Circulating Cellular and Extracellular Noncoding RNAs in HIV-1 Elite Suppression

描述(申请人提供)：在没有抗逆转录病毒治疗(ART)的情况下，HIV-1感染通常伴随着CD4T细胞计数下降，病毒载量上升，最终 进展为艾滋病和死亡。通过不完全了解的机制，少数人将血浆病毒载量长期维持在临床检测极限(50拷贝/毫升)以下，即使没有抗逆转录病毒治疗。更好地了解这些精英抑制子或控制器如何抑制病毒复制可能有助于开发有效的疫苗和新的治疗方法。一类小的调节分子-被称为microRNAs(MiRNAs)的短核酸-提供了最近才发现的转录后调节层。MiRNAs也以一种特定的方式从细胞中输出，并且在血液中足够稳定，可以建立细胞间的通信。尽管人们对这些分子产生了浓厚的兴趣，并在多个研究领域取得了丰硕的成果，但关于细胞或细胞外miRNA参与HIV-1控制和进展的信息相对较少，几乎没有关于ES中miRNAs的信息。我们最近发表了第一个关于HIV疾病灵长类动物模型中感染和CNS疾病的血浆miRNA生物标志物的报告，以及第一个在ES人群中进行的miRNAs研究，在这些研究中，我们发现PBMC miRNAs聚集了来自健康对照组的慢性进展者，同时暗示ES现象可能有多种机制。这些结果，结合我们小组和其他人在ES、慢性进展性疾病和对照的T细胞亚群的定性和定量差异的发现，促使我们提出这个项目。我们的目标是描述miRNA与精英抑制、HIV-1感染的正常进展以及对ART的反应之间的关系。我们假设：1)存在ES、HIV进展和对ART反应的特定细胞和血浆miRNA生物标志物；2)miRNAs参与有助于病毒控制和使ES细胞抵抗HIV介导的细胞死亡的途径；3)细胞外miRNAs不均匀地分布在大小和功能不同的血浆颗粒中，表明特定的释放和参与免疫协调网络。为了验证这些假说，我们将从特征良好的ES队列、接受ART治疗和ART治疗的慢性进展者和健康对照中收集样本。在目标1中，我们将分析参与精英抑制和HIV复制的六种特定免疫细胞类型中miRNAs的水平：CD4和CD8 NA、效应器记忆细胞和中央记忆细胞。这一目标还将检查miRNA图谱与艾滋病毒疾病的经典参数之间的关系。在目标2中，我们将表征在囊泡、脂蛋白颗粒和蛋白质复合体中循环的胞外血浆miRNAs，并确定特定的血浆miRNAs是否与来自CD4或CD8T细胞的血浆纳米颗粒的丰度和/或大小有关。该项目的发现将与基础miRNA科学相关，并将通过发现生物标记物和确定基于ES现象的miRNA疗法的可行性来推动HIV-1临床科学的发展。

DESCRIPTION (provided by applicant): In the absence of antiretroviral therapy (ART), HIV-1 infection is typically followed by declining CD4+ T-cell counts, elevated viral loads, and eventual progression to AIDS and death. By means of incompletely understood mechanisms, a minority of individuals maintain plasma viral loads under the clinical limit of detection (50 copies/mL) ove the long term, even without ART. A better understanding how these elite suppressors or controllers (ES) restrain virus replication may aid in developing effective vaccines and new treatments. A class of small regulatory molecules-short nucleic acids known as microRNAs (miRNAs)- provide an only recently discovered layer of post-transcriptional regulation. miRNAs are also exported from the cell in a specific fashion and are sufficiently stable in blood to establish cell-to-cell communication. Despite intense interest in these molecules and productive findings in multiple fields of study, there is relatively little information on cellular or extracelular miRNA involvement in HIV-1 control and progression, and virtually none on miRNAs in ES. We recently published the first report of plasma miRNA biomarkers of infection and CNS disease in a primate model of HIV disease, as well as the first study of miRNAs in the ES population, in which we found that PBMC miRNAs cluster chronic progressors from healthy controls while suggesting that more than one mechanism may underlie the ES phenomenon. These results, combined with the findings by our group and others of qualitative and quantitative differences in T-cell subsets of ES, chronic progressors, and controls, have prompted us to propose this project. We aim to characterize miRNA associations with elite suppression, normal progression of HIV-1 infection, and the response to ART. We hypothesize that: 1) there exist specific cellular and plasma miRNA biomarkers of ES, HIV progression, and response to ART; 2) miRNAs are involved in pathways that contribute virus control and render ES cells resistant to HIV-mediated cell death; and 3) extracellular miRNAs are nonuniformly distributed among size- and functionally distinct plasma particles, indicating specific release and involvement in immune coordination networks. To test these hypotheses, we will collect samples from a well-characterized ES cohort, ART-treated and ART-na¿ve chronic progressors, and healthy controls. In Aim 1, we will analyze levels of miRNAs in six specific immune cell types that are involved in elite suppression and HIV replication: CD4+ and CD8+ na¿ve, effector memory, and central memory cells. This aim will also examine the relationship of miRNA profiles and classical parameters of HIV disease. In Aim 2, we will characterize extracellular plasma miRNAs, which circulate in vesicles, lipoprotein particles, and protein complexes, and determine whether specific plasma miRNAs are associated with abundance and/or size of plasma nanoparticles that derive from CD4+ or CD8+ T-cells. The findings of this project will have relevance for basic miRNA science and will advance HIV-1 clinical science through biomarker discovery and determining the feasibility of miRNA-based therapies based on the ES phenomenon.