Expressed Sequence Tag Analysis of Fibrovascular Membranes From Patients With Proliferative Diabetic Retinopathy

Abstract

Purpose: : Preretinal fibrovascular membranes (FVMs) form as a sequela to proliferative diabetic retinopathy (PDR) and can lead to severe decrease of vision. The factors controlling their development and progression have not been fully determined. The purpose of this study was to generate a profile of genes expressed in human FVMs.

Methods: : A polymerase chain reaction (PCR)-amplified cDNA library was constructed using the RNAs isolated from FVMs obtained during vitrectomy from patients with PDR. The sequence from the 5' end was obtained for randomly selected clones and used to generate expressed sequence tags (ESTs). Functional annotation was retrieved from Ensemble database and analyzed by FatiGo. The web-based VisANT software was used to identify the molecular networks within the FVMs.

Results: : A total of 2816 ESTs were assembled in 625 nonredundant clusters. Among these, 515 matched the human cDNA database. The 515 clusters were subdivided by functional subsets of genes related to ribosomal activity, oxidative phosphorylation, focal adhesion, cell adhesion, and other functions. FTL and MALAT1 represented the most abundant transcripts in the FVMs. Querying against VisANT database yielded 3117 possible physical relationships to other genes/proteins which included an additional 1329 genes that were not detected in the FVM library.

Conclusions: : The cDNA library constructed from human FVMs will be a valuable source of information that should facilitate a wide range of studies that can establish the molecular mechanisms underlying the development of FVMs.

Keywords: diabetic retinopathy • gene/expression • proliferation