Elucidating the Molecular Mechanisms that Mediate DKD Progression in Patients Living with HIV

阐明介导 HIV 感染者 DKD 进展的分子机制

• 批准号: 10364063
• 负责人: John Cijiang He
• 金额: $ 85.86万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-01 至 2026-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10364063
• 关键词: Aging; Animal Model; Archives; Automobile Driving; Biopsy; Cells; Chronic; Chronic Kidney Failure; Clinical Data; Collaborations; Complement; Data; Data Set; Development; Diabetes Mellitus; Diabetic Nephropathy; Diabetic mouse; Disease Progression; Dose; Doxycycline; End stage renal failure; Epithelial Cells; Exposure to; Formalin; Gene Expression; Gene Expression Profile; Genes; Goals; HIV; HIV Infections; HIV Seropositivity; HIV therapy; HIV-1; Hepatitis C virus; Human; Hypertension; Immune; In Situ Hybridization; Inflammation; Injury; Injury to Kidney; Kidney; Kidney Diseases; Kidney Transplantation; Knowledge; Leukocytes; Longitudinal Studies; Mediating; Modeling; Molecular; Mus; Online Systems; Organ; PTPRC gene; Paraffin Embedding; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacology; Plasma; Population; Prevalence; Process; Publishing; RNA; Regimen; Reporting; Sample Size; Scientist; Slide; Streptozocin; System; T-Lymphocyte; Tissues; Transgenic Mice; Transgenic Organisms; Tubular formation; United States; Veterans; Viral Load result; Viremia; age related; antiretroviral therapy; base; cohort; comorbidity; diabetic; diabetic patient; efficacy validation; genetic signature; kidney biopsy; kidney cell; mouse model; new therapeutic target; non-diabetic; novel therapeutics; personalized medicine; podocyte; response; single-cell RNA sequencing; small molecule; targeted treatment; transcriptomics; web portal

SUMMARY With the aging of the HIV-infected population, and their prolonged exposure to cART regimens that may promote the development of diabetes, the prevalence of diabetic kidney disease (DKD) is increasing in patients living with HIV (PLWH). In two recent studies, we and others have confirmed the additive/synergistic effects of HIV infection and diabetes on the progression of DKD. Studies have demonstrated that HIV can infect kidney cells, and that the kidney serves as a reservoir for HIV replication. Recent findings suggest that HIV is present in the kidney even in cART-treated HIV patients. To determine how chronic HIV infection in patients with low or undetectable viral load aggravates DKD, we generated a doxycycline-inducible kidney cell-specific HIV transgenic mouse the expresses low levels of HIV genes. With this model, we confirmed that low level of HIV gene expression did not cause kidney injury alone. However, aggravated DKD was observed after induction of diabetes, suggesting an additive/synergistic effect of HIV and diabetes on DKD progression (Feng J. Kidney Int. 2021). For this project, we will further investigate the molecular mechanisms by which chronic HIV infection induces the progression of DKD in both mouse models and human patients with HIV to accelerate the discovery of new drug targets and therapeutics to treat these patients. Toward this goal, we propose the following specific aims: Aim 1: Perform comprehensive bulk and single-cell RNA-sequencing of the kidneys and kidney immune cells from the mice with both diabetes and low HIV gene expression as compared to control mice, and mice with HIV or diabetes alone. Aim 2: Perform formalin fixed paraffin embedded slides-based RNA-capture sequencing, scRNA-seq, immunostaining, and in-situ hybridization of kidneys from patients with DKD alone, and from those with both HIV infection and DKD. Aim 3: Integrate these datasets together with publicly available datasets to identify gene expression signatures mediating the additive/synergistic effects of HIV and diabetes on DKD progression. Utilize the Connectivity Mapping approach to identify potential drugs and small molecules which could reverse these gene signatures similarly to the approach described in our recent study (Zhang L. Diabetes 2020). Develop a web-portal to serve and interact with all project data. Aim 4: Validate predicted drugs from aim 3 in the animal model described in the aim 1. Overall, this project will enable us to identify potential new mechanisms, new drug targets, and potential new therapeutics to halt the progression of DKD in PLWH.

概括 随着 HIV 感染人群的老龄化，以及他们长期接触 cART 治疗方案，可能会促进 随着糖尿病的发展，患有糖尿病的患者中糖尿病肾病（DKD）的患病率正在增加 艾滋病毒（艾滋病病毒感染者）。在最近的两项研究中，我们和其他人证实了艾滋病毒感染的累加/协同效应 和糖尿病对 DKD 进展的影响。研究表明，艾滋病毒可以感染肾细胞，并且 肾脏是艾滋病毒复制的储存库。最近的研究结果表明艾滋病毒存在于肾脏中 即使在接受 cART 治疗的 HIV 患者中也是如此。确定慢性 HIV 感染率低或无法检测的患者的情况 病毒载量加重 DKD，我们制备了强力霉素诱导的肾细胞特异性 HIV 转基因小鼠 表达低水平的 HIV 基因。通过这个模型，我们证实低水平的 HIV 基因表达并没有 单独造成肾损伤。然而，在诱导糖尿病后观察到 DKD 加重，这表明 HIV 和糖尿病对 DKD 进展的累加/协同作用（Feng J. Kidney Int. 2021）。对于这个项目， 我们将进一步研究慢性HIV感染诱发病情进展的分子机制 DKD 在小鼠模型和人类 HIV 患者中进行，以加速新药靶点的发现和 治疗这些患者的疗法。为了实现这一目标，我们提出以下具体目标： 目标 1：执行 对小鼠的肾脏和肾脏免疫细胞进行全面的批量和单细胞 RNA 测序 与对照小鼠以及仅患有艾滋病毒或糖尿病的小鼠相比，糖尿病和低艾滋病毒基因表达。 目标 2：执行基于福尔马林固定石蜡包埋载玻片的 RNA 捕获测序、scRNA-seq、 对单纯 DKD 患者和同时感染 HIV 患者的肾脏进行免疫染色和原位杂交 感染和 DKD。目标 3：将这些数据集与公开可用的数据集整合在一起以识别基因 介导 HIV 和糖尿病对 DKD 进展的累加/协同效应的表达特征。利用 连接映射方法来识别可以逆转这些的潜在药物和小分子 基因特征与我们最近的研究中描述的方法类似（Zhang L. Diabetes 2020）。开发一个 用于服务所有项目数据并与之交互的门户网站。目标 4：在动物身上验证目标 3 预测的药物 目标 1 中描述的模型。总体而言，该项目将使我们能够识别潜在的新机制、新药物 目标，以及阻止 PLWH 中 DKD 进展的潜在新疗法。