PDF(Pubmed)
Abstract：
Diabetic nephropathy (DN) contributes to increased morbidity and mortality among patients with diabetes and presents a considerable global health challenge. However, reliable biomarkers of DN have not yet been established. Phosphorylated proteins are crucial for disease progression. However, their diagnostic potential remains unexplored. In this study, we used ultra-high-sensitivity quantitative phosphoproteomics to identify phosphoproteins in urinary extracellular vesicles (uEVs) as potential biomarkers of DN. We detected 233 phosphopeptides within the uEVs, with 47 phosphoproteins exhibiting significant alterations in patients with DN compared to those in patients with diabetes. From these phosphoproteins, we selected phosphorylated aquaporin-2 (p-AQP2[S256]) and phosphorylated glycogen synthase kinase-3β (p-GSK3β[Y216]) for validation, as they were significantly overrepresented in pathway analyses and previously implicated in DN pathogenesis. Both phosphoproteins were successfully confirmed through Phos-tag western blotting in uEVs and immunohistochemistry staining in kidney sections, suggesting that phosphoprotein alterations in uEVs reflect corresponding changes within the kidney and their potential as candidate biomarkers for DN. Our research proposes the utilization of phosphoproteins in uEVs as a liquid biopsy, presenting a highly feasible diagnostic tool for kidney disease.
摘要：
糖尿病肾病（DN）导致糖尿病患者的发病率和死亡率增加，并提出了相当大的全球健康挑战。然而,可靠的DN生物标志物尚未建立。磷酸化蛋白对疾病进展至关重要。然而,他们的诊断潜力仍未被开发。在这项研究中,我们使用超高灵敏度定量磷酸蛋白质组学鉴定尿细胞外囊泡(uEVs)中的磷蛋白作为DN的潜在生物标志物.我们在uEV中检测到233个磷酸肽，与糖尿病患者相比，DN患者中有47种磷蛋白表现出显着变化。从这些磷蛋白中，我们选择磷酸化水通道蛋白-2（p-AQP2[S256]）和磷酸化糖原合成酶激酶-3β（p-GSK3β[Y216]）进行验证，因为它们在通路分析中明显超标，并且以前与DN发病机制有关。通过uEV中的Phos-tagwestern印迹和肾脏切片中的免疫组织化学染色成功证实了这两种磷蛋白，提示uEV中的磷蛋白改变反映了肾脏内的相应变化及其作为DN候选生物标志物的潜力。我们的研究提出了在uEV中使用磷蛋白作为液体活检，为肾脏疾病提供了一种高度可行的诊断工具。
