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.

摘要 尽管联合抗逆转录病毒治疗在艾滋病毒相关的发病率和死亡率方面有显著改善， 尽管艾滋病毒治疗（ART）的效果不佳，但它仍然是一种慢性疾病。艾滋病毒治愈的主要障碍是长期存在的 在没有ART的情况下能够重新激活的多个潜伏的病毒储库。 作为治疗计划的一部分，需要了解HIV再激活的动力学和控制 并在长期携带病毒的组织和细胞中复制。 我们的工作重点是了解HIV感染肾脏的机制和影响。我们 证明HIV通过与HIV感染的T细胞直接接触感染体外肾小管上皮细胞（RTEs）， 细胞和巨噬细胞。与血液相比，来自体内来源的RTE的病毒核酸序列分析 衍生的序列表明肾脏代表独特的病毒区室。而且我们 研究表明，艾滋病毒感染者（PWH）在尿液中释放病毒RNA，我们已经优化了检测和 从新鲜和存档的尿液标本中扩增HIV序列。我们发现一些尿液中的艾滋病病毒 序列与从RTEs扩增的HIV序列密切相关，支持这些细胞是 尿液病毒的来源。尿液中的病毒检测允许对肾室进行重复采样， 这在病毒反弹研究中特别有用。此外，在我们分析的所有PLWH中， 到目前为止，我们在尿液中扩增了几个相同的HIV-1序列，提高了HIV-1克隆扩增的可能性。 感染的肾细胞事实上，我们最近报道说，增殖是在两种细胞中观察到的细胞命运之一。 体外活跃和潜伏感染RTE，并伴有肥大和细胞死亡。是否扩散 受感染的肾上皮细胞导致HIV在肾脏中的持续存在是未知的。 本文提出的研究将定义：1）通过分析肾脏中的HIV长期持续存在， 从接受HIV+到HIV+肾移植的PWH前瞻性收集的样本; 2）重新激活 已同意接受前瞻性抗逆转录病毒治疗的晚期威尔斯亲王医院患者中断抗逆转录病毒治疗后尿液中HIV的可能性 随访作为快速尸检方案的一部分; 3）患者来源的肾上皮细胞携带 4）APOL 1肾病风险变体在RTE和足细胞感染中的作用; 以及5）HIV感染如何影响个体细胞命运和受感染RTE的克隆扩增潜力。我们 假设肾上皮细胞是HIV长期储存库。了解的机制 HIV在肾脏中的持续存在和再活化将为治疗策略提供信息，并进一步确定肾脏发病机制 在PLWH。