1α)，在HIV-1TG大鼠与 F344控制。我们的发现表明，HIV-1蛋白对中枢神经系统免疫和nAChR都有很大的影响 在某些免疫相关和nAChR介导的信号中发挥作用并改变对尼古丁的反应性 小路。基于这些发现，我们假设尼古丁和尼古丁之间存在显著的相互作用。 HIV-1阳性个体大脑中影响尼古丁免疫反应的HIV-1蛋白。为了测试这一点 假设，我们提出了以下三个目标。目标1是确定尼古丁和尼古丁的交互作用 脑内小胶质细胞和巨噬细胞上的HIV病毒蛋白以及血液和血液中细胞因子的产生 HIV-1TG大鼠的大脑。目的2是比较尼古丁和HIV-1病毒蛋白对 用rna-seq方法分析雄性和雌性HIV-1TG大鼠三个脑区的基因表达。目标3是 描述调节尼古丁和HIV之间相互作用的特定基因和途径- 1病毒蛋白使用各种分子技术，包括CRISPR基因编辑。建议进行的研究 代表了下一代测序技术与传统测序技术相结合的首次应用 研究长期使用尼古丁对中枢神经系统免疫调节作用的分子方法 HIV-1病毒蛋白的存在。通过将基因组和分子方法结合起来，我们的研究将是 全面，没有主观偏见，保证数据的价值，为新的发展 治疗吸食尼古丁的HIV-1患者的策略。

Project Summary: As one of the key active ingredients of tobacco smoke, nicotine exerts its actions primarily on the nicotinic acetylcholine receptors (nAChRs), which are expressed on neurons, microglia, and other cells in the central nervous system (CNS). Activation of nAChRs by nicotine has various potential therapeutic immune benefits, including modulation of inflammatory responses, maintenance of immune homeostasis, and regulation of expression of pro- and anti-inflammatory factors. However, smokers have a higher rate of HIV disease progression and behavioral and cognitive complications, with differences seen between male and female smokers. It has been suggested that prolonged exposure to nicotine in HIV patients through cigarette smoking might decrease the expression and desensitize the activation of nAChRs, which could dampen nicotine's ability to mediate immune responses. During the past several years, we have conducted a series of behavioral and molecular studies using a rodent HIV-1 transgenic (HIV-1Tg) rat model to study various medical issues, including immunity, associated with HIV patients receiving combined anti-retroviral therapy (cART). We used this HIV-transgenic rat model to determine the effects of nicotine on brain function in the presence of HIV-1 proteins and found that chronic treatment with nicotine can restore the expression of many genes altered by the HIV-1 viral proteins in signaling pathways involved in immunity and other neuronal systems. In addition, we found that expression of various nAChR subunits, especially α6, β3, and β4, and immune-related genes, such as interferon regulatory factor 7 (IRF7) and interleukin-1α (IL-1α), differ greatly in HIV-1Tg rats compared with F344 controls. Our findings indicate that HIV-1 proteins greatly impact CNS immunity as well as nAChR function and alter the responsiveness to nicotine in certain immune-related and nAChR-mediated signaling pathways. Based on these findings, we hypothesize that there are significant interactions between nicotine and HIV-1 proteins that affect the immune response to nicotine in the brain of HIV-1-positive individuals. To test this hypothesis, we propose the following three aims. Aim 1 is to determine the interactive effects of nicotine and HIV viral proteins on microglia and macrophages in the brain as well as cytokine production in the blood and brain of the HIV-1Tg rat. Aim 2 is to compare the interactive effects of nicotine and HIV-1 viral proteins on gene expression in three brain regions of male and female HIV-1Tg rats using RNA-seq analysis. Aim 3 is to characterize specific genes and pathways that mediate the effects of the interaction between nicotine and HIV- 1 viral proteins using various molecular techniques, including CRISPR gene editing. The proposed studies represent the first application of next-generation sequencing technique in combination with conventional molecular approaches to investigate the modulatory effects of chronic use of nicotine on CNS immunity in the presence of HIV-1 viral proteins. By combining both genomic and molecular approaches, our studies will be comprehensive and free of subjective bias and assure the value of the data for the development of new strategies for treating HIV-1 patients who use nicotine.