Viral dynamics of rebound and reservoir HIV species in IPSC-derived myeloid cells

IPSC 衍生骨髓细胞中反弹和储存 HIV 物种的病毒动态

• 批准号: 10684814
• 负责人: Cagla Akay Espinoza
• 金额: $ 20.31万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10684814
• 关键词: Address; Adherence; Adult; Anatomy; Binding; Blood; Blood Vessels; Cell Lineage; Cells; Central Nervous System; Cerebrospinal Fluid; Characteristics; Conceptions; Data; Development; Exhibits; Fibroblasts; HIV; Human; Infection; Interferons; Interruption; Knowledge; Macrophage; Microglia; Modeling; Molecular Cloning; Mus; Myelogenous; Myeloid Cells; Patients; Phenotype; Productivity; Resistance; Role; Signal Transduction; Site; Source; Surface; System; T-Lymphocyte; Technology; Testing; Tissues; Toxic effect; Tropism; Up-Regulation; Variant; Viral; Viral reservoir; Viremia; Virus; Virus Replication; antiretroviral therapy; chronic infection; density; follow-up; in vitro Model; in vivo; induced pluripotent stem cell; lymph nodes; monocyte; novel strategies; pressure; trafficking; transmission process; variant detection; viral DNA; viral rebound; virus identification; Address; Adherence; Adult; Anatomy; Binding; Blood; Blood Vessels; Cell Lineage; Cells; Central Nervous System; Cerebrospinal Fluid; Characteristics; Conceptions; Data; Development; Exhibits; Fibroblasts; HIV; Human; Infection; Interferons; Interruption; Knowledge; Macrophage; Microglia; Modeling; Molecular Cloning; Mus; Myelogenous; Myeloid Cells; Patients; Phenotype; Productivity; Resistance; Role; Signal Transduction; Site; Source; Surface; System; T-Lymphocyte; Technology; Testing; Tissues; Toxic effect; Tropism; Up-Regulation; Variant; Viral; Viral reservoir; Viremia; Virus; Virus Replication; antiretroviral therapy; chronic infection; density; follow-up; in vitro Model; in vivo; induced pluripotent stem cell; lymph nodes; monocyte; novel strategies; pressure; trafficking; transmission process; variant detection; viral DNA; viral rebound; virus identification

Eradicative HIV cure requires elimination of virus from all tissues, including the central nervous system (CNS), and a thorough understanding of HIV reservoirs that harbor intact and/or replication-competent virus and give rise or support rebound viremia upon treatment interruption is paramount. As long-lived CNS myeloid cells, namely microglia and resident macrophages are an HIV reservoir. While HIV enters the CNS during primary viremia and the predominant HIV variants isolated soon after infection exhibit T-cell tropism, HIV variants that infect cells with lower surface CD4 density, such as microglia, are found later during chronic infection, supported by HIV viral DNA isolated from perivascular and CNS myeloid cells and sustained HIV replication in macrophages independently of T cells in humanized myeloid-only mice. HIV from a T-cell source has also been detected in the cerebrospinal fluid (CSF) of PLWH on suppressive ART. HIV variants detected in the CSF are likely a result of a combination of independent replication of HIV compartmentalized in the CNS in CD4+ T and/or myeloid­ lineage cells and HIV trafficking via T cells or monocytes or as free virus from the periphery. Technical and anatomical constraints limit the comprehensive examination of (i) cellular and viral factors involved in HIV replication in the CNS and (ii) selection pressures that give rise to rebound viruses in the correct cellular context, especially in microglia. Among the reasons are limited availability of freshly isolated primary human microglia, which nonetheless rapidly lose key in vivo features, and the paucity of in vitro models that successfully recapitulate key microglial characteristics. We will use human induced pluripotent stem cell-derived microglia (iMg) differentiated from adult human fibroblasts as a tractable system to examine host and viral determinants governing rebound virus replication in CNS myeloid cells with detailed analyses that are not possible with currently available models. Two recent developments regarding rebound viruses has led to the conception of this proposal. First, in a recent study examining key viral phenotypes in patients undergoing analytical treatment interruption (ATI), our collaborator Dr. Bar found that viruses that rebounded after ATI exhibited higher IFN-1 resistance than any other viruses, including transmitted founder (TF) viruses and reservoir viruses, among viruses isolated from the blood throughout the longitudinal follow-up of patients. The origins of these uniquely IFN-1-resistant rebound viruses remain unclear. Persistent upregulation of IFN-1 signaling in the CNS of PLWH despite suppressive ART raises the possibility that selection pressures might lead to the establishment of a productive reservoir of IFN-1-resistant virus within CNS myeloid-lineage cells. Second, our preliminary data suggest that infectious molecular clones (IMCs) of some rebound viruses identified by Dr. Bar may replicate better than the IMCs of TF viruses in iMg. In this proposal, we will use iMg with a panel of well-characterized IMCs of TF, reservoir, and rebound viruses to test our hypothesis that CNS myeloid cells may be a site for replication of IFN-1-resistant rebound viruses.

