RNAseq of Mineralocorticoid Receptor (MR) overexpression in UVB-induced ocular rosacea model on meibomian glands of rats
The aim of this study was to analyse the transcriptional regulation of UVB-induced ocular rosacea model on the meibomian glands of rats that overexpress the human NRC32 gene coding for mineralocorticoid receptor (P1.hMR) and age- and sex- matched wild type (WT) littlemates. 10-12 weeks old female Wild-Type (WT) and P1.hMR rats were subjected to 5-days UVB irrdiation on eyelids (UVB_WT: n = 3, UVB_P1.hMR: n = 5), together with age- and sex- matched rats without UVB-induced model (Control_WT: n = 4, Control_P1.hMR: n = 5), they were euthanized and the meibomian glands were dissected 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 and RNA integrity numbers (RIN) ranging from 8.8 to 9.3 was accepted for library generation. Quality of raw data was assessed using FastQC. Poor quality sequences and adapters were trimmed or removed with fastp, using default parameters, to retain only good quality paired reads. Illumina DRAGEN bio-IT Plateform (v3.8.4) was used for mapping on rn7 reference genome. Library orientation, library composition and coverage along transcripts were checked with Picard tools. Differential analysis has been also conducted with DESeq2. Multiple hypothesis adjusted p-values were calculated with the Benjamini-Hochberg procedure to control FDR. FDR threshold was set at 0.05, log2 fold-change threshold was set at 0.5. We identified genes differentially regulated in UVB-treated WT as compared to Control WT, and genes differentially regulated in UVB-treated P1.hMR rats as compared to Control P1.hMR. Overall design: UVB-induced gene regulation on meibomian glands mRNA profiles of rats that overexpressed the human NR3C2 gene coding for mineralocorticoid receptor.