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

 描述(申请人提供)：尽管大量的临床、实验室和影像证据表明，在接受联合抗逆转录病毒治疗(CART)的一些全身病毒抑制的艾滋病毒感染者中，中枢神经系统(CNS)持续异常，但这种扰动的潜在机制尚不清楚，包括病因、所涉及的细胞和组织，以及与潜在的艾滋病毒持久性的关系。在一个全球获得CART的时代，确定持续CART上中枢神经系统的扰动并确定潜在的干预措施是目前全球3500万艾滋病毒携带者最突出的神经学问题。目前的试点项目利用对外体microRNAs(exo-miRNA)在疾病发病机制中作用的认识，以进一步了解在长期抑制CART中检测到的HIV相关中枢神经系统异常的潜在过程。Exosome是通过供体细胞胞吐释放的膜纳米囊泡，其中包含调节靶细胞基因表达的小的非编码miRNAs(exo-miRNAs)。外源miRNAs在疾病的发病机制中发挥着重要作用，包括调节靶组织的免疫反应。此外，病毒的exo-miRNAs可能揭示了HIV的持久性，而其他特定的exo-miRNAs则表明了细胞起源。为了检验CNS中的exo-miRNA与CART治疗的HIV之间的关联，在目标1中，我们将通过对持续CART和未感染HIV的比较研究参与者的个人进行深入测序，横断面分析脑脊液(CSF)和血液中的exo-miRNA。我们假设，与炎症相关的脑脊液exo-miRNA在CART抑制的参与者中将更加丰富，并且与血液相比，脑脊液exo-miRNAs将被富含用于炎症中介miRNAs。我们将探索是否可以在脑脊液中检测到HIV衍生的exo-miRNAs，这表明HIV的持久性，以及早期和晚期启动CART对exo-miRNA图谱的影响。在目标2中，我们将研究外源miRNA图谱、中枢和血液中的免疫激活和耗竭以及神经认知能力之间的机制关系。我们假设，在脑脊液和血液中，与炎症相关的外源miRNAs将与单核细胞和T淋巴细胞的激活相关，这与流式细胞仪和免疫激活的可溶性标志物有关，而较差的神经心理测试表现将与脑脊液中miRNA的炎症特征相关。我们的长期目标是更好地了解HIV治疗后中枢神经系统紊乱的原因，以便开发有针对性的辅助疗法来改善HIV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.