Immunomodulation of nicotine in HIV-1Tg rat brain

项目摘要： 作为烟草烟雾中的主要活性成分之一，尼古丁主要对尼古丁产生作用。 乙酰胆碱受体（nAChRs），其在中枢神经系统中的神经元、小胶质细胞和其他细胞上表达。 神经系统（CNS）。尼古丁激活nAChR具有多种潜在的治疗免疫益处， 包括炎症反应的调节、免疫稳态的维持和 促炎因子和抗炎因子的表达。然而，吸烟者感染艾滋病毒的几率更高 进展以及行为和认知并发症，男性和女性之间存在差异 吸烟者。有研究表明，艾滋病患者通过吸烟长期接触尼古丁 可能会减少nAChRs的表达并使其活化脱敏，这可能会抑制尼古丁的能力， 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-1 Tg）大鼠模型进行分子研究，以研究各种医学问题， 包括免疫力，与接受联合抗逆转录病毒疗法（cART）的艾滋病毒患者有关。我们使用 该HIV转基因大鼠模型用于确定尼古丁在HIV-1存在下对脑功能的影响 研究人员发现，尼古丁的长期治疗可以恢复许多基因的表达，这些基因被尼古丁改变了。 HIV-1病毒蛋白参与免疫和其他神经系统的信号通路。另外我们 发现各种nAChR亚单位，特别是α6，β3和β4，以及免疫相关基因， 如干扰素调节因子7（IRF 7）和白细胞介素-1 α（IL-1α），在HIV-1 Tg大鼠中与 F344控制。我们的研究结果表明，HIV-1蛋白极大地影响中枢神经系统免疫以及nAChR 在某些免疫相关和nAChR介导的信号传导中起作用并改变对尼古丁的反应性 途径。基于这些发现，我们假设尼古丁和 HIV-1蛋白影响HIV-1阳性个体大脑中对尼古丁的免疫反应。为了验证这一 假设，我们提出以下三个目标。目的1是确定尼古丁和 HIV病毒蛋白在大脑中的小胶质细胞和巨噬细胞上，以及血液中的细胞因子产生， HIV-1 Tg大鼠的大脑。目的2是比较尼古丁和HIV-1病毒蛋白的相互作用， 使用RNA-seq分析在雄性和雌性HIV-1 Tg大鼠的三个脑区域中的基因表达。目标3是 表征介导尼古丁和HIV之间相互作用效应的特定基因和途径- 1病毒蛋白使用各种分子技术，包括CRISPR基因编辑。拟议的研究 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。 研究慢性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。通过结合基因组和分子方法，我们的研究将是 全面和无主观偏见，并确保数据的价值，为发展新的 治疗使用尼古丁的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.