CSF & Blood Exosomal microRNAs, Immune Responses, and HAND in ART Suppressed HIV

 描述（由申请方提供）：尽管有大量临床、实验室和影像学证据表明，在接受抗逆转录病毒联合治疗（cART）的一些全身性病毒抑制的HIV感染个体中，中枢神经系统（CNS）持续异常，但对这种扰动的潜在机制仍知之甚少，包括病因学、涉及的细胞和组织以及与HIV潜在持续性的关系。在全球越来越多地获得cART的时代，定义CNS对持续cART的干扰并确定潜在的干预措施是目前全球3500万艾滋病毒感染者最突出的神经学问题。目前的试点项目利用了对外来体微小RNA（exo-miRNA）在疾病发病机制中的作用的认识，以进一步了解长期抑制性cART检测到的HIV相关CNS异常的潜在过程。外泌体是通过供体细胞的胞吐作用释放的膜纳米囊泡，其含有调节靶细胞中基因表达的小的非编码miRNA（exo-miRNA）。exo-miRNAs在疾病的发病机制中起重要作用，包括调节靶组织的免疫应答。此外，病毒exo-miRNA可以揭示HIV持久性，并且其他特异性exo-miRNA指示细胞起源。为了检查CNS中的exo-miRNA与cART治疗的HIV之间的关联，在目的1中，我们将通过对持续cART和HIV未感染的比较研究参与者的个体进行深度测序，对脑脊液（CSF）和血液中的exo-miRNA进行横断面分析。我们假设与炎症相关的CSF exo-miRNA在cART抑制的参与者中更丰富，并且与血液相比，CSF exo-miRNA将富集炎症介导的miRNA。我们将探索是否可以在CSF中检测到HIV衍生的exo-miRNA，这表明HIV持续存在，以及早期与晚期启动cART对exo-miRNA谱的影响。在目标2中，我们将研究exo-miRNA谱、CNS和血液中的免疫激活和耗竭以及神经认知表现之间的机制关系。我们假设，在CSF和血液中，与炎症相关的exo-miRNAs将与通过流式细胞术和免疫活化的可溶性标记物测量的单核细胞和T淋巴细胞的活化相关，并且较差的神经心理学测试表现将与CSF中的miRNA的炎症特征相关。我们的长期目标是提供更好的理解CNS扰动的原因在治疗艾滋病毒，以开发有针对性的治疗，以改善艾滋病毒相关的神经认知障碍。

 DESCRIPTION (provided by applicant): Despite extensive clinical, laboratory, and imaging evidence that the central nervous system (CNS) is persistently abnormal in some HIV-infected individuals with systemic viral suppression on combination antiretroviral therapy (cART), the underlying mechanisms of this perturbation are poorly understood, including etiologies, cells and tissues involved, and relation to potential persistence of HIV. In an era of increasing global access to cART, defining perturbations of the CNS on sustained cART and identifying potential interventions is the most salient neurologic issue for the 35 million people currently living worldwide with HIV. The current pilot project exploits recognition of the role of exosomal micro-RNAs (exo-miRNA) in disease pathogenesis to further our understanding of the processes underlying HIV related CNS abnormality detected on long-term suppressive cART. Exosomes are membrane nanovesicles released via exocytosis of donor cells that contain small non-coding miRNAs (exo-miRNAs) that regulate gene expression in target cells. Exo-miRNAs play important roles in disease pathogenesis including regulating immune responses in target tissues. Furthermore, viral exo- miRNAs may reveal HIV persistence, and other specific exo-miRNAs indicate cellular origins. To examine the associations between exo-miRNA in the CNS and cART treated HIV, in Aim 1 we will cross-sectionally profile exo-miRNA in cerebrospinal fluid (CSF) and blood by deep sequencing in individuals on sustained cART and HIV-uninfected comparison study participants. We hypothesize that CSF exo-miRNA associated with inflammation will be more abundant in cART-suppressed participants and that CSF exo-miRNAs will be enriched for inflammatory-mediating miRNAs compared to blood. We will explore whether HIV-derived exo-miRNAs can be detected in CSF, suggesting HIV persistence, and the impact of early versus later initiation of cART on exo-miRNA profiles. In Aim 2 we will investigate the mechanistic relationships between exo-miRNA profiles, immune activation and exhaustion in the CNS and blood, and neurocognitive performance. We hypothesize that in CSF and blood, exo-miRNAs linked to inflammation will correlate with activation of monocytes and T lymphocytes as measured by flow cytometry and soluble markers of immune activation, and that poorer neuropsychological testing performance will associate with inflammatory profiles of miRNA in CSF. Our long-term goal is to provide improved understanding of the causes of CNS perturbation in treated HIV in order to develop targeted adjunctive therapies to ameliorate HIV associated neurocognitive disorder.