Lynch Anne M, Drolet Daniel W, Trinder Kinsey M, Gupta Shashi, Westacott Matthew J, Janjic Nebojsa, Palestine Alan G, Patnaik Jennifer L, Mathias Marc T, Mandava Naresh, Wagner Brandie D
Department of Ophthalmology, University of Colorado School of Medicine, Aurora, CO, USA.
Division of Ophthalmic Epidemiology, Department of Ophthalmology, University of Colorado School of Medicine, Mail Stop F731, 1675 Aurora Court, Aurora, CO, 80045, USA.
Clin Proteomics. 2025 Sep 1;22(1):32. doi: 10.1186/s12014-025-09555-3.
BACKGROUND/STUDY OBJECTIVES: Age-related macular degeneration (AMD), a degenerative disease of the photoreceptor support system of the macula, is a leading cause of vision loss in individuals over 60 years of age. In this exploratory longitudinal study, we studied VEGF-related proteins and other protein concentrations in the aqueous humor of patients with treatment naïve neovascular AMD (defined as patients with a previously untreated and recently diagnosed advanced neovascular form of AMD (NVAMD) who were eligible for an intra-vitreal administration of an anti-VEGF agent to treat choroidal neovascularization). The objectives of this small pilot study were: (1) To determine levels of VEGF-related proteins in the aqueous humor of treatment naïve NVAMD patients compared with control patients, (2) To determine whether levels of VEGF-related proteins change over time with anti-VEGF injections in NVAMD patients, (3) To put these differences into perspective relative to all protein targets and identify other off-target (non-VEGF) proteins that may be related to NVAMD or NVAMD treatment.
We used an aptamer-based proteomic technology to study protein concentrations. Cases had a sample of aqueous collected immediately prior to starting the anti-VEGF intra-vitreal injection and at two follow-up visits. Controls were cataract patients with no AMD. Aqueous was collected at the time of cataract surgery.
Comparison between 9 cases and 11 controls revealed 56 proteins, out of 3,803 targets, with significant differences in baseline levels. After treatment, a decline in aqueous VEGF concentrations was indicated from two aptamer reagents, while a third recorded a significant increase. Interference studies demonstrated that the increase in levels observed for the latter reagent was due to measuring both drug-bound and free VEGF concentrations (total VEGF) while the others measured only free VEGF.
In this exploratory study, 56 proteins were identified that could potentially be linked with NVAMD. In interference studies free aqueous VEGF levels declined while total VEGF levels increased following anti-VEGF treatment. No large off-target effects on the proteome were observed with treatment. We illustrate how the protein interactome can mask or potentially unmask binding epitopes leading to signal changes not necessarily related to the absolute protein level.
背景/研究目的：年龄相关性黄斑变性（AMD）是黄斑区光感受器支持系统的一种退行性疾病，是60岁以上人群视力丧失的主要原因。在这项探索性纵向研究中，我们研究了初治新生血管性AMD患者（定义为先前未经治疗且最近诊断为晚期新生血管性AMD（NVAMD）且有资格接受玻璃体内注射抗VEGF药物治疗脉络膜新生血管的患者）房水中VEGF相关蛋白和其他蛋白的浓度。这项小型试点研究的目的是：（1）确定初治NVAMD患者与对照患者房水中VEGF相关蛋白的水平；（2）确定NVAMD患者抗VEGF注射后VEGF相关蛋白水平是否随时间变化；（3）从所有蛋白质靶点的角度看待这些差异，并识别其他可能与NVAMD或NVAMD治疗相关的非靶向（非VEGF）蛋白。
我们使用基于适体的蛋白质组学技术研究蛋白质浓度。病例在开始抗VEGF玻璃体内注射前及两次随访时采集房水样本。对照组为无AMD的白内障患者，在白内障手术时采集房水。
9例病例与11例对照的比较显示，在3803个靶点中，有56种蛋白质的基线水平存在显著差异。治疗后，两种适体试剂显示房水VEGF浓度下降，而第三种试剂记录到显著增加。干扰研究表明，后一种试剂观察到的水平升高是由于同时测量了药物结合型和游离型VEGF浓度（总VEGF），而其他试剂仅测量游离VEGF。
在这项探索性研究中，鉴定出56种可能与NVAMD相关的蛋白质。干扰研究表明，抗VEGF治疗后游离房水VEGF水平下降，而总VEGF水平升高。治疗未观察到对蛋白质组有大的非靶向效应。我们说明了蛋白质相互作用组如何掩盖或潜在地揭示结合表位，导致信号变化不一定与绝对蛋白质水平相关。
