Abstract:
Cataracts and glaucoma account for a high percentage of vision loss and blindness worldwide. Small extracellular vesicles (sEVs) are released into different body fluids, including the eye\'s aqueous humor. Information about their proteome content and characterization in ocular pathologies is not yet well established. In this study, aqueous humor sEVs from healthy individuals, cataracts, and glaucoma patients were studied, and their specific protein profiles were characterized. Moreover, the potential of identified proteins as diagnostic glaucoma biomarkers was evaluated. The protein content of sEVs from patients\' aqueous humor with cataracts and glaucoma compared to healthy individuals was analyzed by quantitative proteomics. Validation was performed by western blot (WB) and ELISA. A total of 828 peptides and 192 proteins were identified and quantified. After data analysis with the R program, 8 significantly dysregulated proteins from aqueous humor sEVs in cataracts and 16 in glaucoma showed an expression ratio ≥ 1.5. By WB and ELISA using directly aqueous humor samples, the dysregulation of 9 proteins was mostly confirmed. Importantly, GAS6 and SPP1 showed high diagnostic ability of glaucoma, which in combination allowed for discriminating glaucoma patients from control individuals with an area under the curve of 76.1% and a sensitivity of 65.6% and a specificity of 87.7%.
摘要:
白内障和青光眼占全球视力丧失和失明的比例很高。小细胞外囊泡(sEV)被释放到不同的体液中,包括眼睛的房水。关于其在眼部病理中的蛋白质组含量和表征的信息尚未得到很好的确定。在这项研究中,来自健康个体的房水sEV,白内障,对青光眼患者进行了研究,并对其特定的蛋白质谱进行了表征。此外,对鉴定的蛋白质作为诊断性青光眼生物标志物的潜力进行了评价.通过定量蛋白质组学分析了患有白内障和青光眼的患者房水与健康个体相比的sEV的蛋白质含量。通过蛋白质印迹(WB)和ELISA进行验证。鉴定并定量了总共828种肽和192种蛋白质。用R程序进行数据分析后,白内障中房水sEV中的8种明显失调的蛋白质和青光眼中的16种表达率≥1.5。通过WB和ELISA直接使用房水样品,9种蛋白质的失调大部分被证实。重要的是,GAS6和SPP1对青光眼有较高的诊断能力,结合起来可以将青光眼患者与对照个体区分开来,曲线下面积为76.1%,灵敏度为65.6%,特异性为87.7%。