The regulation of cannabinoid receptors in microglial cells during HIV infection

HIV感染过程中小胶质细胞大麻素受体的调节

• 批准号: 8227978
• 负责人: Catalin Filipeanu
• 金额: $ 14.2万
• 依托单位: LSU HEALTH SCIENCES CENTER
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2013-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8227978
• 关键词: Accounting; Acquired Immunodeficiency Syndrome; Adrenergic Receptor; Affect; Agonist; Arrestins; Binding; Biological; Cannabinoids; Cells; Chronic; Chronic Disease; Cognitive; Cyclic AMP; Data; Dementia; Development; Disease; Disease Progression; Drug Delivery Systems; Eating; Foundations; G-Protein-Coupled Receptors; GTP-Binding Proteins; HIV; HIV Infections; HIV-1; Health Sciences; Heart failure; Highly Active Antiretroviral Therapy; Hippocampus (Brain); Human; Illicit Drugs; Immune; Immune response; Incidence; Infection; Inflammatory; Ligands; MAP Kinase Gene; Macaca mulatta; Marijuana; Mediating; Microglia; Neuraxis; Neuroglia; Neuronal Plasticity; Neurons; Outcome; Patients; Pharmaceutical Preparations; Phase; Play; Process; Proteins; Receptor Activation; Receptor Signaling; Regulation; Role; SIV; Scaffolding Protein; Signal Pathway; Signal Transduction; Symptoms; Syndrome; Testing; Tetrahydrocannabinol; Time; United States; Up-Regulation; Viral Load result; Virus Diseases; animal extract; arrestin 2; base; cannabinoid receptor; cell type; design; improved; in vivo; mortality; novel therapeutics; public health relevance; receptor; receptor expression; research study

DESCRIPTION (provided by applicant): There are over one million people in the United States infected with HIV, and due to the introduction of highly active antiretroviral therapy, HIV infection has become a chronic disease frequently co-existing with chronic use of prescribed, experimental, and illicit drugs, including marijuana. In addition to being the most potent psychoactive component of marijuana, 9-tetrahydrocannabinol (THC) has many other biological actions including modulation of the immune response, neuronal plasticity and food intake. These actions are mediated by interactions with at least three specific receptors (CB1R, CB2R and GPR55) that are expressed in the central nervous system (CNS). However, there are currently no data on the effects of HIV-infection on expression levels and signaling of cannabinoid receptors, or on the effects of cannabinoid receptor activation on the progression of HIV-infection. In our recent studies we observed that long-term THC treatment in SIV- infected rhesus macaques was found to have beneficial effects like decreasing viral load and decreasing early mortality. To test if these effects were due to changes in cannabinoid receptor expression, CB1R and CB2R protein levels were determined in hippocampal extracts. Surprisingly, SIV-infection has differential effects, CB1R levels being reduced to 42 1 8 %, whereas CB2R levels were increased with 53 1 10%. To elucidate the cellular mechanisms underlying these changes, this project will focus on the effects of HIV-1 on the cannabinoid receptors in microglial cells, as these are main immune cells at this level, playing major roles in HIV1-associated dementia (HAD), which affects 10-20 % of AIDS patients. In preliminary experiments we found that both CB1R and CB2R are expressed in human primary microglial cells, but only CB2R levels were upregulated four days post HIV-1 infection. Despite this increase, the effects of CB2R stimulation on the cAMP levels were blocked by HIV-1 infection. In contrast, CB1R activation did not change cAMP levels, but MAPK activation by this receptor was apparent only after HIV-1 infection. In addition, we found that the cellular levels of the scaffold protein 2-arrestin were upregulated after HIV-1 infection, possibly contributing to the observed changes in the cannabinoid signaling. Based on these observations we hypothesize that HIV-1 infection changes the expression levels and signaling of cannabinoid receptors in microglial cells, at least in part by altering 2-arrestin cellular levels. In the Specific aim 1 we will test if HIV-1 infection specifically increases the CB2R levels in human microglial cells, and in the Specific Aim 2 we will test if HIV-1 infection alters the CB1R- and CB2R-mediated effects on cAMP and MAPK levels through alterations in the cellular 2-arrestin levels. When completed, the results from these studies will show for the first time that HIV-1 infection modulates the expression and signaling of cannabinoid receptors in human microglial cells and these data will also provide a clearer understanding of the cellular mechanisms contributing to HIV disease progression in CNS and possibly identify new therapeutics for HIV infection by identifying new drug targets for the treatment of HAD. PUBLIC HEALTH RELEVANCE: The number of microglial cells, the main immunological cell type present in CNS, are increasing during inflammatory processes, including HIV infection, and it has been proposed that the activation of these cells contribute to HIV1-associated dementia (HAD), which has an incidence of 10-20% in AIDS patients. The function of the cannabinoid receptors, proteins which are present in these cells and recognize the main active compound from marijuana, as well as other compounds which are naturally present in the body, has not been investigated in AIDS, although there are indications that drugs interfering with the activity of the cannabinoid receptors may produce positive outcomes in the inflammatory diseases. This application will characterize the regulation of cannabinoid receptor number and function in control and HIV1 infected microglial cells, providing foundation for development of new treatments in AIDS, particularly in HAD.

