Department of Ophthalmology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel.
Invest Ophthalmol Vis Sci. 2021 Aug 2;62(10):18. doi: 10.1167/iovs.62.10.18.
Age-related macular degeneration (AMD) is associated with altered gene and protein expression in the retina. We characterize the aqueous humor (AH) proteome in AMD to gain insight into the pathogenesis of the disease and identify potential biomarkers.
AH was collected from age and gender matched neovascular AMD (nvAMD; n = 10) patients and controls (n = 10). AH was pooled to create two samples (nvAMD and control), followed by intensity-based label-free quantification (MS1). Functional and bioinformatic analysis were then performed. A validation set (20 controls, 15 atrophic AMD and 15 nvAMD) was tested via multiplex ELISA for nine differentially expressed proteins according to the MS1 findings.
MS1 identified 674 proteins in the AH. 239 proteins were upregulated in nvAMD (nvAMD/control > 2, peptide tags (PT) > 2), and 86 proteins were downregulated (nvAMD/control < 0.5, PT > 2). Functional analysis of proteins upregulated in AMD demonstrated enrichment for platelet degranulation (enrichment score (ES):28.1), negative regulation of endopeptidase activity (ES:18.8), cellular protein metabolic process (ES:11.8), epidermal growth factor-like domain (ES:10.3), sushi/SCR/CCP (ES:10.1), and complement/coagulation cascades (ES:9.2). AMD protein clusters were upregulated for 3/6 (χ2 < 0.05 compared to randomization). Validation via ELISA confirmed MS1 in 2/9 proteins (Clusterin and Serpin A4, P < 0.05), while 3/9 showed differential expression between aAMD and nvAMD (Clusterin, Serpin A4, and TF P < 0.05). Receiver operating characteristic curve calculation identified the area under the curve of 0.82 for clusterin as a biomarker for distinction of AMD.
AH proteomics in AMD patients identified several proteins and functional clusters with altered expression. Further research should confirm if these proteins may serve as biomarkers or therapeutic target for the disease.
年龄相关性黄斑变性（AMD）与视网膜中基因和蛋白质表达的改变有关。我们对 AMD 的房水（AH）蛋白质组进行了特征描述，以深入了解疾病的发病机制并确定潜在的生物标志物。
从年龄和性别匹配的新生血管性 AMD（nvAMD；n=10）患者和对照组（n=10）中收集 AH。将 AH 汇集在一起以创建两个样本（nvAMD 和对照），然后进行基于强度的无标签定量（MS1）。然后进行功能和生物信息学分析。根据 MS1 结果，通过多重 ELISA 对 20 个对照、15 个萎缩性 AMD 和 15 个 nvAMD 的验证集测试了 9 个差异表达蛋白。
MS1 在 AH 中鉴定出 674 种蛋白质。239 种蛋白在 nvAMD 中上调（nvAMD/对照>2，肽标签（PT）>2），86 种蛋白下调（nvAMD/对照<0.5，PT>2）。AMD 中上调蛋白的功能分析表明，血小板脱颗粒（富集分数（ES）：28.1）、内肽酶活性的负调控（ES：18.8）、细胞蛋白代谢过程（ES：11.8）、表皮生长因子样结构域（ES：10.3）、寿司/SCR/CCP（ES：10.1）和补体/凝血级联（ES：9.2）的富集。AMD 蛋白簇上调 3/6（与随机化相比，χ2<0.05）。通过 ELISA 验证在 2/9 种蛋白中证实了 MS1（Clusterin 和 Serpin A4，P<0.05），而 3/9 种蛋白在 aAMD 和 nvAMD 之间显示出差异表达（Clusterin、Serpin A4 和 TF，P<0.05）。接受者操作特征曲线计算确定簇蛋白作为 AMD 鉴别标志物的曲线下面积为 0.82。
AMD 患者的 AH 蛋白质组学鉴定出了几种表达改变的蛋白质和功能簇。进一步的研究应该确认这些蛋白是否可以作为疾病的生物标志物或治疗靶点。
