Targeting_the_Source_of_Persistent_HIV_Viremia

目前的抗逆转录病毒疗法不能治愈，也不能根除HIV-1病毒血症。相反，病毒会持续存在 只是在治疗过程中被抑制了。这种持续性病毒血症的来源和动态目前是 未知。在联合抗逆转录病毒治疗期间确定持续性病毒血症的来源将有助于 未来旨在根除病毒的努力。为了解决这些重要问题，目前的研究提出了两个 假设。首先，为了确定接受抑制治疗的患者持续病毒血症的来源，我们必须 比较患者血浆中的HIV种群与脑脊液(CSF)细胞中的HIV种群 从外周血液和从其他组织中分离出来的细胞。其次，激活潜伏感染艾滋病毒的细胞将 诱导整合的艾滋病毒基因组的表达，使其容易受到免疫介导的杀伤和 心理治疗。为了检验这些假设，我们试图实现三个具体目标。首先，我们将描述HIV-1的特征 血浆、脑脊液和来自外周血、肠道的广泛HIV感染细胞中的遗传群体 患者的相关淋巴组织(GALT)、淋巴组织(LNT)和骨髓组织(BMT) 采用单基因组测序和单前病毒测序等方法进行抑制治疗。这个 血浆来源的病毒RNA序列与细胞内病毒DNA序列的亲缘关系 将会被确定。从血浆到不同细胞和组织中HIV种群的比较 隔室将使我们能够调查和寻找持久的来源和遗传动力学 接受抑制治疗的患者中的病毒血症。对于我们的第二个目标，我们将采用同行评议并发表 基于连接的分子工具，称为邻近连接探针，用于专门标记和测量HIV病毒粒子 并确定这些粒子的细胞来源。这对于确定细胞谱系很重要。 在抑制治疗期间产生病毒粒子。第三，我们将使用新的DNA甲基化抑制剂，蛋白质 甲基化或组蛋白去乙酰化刺激细胞HIV转录，从而释放HIV 潜伏感染记忆的CD4+T细胞，使病毒容易受到抗逆转录病毒药物的攻击。要做到这一点 目的：我们将培养接受抑制治疗的患者的CD4+记忆T细胞，并将这些细胞暴露于 新的抑制剂。该研究计划将汇集来自欧洲和欧洲的不同的多国专家小组 美国专门从事艾滋病毒翻译研究、艾滋病毒临床治疗、分子遗传学和 抗病毒药物开发。

Current antiretroviral therapy is not curative and does not eradicate HIV-1 viremia. Rather, the virus persists and is merely suppressed during therapy. The source and dynamics of this persistent viremia is currently unknown. Determining the source of persistent viremia during combination antiretroviral therapy will inform future efforts aimed at viral eradication. To address these important issues the current study puts forward two hypotheses. First, to identify the source of persistent viremia in patients on suppressive therapy we must compare HIV populations in patient plasma to the HIV populations found in the cerebrospinal fluid (CSF), cells from peripheral blood and cells isolated from other tissues. Second, activating the latently HIV-infected cell will induce the expression of the integrated HIV genome making it vulnerable to immune-mediated killing and therapy. To test these hypotheses we seek to achieve three specific aims. First, we will characterize the HIV-1 genetic populations in plasma, CSF and a broad spectrum of HIV-infected cells from peripheral blood, gut associated lymphoid tissue (GALT), lymph node tissue (LNT), and bone marrow tissue (BMT) from patients on suppressive therapy using such methods as single-genome sequencing and single-proviral sequencing. The phylogenetic relatedness of the plasma-derived viral RNA sequences and intracellular viral DNA sequences will be determined. The comparison of HIV populations from plasma to different cellular and tissue compartments will allow us to investigate and seek to identify the source and genetic dynamics of persistent viremia in patients on suppressive therapy. For our second aim, we will apply peer-reviewed and published ligation-based molecular tools known as proximity ligation probes to specifically label and measure HIV virions and identify the cellular source of these particles. This will be important for identifying the cell lineage producing virions during suppressive therapy. Third, we will use new inhibitors of DNA methylation, protein methylation or histone deacetylation to stimulate cellular HIV transcription and therefore release of HIV from latently infected memory CD4+ T-cells, making the virus vulnerable to antiretroviral drugs. To accomplish this aim, we will culture CD4+ memory T-cells from patients on suppressive therapy and expose these cells to the new inhibitors. This research plan will bring together a diverse, multinational group of experts from Europe and United States specializing in HIV translational research, HIV clinical treatment, molecular genetics, and antiviral drug development.