Proteomic changes in aqueous humor, anterior capsule and crystalline lens in diabetic and age-related cataract.

Abstract

Purpose : The comprehensive investigation of cataract formation is essential in order to identify non-invasive therapeutic targets. Proteomics comprises the most precious method for deciphering cataract pathophysiology. The aim of this study is to detect proteome changes and corresponding signaling pathways involved in the development of diabetic cataract (DC) and age-related cataract (ARC).

Methods : This study was conducted according to the ethical principles of medical research involving human subjects according to the Declaration of Helsinki. Three sample groups, e.g. the aqueous humor, the anterior capsule and the content of the phaco cassette, were collected during cataract surgery. The samples were collected from 11 patients with diabetes mellitus, younger than 65 years old, and from 12 patients without diabetes mellitus, older than 75 years old. The samples were prepared based on Sp3 protocol (Single-Pot, Solid-Phase-enhanced, Sample-Preparation). The recognition and quantification of proteins was performed with liquid chromatography online with tandem mass spectrometry. Data analysis was performed using the DIA-NN software for identification and quantification of peptides and proteins. Perseus software was used for statistical analysis (two samples t-test) and data visualization.

Results : The study has generated a very rich atlas of the eye proteome. Non-canonical Wnt receptor signaling pathway was significantly upregulated in anterior capsule and phaco cassette samples of ARC. Glycosphingolipid and glycosaminoglycan biosynthesis pathways were significantly upregulated in all three sample types of ARC. Pathways involved in microtubule cytoskeleton organization were upregulated in all three sample types of DC. Structural constituent of the eye lens pathway was upregulated in anterior capsule and phaco cassette samples of DC. Proteasome core complex was upregulated in aqueous humor samples of DC.

Conclusions : Outstanding proteomic profiles of the three sample types have been generated using DIA proteomics. Grouping of the patients allows deciphering of numerous deregulated pathways correlated with cataract. The generated data sets may be useful in finding protein druggable targets against cataract.

This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.