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

总结 随着艾滋病毒感染人群的老龄化，以及他们长期接触cART方案， 随着糖尿病的发展，糖尿病肾病（DKD）的患病率在患有糖尿病的患者中正在增加。 艾滋病毒（艾滋病毒携带者）。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叠加/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， 肾脏是HIV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， 即使是接受cART治疗的HIV患者。确定慢性HIV感染患者如何低或检测不到 病毒载量增加DKD，我们建立了一个多西环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。利用这个模型，我们证实了HIV基因的低水平表达并不影响 会导致肾损伤。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/协同效应（Feng J. Kidney Int. 2021）。对于这个项目， 我们将进一步研究慢性HIV感染诱导慢性炎症进展的分子机制。 在小鼠模型和人类HIV患者中进行DKD，以加速发现新的药物靶点， 治疗这些患者的方法。为了实现这一目标，我们提出了以下具体目标：目标1： 对来自患有以下疾病的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： 与对照小鼠和仅患有HIV或糖尿病的小鼠相比，糖尿病和HIV基因表达低。 目的2：进行基于福尔马林固定石蜡包埋载玻片的RNA捕获测序，scRNA-seq， 免疫染色和原位杂交检测DKD单独患者和HIV双阳性患者的肾脏 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 表达特征介导HIV和糖尿病对DKD进展的累加/协同作用。利用 连接性映射方法，以确定潜在的药物和小分子，可以扭转这些 基因签名类似于我们最近的研究中描述的方法（Zhang L.糖尿病2020）。开发一个 门户网站，提供所有项目数据并与之交互。目的4：在动物中验证目的3中预测的药物 目标1中描述的模型。总的来说，这个项目将使我们能够识别潜在的新机制，新的药物， 靶点和潜在的新疗法来阻止PLWH中DKD的进展。

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.