Proteomic analysis of aqueous humor in retinoblastoma: Final results

Abstract

Purpose Proteomics in ocular fluids is gaining rapidly relevance.Since 2009 we work on the analysis of proteic composition of the aqueous humor (AH) from retinoblastoma (RTB) patients and,in this work,we use the 2D‐DIGE coupled to mass spectrometer to find new RTB markers. Methods We enrolled 22 RTB patients(Reese‐Ellsworth stage V or ABC classification group E)undergoing enucleation and 20 normal subjects undergoing cataract surgery(CTR).Five of 22 patients presented with associated secondary glaucoma whereas 17 had no secondary glaucoma;8 of 17 patients with no secondary glaucoma received chemotherapeutical treatment with melphalan.Pools were 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 PCA analysis of the data underlines high distance between CTR pool and RTB pools.In particular the analysis of single protein shows that all the RTB pools are characterized by a high level of Crystallin and pro‐inflammatory proteins and CTR pool are characterized by a high level of Epithelium Growth Factor. Conclusion This study is the last step for the clarification of AH protein pattern in patients with RTB.Although we learn from literature that the presence of inflammatory processes is widely documented in late stages of pathology,however the presence of crystalline proteins could be either a new marker or an active part of the late stage tumor‐induced panorama in RTB AH samples.Due to this,we think that several molecular biology studies are necessary to better understand involvement of this protein family in RTB pathological pathway.