Extracellular Vesicle and Extracellular RNA Biomarkers of HIV-1 Central Nervous System Pathogenesis and Cigarette Use

摘要 艾滋病毒相关的神经认知障碍(手)和其他神经疾病预计会影响 大多数艾滋病毒感染者在一生中的某个时候。尽管有效，这些障碍还是会发生。 通过抗逆转录病毒疗法(ART)进行病毒学抑制。然而，几乎没有建立起生物标记。回顾说 为艾滋病开发的血浆生物标志物(病毒载量和CD4T细胞计数)在发展过程中是必不可少的 对于艾滋病的治疗范例，加强对神经系统疾病生物标记物的研究是必要的。 当物质使用障碍(SODS)与物质使用障碍(SOD)并存时，改进的生物标志物可能特别重要 艾滋病。在艾滋病病毒感染者中，吸烟的比例是普通人群的几倍。 并导致炎症和衰老过程，这些过程加剧了 多个器官系统。有希望的生物标记物包括稳定的胞外RNA(ExRNAs)， 其中一部分被包装成细胞外小泡(EV)，可以使脑等组织 在容易接触到的生物体液中发现。这些实体作为神经学的标志物受到越来越多的关注 疾病，因为它们在神经退行性变中起作用，(对于EVS)因为它们可以追溯到一个细胞或 起源的组织。有两类exRNA特别吸引人，因为它们具有非凡的稳定性： MicroRNAs(MiRNAs)和CircRNAs(CircRNA)。MiRNAs已经在HIV疾病和肥皂症中进行了研究， 但不是在脑组织的EVS中，而且对HIV-1致病机制或SODS中的CircRNAs几乎一无所知。在……里面 在这个项目中，我们将开发HIV-1致病机制和香烟使用的生物标志物目录，并确定如何 香烟的使用可能会使对生物标志物测试的解释复杂化。存档的样本来自一个特征良好的 将对动物模型和人类队列进行测试，以确定和验证候选生物标志物。我们的专业知识是 在HIV-1疾病和动物模型中；神经系统疾病的生物标记物；EV技术和生物学；以及 非编码RNA。我们将首先使用一种新的组织EV分离技术来寻找候选分子 EV和exRNA在脑组织中的特征(目标1)，然后调查EV和exRNA在 匹配，纵向收集脑脊液和血浆(目标2)。重要的是，Aim 3调查了 平行的人体样本。该项目将以严谨和可重现的方式解决目前文献中的空白 通过排除EV和exRNA分析的常见伪迹并通过尖端确认 孤立电动汽车的身份。总体而言，我们假设脑源性电动汽车及其最稳定的exRNA Cargo，miRNAs和CircRNAs，作为艾滋病毒相关神经疾病的生物标志物。因为有些人 标志物也将与吸烟有关，烟草使用将被调查是否可能产生干扰 对神经系统疾病生物标志物测量的影响。

ABSTRACT The HIV-associated neurocognitive disorders (HAND) and other neurologic disorders are predicted to affect a majority of HIV-infected individuals at some time during the lifespan. These disorders occur despite effective virologic suppression by antiretroviral therapy (ART). Yet few established biological markers. Recalling that plasma biomarkers developed for AIDS (viral load and CD4+ T cell count) were essential in developing treatment paradigms for AIDS, an intensified research focus on biomarkers of neurologic disease is warranted. Improved biomarkers may be particularly important when substance use disorders (SUDs) are comorbid with HIV disease. Among HIV+ individuals, cigarette smoking is several times more prevalent than in the general population and contributes to inflammation and senescence processes that exacerbate pathogenesis in multiple organ systems. Promising classes of biomarkers include stable extracellular RNAs (exRNAs), a portion of which are packaged into extracellular vesicles (EVs) that can leave tissue such as brain to be found in easily accessed biological fluids. These entities receive increasing attention as markers of neurologic disease because of roles in neurodegeneration and (for EVs) because they can be traced back to a cell or tissue of origin. Two categories of exRNA are particularly attractive because of their extraordinary stability: microRNAs (miRNAs) and circular RNAs (circRNAs). miRNAs have been studied in HIV disease and SUDs, but not in EVs of brain tissue, and almost nothing is known about circRNAs in HIV-1 pathogenesis or SUDs. In this project, we will develop a catalog of biomarkers of HIV-1 pathogenesis and cigarette use and identify how cigarette use may complicate interpretation of biomarker tests. Archived samples from a well characterized animal model and human cohorts will be tested to identify and validate candidate biomarkers. Our expertise is in HIV-1 disease and animal models; biomarkers of neurologic disease; EV techniques and biology; and noncoding RNAs. We will first use a novel tissue EV isolation technique to seek candidate molecular signatures of EVs and exRNA in brain tissue (Aim 1), and then investigate EV and exRNA signatures in matched, longitudinally collected cerebrospinal fluid and blood plasma (Aim 2). Importantly, Aim 3 investigates human samples in parallel. the project will address current gaps in the literature in rigorous and reproducible fashion by ruling out common artifacts of EV and exRNA analysis and through cutting edge confirmation of the identity of isolated EVs. Overall, we hypothesize that brain-derived EVs and their most stable exRNA cargo, miRNAs and circRNAs, serve as biomarkers of HIV-associated neurologic disorders. Since some markers will also be associated with cigarette smoking, tobacco use will be investigated for possible interfering effects on measurement of neurologic disease biomarkers.