Opioid Impact on Trim37-Induced Restriction of HIV

阿片类药物对 Trim37 诱导的 HIV 限制的影响

• 批准号: 8525374
• 负责人: Lauren Brittany Beach
• 金额: $ 0.42万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-30 至 2013-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8525374
• 关键词: Acquired Immunodeficiency Syndrome; Affect; Anti-Retroviral Agents; Binding; Biological Assay; California; Cattle; Cell Line; Cells; Cercopithecus pygerythrus; Co-Immunoprecipitations; Cytokine Receptors; DNA; DNA biosynthesis; Dendritic Cells; Development; Drug Addiction; Drug user; Enhancers; Equus caballus; Family member; Felis catus; Future; Genetic Transcription; Glioblastoma; HIV; HIV-1; Health; Human; Human Cell Line; Immune; Immune response; Immune system; Immunofluorescence Immunologic; Infection; Interferon Type I; Interferons; Laboratories; Leukocytes; Ligands; Light; Lipopolysaccharides; Luciferases; Macaca mulatta; Messenger RNA; Monitor; Morphine; Mus; NF-kappa B; NFAB complex; Night Monkey; Nuclear; Opioid; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Process; Proteins; Publishing; Receptor Signaling; Relative (related person); Reporter; Research; Reverse Transcription; Signal Transduction; TNF Receptor-Associated Factors; TRAF2 gene; TRAF6 gene; TRIM Motif; Testing; Tissue Sample; Tumor Necrosis Factor Receptor; Tumor Necrosis Factor-alpha; United States; Virus; Virus Diseases; Virus Replication; Withdrawal; base; cell type; cytokine; expression vector; human TNF protein; improved; in vivo; insight; knock-down; mu opioid receptors; novel; particle; receptor; receptor expression; research study; toll-like receptor 4; transcription factor; viral DNA

This application proposes to investigate how Trim37, as a hypothesized component of the human innate immune system, influences HIV-1 replication dynamics in the presence or absence of opiods. Published research from several independent laboratories has revealed that opiods increase HIV replication ex vivo in human immune cells and in vivo in mice and rhesus macaques. It has been hypothesized that the mechanism for the observed increases in HIV-1 replication is due to opioids' well-documented ability to modulate cytokine and cytokine receptor signaling and expression. One recent study with murine dendritic cells has shown that the mu-opioid receptor, which is the receptor responsible for morphine's drug-induced effects, can be induced by the toll-like receptor 4 ligand, lipopolysaccharide (LPS). Unpublished research in the Mansky lab has demonstrated that Trim 37 is packaged into HIV-1 particles and decreases virus infectivity by reducing viral DNA synthesis. It has been previously observed that the expression of Trim37 results in the inability of TRAF proteins 2 and 6 to stimulate NF-KB binding to DNA. Notably, NF-KB is one of the major human transcription factors that transcribes HIV-1; tumor necrosis factor alpha (TNFalpha) and LPS both stimulate NF-KB's binding to DNA. This proposal seeks to determine whether various mammalian Trim37 proteins can also restrict HIV-1 infectivity in cell lines and primary cells. Both single-cycle replication and spreading HIV-1 infection assays will be performed in the presence of varying amounts of Trim37 to determine each Trim37 protein's antiretroviral effect. Due to the observations that both Trim37 and opioids can modulate NF-KB signaling, experiments will also be conducted in the presence of opioids. Additionally, experiments to determine whether Trim37's expression or localization changes in the presence of opioids, interferons I, II, and III, and TNFalpha will also be done. The determination of whether opioids alter the ability of Trim 37 to restrict HIV replication could help yield insight into the restriction mechanisms involved. Given that many of the 33.4 million HIV-1 infected people worldwide use opioids, these studies have the potential to improve global human health among HIV-1 infected opioid users.

该应用程序旨在研究Trim37作为人类先天免疫系统的假设成分，在阿片类药物存在或不存在的情况下如何影响HIV-1复制动力学。几个独立实验室发表的研究表明，阿片类药物增加了人类免疫细胞体内和小鼠和恒河猴体内的艾滋病毒复制。据推测，观察到的HIV-1复制增加的机制是由于阿片类药物调节细胞因子和细胞因子受体信号传导和表达的能力。最近对小鼠树突状细胞的一项研究表明，负责吗啡药物诱导作用的受体mu-阿片受体可以被toll样受体4配体脂多糖（LPS）诱导。曼斯基实验室未发表的研究表明，Trim 37被包装到HIV-1颗粒中，通过减少病毒DNA的合成来降低病毒的传染性。先前已经观察到，Trim37的表达导致TRAF蛋白2和6无法刺激NF-KB与DNA的结合。值得注意的是，NF-KB是转录HIV-1的主要人类转录因子之一；肿瘤坏死因子α (TNFalpha)和LPS都刺激NF-KB与DNA的结合。本研究旨在确定各种哺乳动物Trim37蛋白是否也能限制细胞系和原代细胞中的HIV-1传染性。单周期复制和传播HIV-1感染试验将在存在不同量的Trim37的情况下进行，以确定每种Trim37蛋白的抗逆转录病毒作用。由于观察到Trim37和阿片样物质都可以调节NF-KB信号，实验也将在阿片样物质存在的情况下进行。此外，我们还将进行实验，以确定Trim37在阿片样物质、干扰素I、II、III和TNFalpha存在下的表达或定位是否会发生变化。确定阿片类药物是否会改变Trim 37限制HIV复制的能力，有助于深入了解所涉及的限制机制。鉴于全世界3340万艾滋病毒-1感染者中有许多人使用阿片类药物，这些研究有可能改善艾滋病毒-1感染阿片类药物使用者的全球人类健康。