RNAseq of the peripheral cornea and limboscleral tissues of rats treated with systemic spironolactone in a model of corneal neovascularization
The aim of this study was to analyse the transcriptional regulation in the corneal and limbal tissues of rats treated with spironolactone in a model of limbal stem cell deficiency (LSCD). CNV was induced by total corneal de-epithelialization and limbal resection. Rats were injected subcutaneously with spironolactone (25mg/kg/day) diluted in 90% olive oil and 10% DMSO  or vehicle (90% olive oil and 10% DMSO) for 14 days. On day 3 (early inflammatory stage) and day 7 (angiogenic stage), rats were euthanized and the peripheral cornea and the limboscleral tissues were exised for RNA-sequencing. Total RNA samples were sequenced at the iGenSeq transcriptomic platform of the Brain and Spine Institute (ICM, Paris, France). RNA quality was checked by capillary electrophoresis (Agilent 2100 Bioanalyzer system) and RNA with integrity numbers (RIN) ranging from 7.8 to 8.2 was accepted for library generation. Quality of raw data has been evaluated with FastQC. Illumina DRAGEN bio-IT Platform (v3,8,4) was used to align reads on reference genome rd6 and quantification with genecode vM25 annotation gtf. Library orientation, library composition and coverage along transcripts were checked with Picard tools. differential analysis has been also conducted with edgeR. Multiple hypothesis adjusted p-values were calculated with the Benjamini-Hochberg procedure to control FDR. We identified genes differentially regulated by spironolactone vs. vehicle on day 3 and day 7. Overall design: Corneal and limboscleral mRNA profiles of rats treated with systemic spironolactone in a rat model of limbal stem cell deficiency