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长预计将呈指数级增长 糖尿病肾病(DKD)的增长。因此，阻止衰老相关肾脏变化的治疗干预措施 而DKD必须得到严格追查。适应不良的炎症导致DKD的发病，并促进炎症 细胞因子和趋化因子激活巨噬细胞，导致肾脏浸润，肾脏预后不良。然而， 炎症仍然是DKD中一种尚未解决的主要损伤途径。我们和其他人证明了 DKD患者和动物模型中的不适应性炎症与细胞衰老增加有关 和肾功能障碍。因此，通过衰老疗法，如小分子药物，可以延缓衰老 或细胞衍生成分，可能会阻止DKD和与年龄相关的肾脏恶化。因此，当务之急是 需要开发一种针对多方面发病机制(炎症和细胞)的治疗性武器 延缓衰老)，延长健康寿命。我们和其他人证明了间充质基质细胞 (MSCs)通过分泌含有以下成分的细胞外小泡(EV)来抑制炎症反应 生物活性物质，主要是microRNAs(MiRNAs)，并减少衰老负担和延长寿命 老化的老鼠模型。尽管MSC-EVS的治疗活性先前已被评估在衰老和 炎症性疾病模型，这些生物疗法对衰老中炎症和衰老的影响 DKD仍未被开发。我们的初步结果表明，MSC-EVS：(I)减少衰老途径， 巨噬细胞浸润与小鼠DKD肾脏损伤及(II)可负载小分子药物治疗 联合疗法。此外，我们最近的研究首次表明，抗衰老药物达沙替尼 和Qercetin，减少人体衰老细胞的丰度，改善衰老细胞引起的肾脏损伤 在小鼠模型中清除。我们推测，EVS具有抗炎和治疗作用。 老化的DKD，可以通过联合服用抗衰老药物来进一步增强。为了解决这一中心假设， 我们将确定肠道病毒miRNAs在体外对衰老和炎症的影响(目标1)，评估其影响 在衰老的DKD小鼠模型(目标2)中，从不同来源的EVS进行比较，并评估EVS的性能 Dasatinib和Qercetin的药物运载工具(目标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、细胞外小泡、感觉剂和药物方面具有互补的专业知识 送货。新方法包括独特的EV来源、EV分离方法和免疫图谱 技术将增强对EV介导的DKD治疗机制的了解，并可能推动临床 将EVS翻译为新的感官疗法、感觉剂的递送载体和/或联合疗法 减少糖尿病和年龄相关肾功能障碍的炎症和衰老途径。这些小说 治疗药物有可能改变疾病轨迹，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.