Extracellular vesicle-based senotherapeutics for aging diabetic kidneydisease

项目摘要 受肥胖和糖尿病困扰的老龄化人口的增加预计将呈指数级增长， 糖尿病肾病（DKD）的发病率。因此，阻止与衰老相关的肾脏变化的治疗干预措施 必须严格执行DKD。适应不良炎症驱动DKD发作， 细胞因子和趋化因子激活巨噬细胞，导致肾浸润和肾预后不良。然而， 炎症仍然是DKD中主要的未解决的损伤途径。我们和其他人证明， DKD患者和动物模型中的适应不良炎症与细胞衰老增加相关 和肾功能障碍因此，通过衰老治疗剂（例如小分子药物）来减少衰老， 或细胞衍生成分，可以阻止DKD和年龄相关的肾脏恶化。因此迫切 需要开发一种针对多方面发病机制（炎症和细胞 DKD的衰老），以延长健康寿命。我们和其他人证明了间充质基质细胞 骨髓间充质干细胞（MSC）通过分泌含有细胞外囊泡（EV）的细胞外囊泡（EV）来抑制炎症反应。 生物活性货物，主要是microRNA（miRNAs），并减少衰老负担和延长寿命， 衰老小鼠模型。尽管先前已经在衰老和衰老中评估了MSC-EV的治疗活性， 炎症性疾病模型，这些生物治疗剂对炎症和衰老的影响， DKD仍然未被探索。我们的初步结果表明，MSC-EV：（i）减少衰老途径， 巨噬细胞浸润和小鼠DKD中的肾损伤，和（ii）可以负载小分子药物， 联合治疗此外，我们最近的研究首次证明，衰老清除药物达沙替尼 和槲皮素，减少人类衰老细胞的丰度，并改善衰老细胞 小鼠模型中的清除率。我们假设电动汽车具有抗炎和镇静作用， 衰老DKD，其可通过共递送衰老清除药物而进一步增强。为了解决这个中心假设， 我们将在体外确定EV miRNAs对衰老和炎症的作用（目的1），评估其作用， 在衰老DKD小鼠模型（目的2）中检测来自几种来源的EV，并评估EV的性能， 达沙替尼和槲皮素的药物递送载体（目的3）。为了确保这项研究的技术成功，我们 我组建了一个团队，在衰老，DKD，细胞外囊泡，senolytics和药物方面具有互补的专业知识 交付.包括独特EV来源、EV分离方法和免疫分析的新方法 技术将提高对EV介导的DKD治疗机制的认识， 将EV转化为新的衰老治疗剂、用于衰老清除剂的递送载体和/或组合疗法， 减少糖尿病和年龄相关性肾功能障碍的炎症和衰老途径。这些新颖 治疗方法有可能改变疾病轨迹并延长老年DKD患者的健康寿命。

Project Summary The rise in an aging population plagued by obesity and diabetes mellitus is projected to render exponential growth in diabetic kidney disease (DKD). Hence, therapeutic interventions that halt aging-related kidney changes and DKD must be rigorously pursued. Maladaptive inflammation drives DKD onset, and proinflammatory cytokines and chemokines activate macrophages leading to kidney infiltration and poor renal prognosis. Yet, inflammation remains a major unaddressed injurious pathway in DKD. We and others demonstrated that maladaptive inflammation in DKD patients and animal models is associated with increased cellular senescence and kidney dysfunction. Thus, decreasing senescence through senotherapeutics, such as, small molecule drugs or cell-derived components, may halt DKD and age-related kidney deterioration. Therefore, there is an urgent need to develop a therapeutic armamentarium targeting the multifaceted pathogenesis (inflammation and cell senescence) of DKD to extend healthy lifespan. We and others demonstrated that mesenchymal stromal cells (MSCs) suppress inflammatory responses through secretion of extracellular vesicles (EVs) containing biologically active cargo, primarily microRNAs (miRNAs), and reduce senescent burden and extend lifespan in aging mouse models. Although the therapeutic activity of MSC-EVs has previously been assessed in aging and inflammatory disease models, the effects of these biotherapeutics on inflammation and senescence in aging DKD remain unexplored. Our preliminary results indicate that MSC-EVs: (i) reduce senescence pathways, macrophage infiltration, and kidney injury in murine DKD and (ii) can be loaded with small molecule drugs for combination therapy. Additionally, our recent study demonstrated, for the first time, that senolytic drugs, dasatinib and quercetin, reduce senescent cell abundance in humans and improve kidney injury following senescent cell clearance in mouse models. We hypothesize that EVs have anti-inflammatory and senotherapeutic effects in aging DKD, that can be further enhanced by co-delivery of senolytic drugs. To address this central hypothesis, we will determine the effects of EV miRNAs on senescence and inflammation in vitro (Aim 1), assess the effects of EVs from several sources in a mouse model of aging DKD (Aim 2), and evaluate the performance of EVs as drug delivery vehicles for dasatinib and quercetin (Aim 3). To ensure the technical success of this study, we have assembled a team with complementary expertise in aging, DKD, extracellular vesicles, senolytics, and drug delivery. Novel approaches encompassing unique EV sources, EV isolation methods, and immunoprofiling technology will enhance knowledge of EV-mediated therapeutic mechanisms in DKD and may advance clinical translation of EVs as novel senotherapeutics, delivery vehicles for senolytics, and/or combination therapies to reduce inflammatory and senescence pathways in diabetic and age-related kidney dysfunction. These novel therapeutics hold potential to alter disease trajectory and extend the healthy lifespan in those with aging DKD.