Abstract
Purpose :
We have previously shown that the treatment with a galectin-3 inhibitor, GAL-200-10, reduces corneal as well subretinal fibrosis. To further attest to the efficacy of galectin-3 inhibition in the reduction of fibrosis, and to shed light on the mechanism by which galectin-3 inhibition reduces fibrosis, the goal of the current study was to identify differentially expressed genes in alkali-burned corneas treated with the galectin-3 inhibitor.
Methods :
Corneal fibrosis was induced by alkali-burn injury in C57B/6 mice. Vehicle alone (10µl of 5% PEG400/0.75% HEC/0.5% HPMC 606/ 0.1% Poloxamer 407) or GAL-200-10 (10µl of 10mg/ml in vehicle) were topically applied to the eye twice per day from day 1 until day 14. Corneal opacity was scored by slit lamp examination at day 7 and 14 post-injury. On day 14, corneas were harvested, RNA was isolated and processed: (i) for RNAseq transcriptome using the Illumina Hiseq 2500 sequencing system and (ii) for Nanostring analysis using a mouse fibrosis panel. Differentially expressed genes were used to perform pathway analyses using Ingenuity pathway Analysis (IPA) and nCounter gene expression platform analysis.
Results :
RNA-Seq analysis revealed that in the GAL-200-10-treated group, seven genes were upregulated, and 242 genes were downregulated (cutoff > 50 reads and P value cutoff of 0.05). Most differentially expressed genes were related to tissue fibrosis such as cellular movement, and inflammatory response. NanoString analysis using a mouse fibrosis panel confirmed our RNAseq analysis and showed that the majority of the fibrosis-related genes were downregulated in the Gal3 inhibitor-treated group compared to the vehicle-treated group (cutoff > 20 reads and P value cutoff of 0.05).
Conclusions :
Our data demonstrating the downregulation of numerous fibrosis-related genes in the corneas treated with GAL-200-10, further attest to the efficacy of galectin-3 inhibition in the reduction of fibrosis and, lead us to conclude that the galectin-3 inhibitor, GAL-200-10 is an excellent candidate for developing a new drug to halt the progression of fibrosis after injury.
This is a 2020 ARVO Annual Meeting abstract.