Mechanisms of HIV persistence in the kidney

HIV 在肾脏中持续存在的机制

基本信息

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
项目类别：
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

项目摘要

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.

抽象的尽管联合抗逆转录病毒药物显着改善了与艾滋病毒相关的发病率和死亡率治疗（ART），艾滋病毒仍然是一种慢性疾病。艾滋病毒治愈的主要障碍是长期持续存在多个潜伏病毒库能够在没有抗逆转录病毒治疗的情况下重新激活。任何消除这些现象的努力作为治疗计划的一部分，储存库需要了解艾滋病病毒再激活的动态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艾滋病毒肾脏感染的机制和影响。我们证明 HIV 通过与感染 HIV 的 T 细胞直接接触而在体外感染肾小管上皮细胞 (RTE) 细胞和巨噬细胞。与血液相比，体内 RTE 的病毒核酸序列分析衍生序列表明肾脏代表了独特的病毒区室。此外，我们研究表明，艾滋病毒感染者 (PWH) 会在尿液中排出病毒 RNA，我们已经优化了检测和检测方法从新鲜和存档的尿液样本中扩增 HIV 序列。我们发现一些尿液来源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。体外主动和潜伏感染 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艾滋病毒在肾脏中持续存在的原因尚不清楚。这里提出的研究将定义：1）通过分析 HIV 在肾脏中的长期持续存在从接受 HIV+ 到 HIV+ 肾移植的 PWH 前瞻性收集样本； 2）重新激活同意前瞻性治疗的绝症感染者在 ART 中断后尿液中 HIV 的可能性作为快速尸检方案的一部分进行后续行动； 3）患者来源的肾上皮细胞的携带能力有复制能力的病毒； 4) APOL1肾病风险变异在RTE和足细胞感染中的作用； 5) HIV 感染如何影响个体细胞命运以及受感染 RTE 克隆扩增的潜力。我们假设肾上皮细胞是 HIV 的长期储存库。了解其机制 HIV 在肾脏中的持续存在和重新激活将为治疗策略提供信息并进一步明确肾脏发病机制在艾滋病毒感染者中。