Effects of Exosomes Derived from HIV-1 Infected Cells on Viral Spread

HIV-1感染细胞衍生的外泌体对病毒传播的影响

• 批准号: 8874765
• 负责人: Gavin Sampey
• 金额: $ 0.98万
• 依托单位: GEORGE MASON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-25 至 2015-12-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8874765
• 关键词: Acquired Immunodeficiency Syndrome; Animals; Astrocytes; Binding; Biogenesis; Biological Assay; Brain; Canada; Cell Line; Cells; Complex; Control Groups; DNA; Data; Diagnosis; Distal; Environment; Epidemic; Exposure to; Gene Expression; Goals; HIV; HIV Infections; HIV-1; HIV-associated neurocognitive disorder; Highly Active Antiretroviral Therapy; Human; In Vitro; Indinavir; Individual; Infection; Knowledge; Life; Life Cycle Stages; Lipids; Lytic; Maintenance; Messenger RNA; MicroRNAs; Microglia; Molecular; Mus; NFAB complex; Outcome Study; Pathogenicity; Patients; Phenotype; Play; Predisposition; Production; Proviruses; RNA; RNA Interference Pathway; Regulation; Reporting; Response Elements; Role; Serum; Signal Transduction; Structure of thyroid parafollicular cell; T-Lymphocyte; Tenofovir; Testing; Transduction Gene; Validation; Viral; Viral Pathogenesis; Virus; Virus Diseases; cytokine; ds-DNA; emtricitabine; in vivo; inhibitor/antagonist; intercellular communication; killings; macrophage; manumycin; monocyte; mouse model; mutant; pathogen; public health relevance; research study; response; stem; virus host interaction

DESCRIPTION (provided by applicant): As of 2011, the UNAIDS report on the global AIDS epidemic indicated that 34 million people are living with HIV. Regionally, the number of infected individuals in the USA and Canada is 1.4 million with 51,000 new cases being documented every year. Furthermore, AIDS-related causes claim the lives of nearly 20,000 people in the USA annually and HIV-associated neurocognitive disorders are diagnosed in up to 47 percent of patients despite the use of highly active antiretroviral therapy. Recently, the concept of exosomes acting as vehicles of intercellular communication has opened up new avenues of study regarding host-pathogen interactions and viral spread. Another emerging field regarding viral infections is that of the RNA interference pathway which generates miRNAs that guide effector complexes to complimentary sequences in mRNA or DNA, thereby suppressing gene expression. Moreover, our lab, as well as others, has identified miRNAs derived from the TAR element of integrated provirus in long-term infections. While the concept of HIV-1 viral miRNAs remains controversial, labs concluding that HIV-1 doesn't encode miRNAs have examined short-term infections as opposed to the more biologically relevant long-term infections. Furthermore, our initial data shows viral miRNAs derived from HIV-1 TAR are packaged and secreted from infected cells through exosomes. Our long term goal is to understand the role played by exosomes from HIV-1 infected cells in regulating host-virus interactions. The primary objective of the current proposal is to elucidate the mechanisms by which HIV altered exosomes effect uninfected cells of CNS origins, and assist in viral spread. We hypothesize that viral miRNAs within exosomes of infected cells alter recipient cells by impacting regulation signal transduction and gene expression, as well as viral spread. The rationale for our hypothesis includes our data showing viral miRNAs in culture supernatants and patient sera suggesting the miRNAs are in a protected environment. Moreover, we've shown that exosomes from infected cells can exert functional influences on naive bystander cells and make them more susceptible to HIV infection. However, our current understanding of the functionality of HIV-altered exosomes is limited, resulting in a gap of knowledge regarding exosomal mechanisms that contribute to viral pathogenesis in the CNS. To test our hypothesis we propose the following aims: 1) Examine the altered composition and mechanisms of exosome manipulation in infected primary cells, and validate the functionality of exosomes on recipient cells of CNS origins 2) Test the effects of exosomes on HIV-1 replication in human MDMs within the brain using humanized animals. The expected outcomes of these studies includes elucidation of altered exosome composition and viral miRNA influences on signal transduction, gene expression and increased viral susceptibility of naive recipient cells of the CNS. Our in vivo experiments, in combination with validation of decreased exosome biogenesis following exosome inhibitor treatment, will establish a strong correlation between circulating exosomes and infection-associated phenotypes in the CNS.

描述（由申请人提供）：截至2011年，《联合国艾滋病规则》报告的全球艾滋病流行报告表明，有3400万人患有艾滋病毒。在区域上，美国和加拿大受感染的人数为140万，每年记录51,000例新病例。此外，与AIDS相关的原因声称，尽管使用了高度活跃的抗逆转录病毒疗法，但在美国近47％的患者中，每年在美国近20,000人的生活和与HIV相关的神经认知疾病的生命被诊断出来。最近，充当细胞间交流车辆的外泌体的概念开辟了有关宿主病原体相互作用和病毒蔓延的新研究途径。关于病毒感染的另一个新兴领域是RNA干扰途径的miRNA，该途径引导效应子复合物以mRNA或DNA中的免费序列，从而抑制基因表达。此外，我们的实验室以及其他实验室已经确定了长期感染中综合病毒的焦油元素得出的miRNA。尽管HIV-1病毒miRNA的概念仍然引起争议，但实验室得出结论，HIV-1没有编码miRNA检查了短期感染，而不是生物学上相关的长期感染。此外，我们的初始数据表明，源自HIV-1焦油的病毒miRNA被包装并通过外泌体分泌从感染的细胞中分泌。我们的长期目标是了解来自HIV-1感染细胞的外泌体在调节宿主病毒相互作用中所起的作用。当前建议的主要目的是阐明艾滋病毒改变外泌体对中枢神经系统起源的未感染细胞的机制，并有助于病毒扩散。我们假设感染细胞外泌体内的病毒miRNA通过影响调节信号转导和基因表达以及病毒扩散而改变受体细胞。我们的假设的基本原理包括我们的数据显示培养上清液中的病毒miRNA和患者血清中，表明miRNA处于受保护的环境中。此外，我们已经表明，来自感染细胞的外泌体可以对幼稚的旁观者细胞产生功能影响，并使它们更容易受到HIV感染。但是，我们目前对改变HIV的外泌体功能的理解受到限制，从而导致有关在CNS中导致病毒发病机理的外泌体机制的知识差距。为了检验我们的假设，我们提出了以下目的：1）检查感染原代细胞中外泌体操纵的组成和机制的改变，并验证外泌体对CNS起源受体细胞的功能2）测试外泌体对使用人类动物使用人类动物中人类MDMS中HIV-1复制的外壳影响。这些研究的预期结果包括阐明外泌体组成改变和病毒miRNA对信号转导，基因表达的影响以及CNS天真受体细胞的病毒易感性增加。我们的体内实验，结合验证外泌体抑制剂治疗后的外泌体生物发生的验证，将在中枢神经系统中循环外泌体和与感染相关的表型之间建立很强的相关性。