Multi-modal proteomic target discovery and orthogonol confirmation of preclinical diabetic retinopathy drug development biomarkers

The aim of this study was to identify proteomic alterations associated with functional dysregulation of the retina with diabetes that could eventually be used as surrogate endpoints in preclinical drug testing studies. A multi-modal approach of antibody (Luminex)-, electrophoresis (2-DIGE)-, and LC-MS (iTRAQ)-based quantitation methods was used to provide broad coverage of the retinal proteome. Transcriptional profiling through microarray analysis was also included to increase coverage and provide insight into potential regulation of protein expression changes at the mRNA level. The different technologies proved complementary, with limited coverage overlap between methods.本研究旨在识别与糖尿病性视网膜功能失调相关的蛋白质组学改变（proteomic alterations），此类改变最终可作为临床前药物试验研究中的替代终点（surrogate endpoints）。本研究采用多模态研究策略，整合基于抗体（Luminex）、电泳（2-DIGE，双向荧光差异凝胶电泳）以及液相色谱-质谱联用（iTRAQ，同位素标记相对和绝对定量）的定量检测方法，以实现视网膜蛋白质组（retinal proteome）的广覆盖度分析。此外，本研究还纳入基因芯片分析（microarray analysis）以开展转录组谱分析（transcriptional profiling），以此提升覆盖度并阐明蛋白质表达变化在mRNA层面的潜在调控机制。实验结果表明，不同检测技术间具有互补性，各方法的覆盖度重叠范围有限。