根除的HIV治疗需要从所有组织中消除病毒，包括中枢神经系统（CNS），以及对具有完整和/或复制能力的病毒的艾滋病毒储层的透彻理解，并在干扰时产生或支持反弹病毒血症。长期寿命的CNS髓样细胞（即小胶质细胞）和居民巨噬细胞是HIV储层。 While HIV enters the CNS during primary viremia and the predominant HIV variants isolated soon after infection exhibit T-cell tr​​opism, HIV variants that infected cells with lower surface CD4 density, such as microglia, are found later during chronic infection, supported by HIV viral DNA isolated from perivascular and CNS myeloid cells and sustained HIV replication in macrophages independently of T cells in humanized仅髓样小鼠。在抑制作用的PLWH的脑脊液（CSF）中也发现了来自T细胞源的HIV。 CSF中检测到的艾滋病毒变体可能是CD4+ T和/或髓样谱系细胞中CNS中的HIV独立复制的结果，以及通过T细胞或单核细胞或单核细胞或从外周的游离病毒进行的HIV运输。技术和解剖学约束限制了CNS中涉及HIV复制的（I）在正确的细胞环境中引起反弹病毒的选择压力的（ii）选择压力的（ii）选择压力的全面检查。原因之一是新鲜分离的原发性小胶质细胞的可用性有限，尽管如此，这些小胶质细胞却迅速失去了体内特征的关键特征，以及成功概括关键小胶质细胞特征的体外模型的匮乏。我们将使用人类诱导的多能干细胞衍生的小胶质细胞（IMG）与成年人成纤维细胞区分开来作为一种可拖动系统来检查宿主，病毒决定了CNS髓样细胞中的反弹病毒复制，并具有目前可用的模型无法使用的详细分析。关于反弹病毒的最新发展导致了该提案的概念。首先，在最近的一项研究中检查了接受分析治疗中断（ATI）的患者中的关键病毒表型（ATI）的研究中，我们的合作者Bar博士发现，ATI后反弹的病毒暴露于与其他任何病毒相比，包括从纵向培养患者中分离出的病毒中的任何其他病毒，包括传播的创建者（TF）病毒和储层病毒，与病毒分离出来。这些独特的IFN-1反弹病毒的起源尚不清楚。在PLWH目的地抑制ART的中枢神经系统中，IFN-1信号传导的持续上调提高了选择压力可能导致在CNS髓样细胞内建立IFN-1耐药性病毒的生产性储层的可能性。其次，我们的初步数据表明，BAR博士确定的一些反弹病毒的传染性分子克隆（IMC）可能比IMG中TF病毒的IMC更好地复制。在此提案中，我们将使用IMG和一组TF，储层和反弹病毒的特征良好的IMC来测试我们的假设，即CNS髓样细胞可能是复制IFN-1耐药反弹病毒病毒的位点。