Department of Ophthalmology, Seoul Metropolitan Government, Seoul National University Boramae Medical Center, Seoul, Republic of Korea.
J Proteome Res. 2012 Aug 3;11(8):4034-43. doi: 10.1021/pr300080s. Epub 2012 Jul 13.
Age-related macular degeneration (AMD) can lead to irreversible central vision loss in the elderly. Although large number of growth factor pathways, including the vascular endothelial growth factor (VEGF), has been implicated in the pathogenesis of AMD, no study has directly assessed the whole proteomic composition in the aqueous humor (AH) among AMD patients. The AH contains proteins secreted from the anterior segment tissue, and these proteins may play an important role in the pathogenesis of AMD. Thus, comparisons between the AH proteomic profiles of AMD patients and non-AMD controls may lead to the verification of novel pathogenic proteins useful as potential clinical biomarkers. In this study, we used discovery-based proteomics and Multiple Reaction Monitoring Mass Spectrometry (MRM-MS) to analyze AH from AMD patients and AH from controls who underwent cataract surgery. A total of 154 proteins with at least two unique peptides were identified in the AH. Of these 154 proteins identified by discovery-based proteomics, 10 AH proteins were novel identifications. The protein composition in the AH was different between AMD patients and non-AMD controls. Subsequently, a systematic MRM-MS assay was performed in seven highly abundant differentially expressed proteins from these groups. Differential expression of three proteins was observed in the AH of AMD patients compared with that of cataract controls (p<0.0312). Elucidation of the aqueous proteome will establish a foundation for protein function analysis and identify differentially expressed markers associated with AMD. This study demonstrates that integrated proteomic technologies can yield novel biomarkers to detect exudative AMD.
年龄相关性黄斑变性（AMD）可导致老年人视力不可逆转的中心损失。尽管大量的生长因子途径，包括血管内皮生长因子（VEGF），已被牵连在 AMD 的发病机制中，但没有研究直接评估 AMD 患者房水中的整体蛋白质组组成。房水包含从前节组织分泌的蛋白质，这些蛋白质可能在 AMD 的发病机制中发挥重要作用。因此，AMD 患者和非 AMD 对照组房水蛋白质组谱之间的比较可能会验证新的致病蛋白，这些蛋白可用作潜在的临床生物标志物。在这项研究中，我们使用基于发现的蛋白质组学和多重反应监测质谱法（MRM-MS）分析了接受白内障手术的 AMD 患者和非 AMD 对照组的房水。在房水中共鉴定出 154 种至少有两种独特肽的蛋白质。在基于发现的蛋白质组学鉴定的 154 种蛋白质中，有 10 种房水蛋白质是新的鉴定结果。AMD 患者和非 AMD 对照组的房水蛋白质组成不同。随后，对来自这些组的七个高度丰富的差异表达蛋白进行了系统的 MRM-MS 检测。与白内障对照组相比，AMD 患者房水中有三种蛋白质的表达差异（p<0.0312）。阐明房水蛋白质组将为蛋白质功能分析建立基础，并确定与 AMD 相关的差异表达标志物。本研究表明，综合蛋白质组学技术可以产生新的生物标志物来检测渗出性 AMD。
