Mechanisms of HIV persistence in the kidney

HIV 在肾脏中持续存在的机制

• 批准号: 10532574
• 负责人: Maria Blasi
• 金额: $ 68.11万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10532574
• 关键词: AIDS-Associated Nephropathy; APOL1 gene; Affect; Aftercare; Anti-Retroviral Agents; Archives; Automobile Driving; Autopsy; Base Sequence; Biopsy Specimen; Blood; CD4 Positive T Lymphocytes; Cell Death; Cell Proliferation; Cells; Chronic Disease; Clonal Expansion; Coculture Techniques; Consent; Data; Detection; Development; Epithelial Cells; Frequencies; Gene Expression; Gene Expression Profile; Genes; Genetic; Genetic Recombination; Genitourinary system; Genomic DNA; Growth; HIV; HIV Infections; HIV Seropositivity; HIV-1; Hypertrophy; Immune; In Vitro; Individual; Infection; Interruption; Kidney; Kidney Diseases; Kidney Transplantation; Location; Morbidity - disease rate; Pathogenesis; Pathology; Pathway interactions; Patients; Peripheral Blood Mononuclear Cell; Persons; Pharmaceutical Preparations; Phenotype; Phylogenetic Analysis; Plasma; Property; Protocols documentation; Renal tubule structure; Reporting; Research; Role; Sampling; Sequence Analysis; Site; Source; Specimen; System; T-Lymphocyte; Terminal Disease; Terminally Ill; Time; Tissues; Transplantation; Urine; Viral; Viral Load result; Viral Pathogenesis; Viral reservoir; Virus; Virus Latency; Virus Shedding; Work; antiretroviral therapy; chronic infection; experience; fight against; follow-up; in vivo; induced pluripotent stem cell; integration site; kidney allograft; kidney biopsy; kidney cell; kidney infection; macrophage; mortality; organ injury; podocyte; prospective; renal epithelium; risk variant; single-cell RNA sequencing; urogenital tract; viral RNA; viral detection; viral rebound

ABSTRACT Despite the dramatic improvement in HIV-associated morbidity and mortality with combination antiretroviral therapy (ART), HIV remains a chronic disease. The major barrier to HIV cure is the long-term persistence of multiple, latent viral reservoirs capable of reactivation in the absence of ART. Any effort to eradicate these reservoirs as part of a cure initiative requires understanding of the dynamics and control of HIV reactivation and replication in tissues and cells harboring the virus long-term. Our work has focused on understanding the mechanisms and implications of HIV infection of the kidney. We demonstrated that HIV infects renal tubule epithelial cells (RTEs) in vitro via direct contact with HIV-infected T cells and macrophages. Viral nucleic acid sequence analysis from in vivo derived RTEs compared to blood derived sequences demonstrated that the kidney represents a unique viral compartment. Furthermore, we showed that people with HIV (PWH) shed viral RNA in urine, and we have optimized approaches to detect and amplify HIV sequences from fresh and archived urine specimens. We found that some urine-derived HIV sequences were closely related to HIV sequences amplified from RTEs, supporting those cells as one of the sources of urine viruses. Viral detection in the urine allows for repeated sampling of the kidney compartment, which can be particularly useful in viral rebound studies. Additionally, in all of the PLWH we have analyzed so far, we amplified several identical HIV-1 sequences in urine, raising the possibility of clonal expansion of infected renal cells. Indeed, we recently reported that proliferation is one of the cellular fates observed in both actively and latently infected RTEs in vitro, together with hypertrophy and cell-death. Whether proliferation of infected renal epithelial cells contributes to HIV persistence in the kidney is unknown. The studies proposed here will define: 1) the long-term persistence of HIV in the kidney through the analysis of samples collected prospectively from PWH undergoing HIV+ to HIV+ kidney transplantation; 2) the reactivation potential of HIV in urine following ART interruption in terminally ill PWH who have consented to prospective follow-up as part of a rapid autopsy protocol; 3) the ability of patient-derived renal epithelial cells to carry replication competent virus; 4) the role of APOL1 kidney disease risk variants in RTE and podocyte infection; and 5) how HIV infection influences individual cell fate and potential for clonal expansion of infected RTEs. We hypothesize that renal epithelial cells serve as a long-term reservoir for HIV. Understanding the mechanisms of HIV persistence and reactivation in the kidney will inform cure strategies and further define renal pathogenesis in PLWH.

抽象的 尽管与抗逆转录病毒组合相关的艾滋病毒相关发病率和死亡率有了显着改善 治疗（ART），HI​​V仍然是一种慢性疾病。艾滋病毒治愈的主要障碍是长期的持久性 在没有艺术的情况下，能够重新激活的多种潜在病毒储存。消除这些的任何努力 作为治疗计划的一部分，水库需要了解HIV重新激活的动力学和控制 长期内携带病毒的组织和细胞中的复制。 我们的工作集中在理解肾脏感染的机制和含义上。我们 证明HIV通过直接接触HIV感染的T，在体外感染了肾小管上皮细胞（RTE） 细胞和巨噬细胞。与血液相比，体内衍生RTE的病毒核酸序列分析 衍生的序列表明肾脏代表一个独特的病毒室。此外，我们 表明患有艾滋病毒（PWH）的患者在尿液中脱离病毒RNA，我们采用了优化的方法来检测和 从新鲜和存档的尿液标本中扩增HIV序列。我们发现一些尿液衍生的艾滋病毒 序列与从RTE扩增的HIV序列密切相关，支持这些细胞作为其中之一 尿液病毒的来源。尿液中的病毒检测可以重复对肾脏室的采样， 这在病毒反弹研究中特别有用。此外，在所有PLWH中，我们都进行了分析 到目前为止，我们在尿液中扩增了几个相同的HIV-1序列，增加了克隆扩张的可能性 感染的肾细胞。确实，我们最近报道说增殖是两者中观察到的细胞命运之一 在体外积极和潜在地感染RTE，以及肥大和细胞死亡。是否扩散 感染的肾上皮细胞有助于肾脏中的HIV持续性。 这里提出的研究将定义：1）通过分析肾脏在肾脏中的长期持久性 从经历HIV+到HIV+肾脏移植的PWH收集的样品； 2）重新激活 在疾病的ART中断尿液中的尿液中的潜力 后续行动是快速尸检协议的一部分； 3）患者衍生的肾上皮细胞携带的能力 复制能力病毒； 4）APOL1肾脏疾病风险变异在RTE和足细胞感染中的作用； 5）HIV感染如何影响单个细胞命运和受感染RTE的克隆扩张的潜力。我们 假设肾上皮细胞是HIV的长期储备。了解机制 肾脏中的艾滋病毒持久性和重新激活将为治疗策略提供信息，并进一步定义肾脏发病机理 在PLWH中。