描述(申请人提供)：美国有100多万人感染艾滋病毒，由于引入了高效抗逆转录病毒治疗，艾滋病毒感染已成为一种慢性病，经常与包括大麻在内的处方药物、实验药物和非法药物的长期使用并存。9-四氢大麻酚(THC)除了是大麻中最有效的精神活性成分外，还具有许多其他生物学作用，包括调节免疫反应、神经元可塑性和食物摄入量。这些作用是通过与中枢神经系统(CNS)中表达的至少三个特定受体(CB1R、CB2R和GPR55)的相互作用来介导的。然而，目前还没有关于艾滋病毒感染对大麻素受体表达水平和信号的影响，也没有关于大麻素受体激活对艾滋病毒感染进展的影响的数据。在我们最近的研究中，我们观察到，对感染SIV的恒河猴进行长期的THC治疗被发现具有减少病毒载量和降低早期死亡率等有益效果。为了验证这些效应是否源于大麻素受体表达的变化，我们测定了海马区提取物中CB1R和CB2R的蛋白水平。令人惊讶的是，SIV感染有不同的影响，CB1R水平下降到421.8%，而CB2R水平上升了53.1110%。为了阐明这些变化背后的细胞机制，本项目将重点研究HIV-1对小胶质细胞中大麻素受体的影响，因为这些细胞是这一水平的主要免疫细胞，在影响10%-20%的艾滋病患者的HIV1相关性痴呆(HAD)中发挥主要作用。在初步实验中，我们发现CB1R和CB2R在人类原代小胶质细胞中都有表达，但只有CB2R在HIV-1感染后四天表达上调。尽管有这种增加，但CB2R刺激对cAMP水平的影响被HIV-1感染所阻断。相反，CB1R的激活没有改变cAMP水平，但该受体仅在HIV-1感染后才明显激活MAPK。此外，我们发现HIV-1感染后支架蛋白2-arrestin的细胞水平上调，可能是导致观察到的大麻素信号变化的原因之一。基于这些观察，我们假设HIV-1感染改变了小胶质细胞中大麻素受体的表达水平和信号，至少部分是通过改变2-芳香素细胞水平。在特定目标1中，我们将测试HIV-1感染是否特异性地增加人小胶质细胞中CB2R水平，在特定目标2中，我们将测试HIV-1感染是否通过改变细胞内2-arrestin水平来改变CB1R和CB2R介导的对cAMP和MAPK水平的影响。当这些研究完成后，这些研究的结果将首次表明HIV-1感染调节了人类小胶质细胞中大麻素受体的表达和信号传递，这些数据还将使人们更清楚地了解导致中枢神经系统艾滋病毒疾病进展的细胞机制，并可能通过确定治疗HAD的新药物靶点来寻找治疗艾滋病毒感染的新疗法。 公共卫生相关性：小胶质细胞是中枢神经系统中存在的主要免疫细胞类型，在包括HIV感染在内的炎症过程中，小胶质细胞的数量正在增加，已有研究表明，这些细胞的激活与HIV1相关性痴呆(HAD)有关，HAD在艾滋病患者中的发病率为10%-20%。大麻素受体是存在于这些细胞中并识别来自大麻的主要活性化合物的蛋白质，以及其他天然存在于体内的化合物，尽管有迹象表明，干扰大麻素受体活性的药物可能在炎症性疾病中产生积极的结果，但在艾滋病中尚未对其功能进行研究。这一应用将表征对照和HIV1感染的小胶质细胞中大麻素受体的数量和功能的调节，为艾滋病特别是HAD的新治疗方法的开发提供基础。

项目成果

Ovarian hormones and chronic administration during adolescence modify the discriminative stimulus effects of delta-9-tetrahydrocannabinol (Δ⁹-THC) in adult female rats.

• DOI: 10.1016/j.pbb.2012.06.008
• 发表时间: 2012-09
• 期刊: PHARMACOLOGY BIOCHEMISTRY AND BEHAVIOR
• 影响因子: 3.6
• 作者: Winsauer, Peter J.;Filipeanu, Catalin M.;Bailey, Evangeline M.;Hulst, Jerielle L.;Sutton, Jessie L.
• 通讯作者: Sutton, Jessie L.