Proteomic analysis of aqueous humor in retinoblastoma patients: a focus on intra‐arterial chemotherapy effects

Abstract

AbstractPurpose Proteomics can be a useful tool to understand how therapies act and affect on the molecular composition of ocular fluids. In this retrospective study we focus on how the aqueous humor (AH) proteic pattern from retinoblastoma (RTB) patients is modified by intra‐arterial(IA)treatment.Methods We enrolled 22 RTB patients(Reese stage VB or ABC group E)undergoing enucleation and 20 normal subjects undergoing cataract surgery(CTR).Five of 22 patients presented with associated secondary glaucoma (SG) whereas 17 had no secondary glaucoma (NSG);8 of 17 NSG received IA Melphalan (CHM). Pools were firstly investigated with Bradford method and SDS‐PAGE and then,analyzed using 2D‐DIGE coupled to mass spectrometer.DIA and BVA were fully automated with 2D DeCyder® and statistical analysis was performed with EDA® module.Significant spots were In‐Gel digested and identified with Maldi‐TOF/TOF.Results According to Bradford method,CTR protein content was not significantly different from CHM.After melphalan treatment, total protein contents decreased about 70% while in respect to all non‐treated patients were significantly different(Tamhane p<0.01).PCA analysis underlines high distance either between CTR pool and SG/NSG pool or CHM and RTB.In particular the analysis of single protein shows that CHM pool is characterized by a high level of Crystallin and pro‐inflammatory proteins and CTR pool is characterized by a high level of Epithelium Growth Factor.Conclusion IA chemotherapy with Melphalan modifies the AH proteic pattern revealing the presence of Crystallin family and pro‐inflammatory proteins. This opens new horizons to further explore local pharmacological effects of new drugs.