Nobl Matthias, Reich Michael, Dacheva Ivanka, Siwy Justyna, Mullen William, Schanstra Joost P, Choi Chul Young, Kopitz Jürgen, Kretz Florian T A, Auffarth Gerd U, Koch Frank, Koss Michael J
Department of Ophthalmology, University of Heidelberg, Germany.
Department of Ophthalmology, University of Heidelberg, Germany; Department of Ophthalmology, University of Freiburg, Germany.
Exp Eye Res. 2016 May;146:107-117. doi: 10.1016/j.exer.2016.01.001. Epub 2016 Jan 6.
Neovascular age-related macular degeneration (nAMD) has been described as a predominantly inflammatory and proangiogenic retino-choroidal disease. Vitreous humor (VH) is the adjacent and accessible compartment which, due to the vicinity to the retina, might best represent changes of protein-based mediators of nAMD. The aim of this clinical-experimental study was to analyze the nAMD associated VH proteome of previously untreated patients whilst taking different groups of nAMD into account, based on their clinical presentation (clinical diagnosis groups). Electrophoresis coupled online to mass spectrometry (CE-MS) as well as liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) were used to analyze VH of 108 nAMD patients and 24 controls with idiopathic floaters. A total of 101 different proteins with at least two unique peptides could be identified. Using a stringent statistical analysis with implementation of the closed test principle, we were able to identify four proteins that may be involved in the pathophysiology of nAMD: Clusterin, opticin, pigment epithelium-derived factor and prostaglandin-H2 d-isomerase. Using independent samples, ROC-Area under the curve was determined proving the validity of the results: Clusterin 0.747, opticin 0.656, pigment epithelium-derived factor 0.514, prostaglandin-H2 d-isomerase 0.712. In addition, validation through ELISA measurements was performed. The identified proteins may serve as potential biomarkers or even targets of therapy for nAMD.
新生血管性年龄相关性黄斑变性（nAMD）被描述为一种主要是炎症性和促血管生成的视网膜脉络膜疾病。玻璃体液（VH）是相邻且易于获取的腔室，由于其靠近视网膜，可能最能代表nAMD基于蛋白质的介质变化。本临床实验研究的目的是分析未经治疗的患者的nAMD相关玻璃体液蛋白质组，同时根据临床表现（临床诊断组）考虑不同类型的nAMD。采用在线电泳联用质谱（CE-MS）以及液相色谱联用串联质谱（LC-MS/MS）分析了108例nAMD患者和24例患有特发性飞蚊症的对照者的玻璃体液。总共鉴定出101种不同的蛋白质，每种蛋白质至少有两个独特的肽段。通过实施封闭检验原则进行严格的统计分析，我们能够鉴定出四种可能参与nAMD病理生理学的蛋白质：簇集蛋白、视蛋白、色素上皮衍生因子和前列腺素-H2 d-异构酶。使用独立样本，确定了曲线下面积（ROC-AUC），证明了结果的有效性：簇集蛋白为0.747，视蛋白为0.656，色素上皮衍生因子为0.514，前列腺素-H2 d-异构酶为0.712。此外，还通过酶联免疫吸附测定（ELISA）进行了验证。所鉴定的蛋白质可能作为nAMD的潜在生物标志物甚至治疗靶点。
