Fig. 7

Fig. 7

A UMAP integrated sc-RNA sequencing identified 15 typical retinal cell clusters. B Expression of ALDH3B1, S1PR5, and FSP1 was primarily observed in microglia, T/NK cells, and fibroblasts, respectively. C–E Integrated RNA-seq data showed gene expression variations in retinal diseases with eovascularization, including AMD (n = 55/14/35/14), BRVO (n = 3/3), and DR (n = 10/27). The expression level of three examined genes were not altered in AMD and BRVO. But in PDR patients, S1PR5 expression was identified to be significantly increased. F–H KEGG pathway enrichment analysis of differential genes highlighted significant changes in pathway categories. I GSEA analysis of transcriptome revealed significant alterations in sphingolipid metabolism and related pathways in PDR retina compared to healthy retina. J GSEA of S1PR5 related genes showed similar changes to those observed in PDR.(K) PCC analysis indicated an expression correlation between S1PR5 expression and ferroptosis-related genes in PDR samples. UMAP, Uniform Manifold Approximation and Projection; sc-RNA, single cell RNA sequence; AMD, age-related macular degeneration; CNV, choroidal neovascularization; BRVO, branch retinal vein occlusion; DR, diabetic retinopathy; PDR, proliferative diabetic retinopathy; KEGG, Kyoto Encyclopedia of Genes and Genomes; GSEA, Gene set enrichment analysis; PCC, Pearson correlation coefficient. Pathway categories were annotated using the KEGG subcategory. S1PR5-related genes were selected with a threshold of R > 0.3 and p < 0.05. For Fig. 7C, E, the center line represents the median, the bounds of the box represent the 25th and 75th percentiles, and the whiskers extend to the largest and smallest values within 1.5 times the interquartile range. Individual points beyond the whiskers are plotted as outliers. Statistical comparisons of gene expression in Fig. 7C–E were performed using Student’s t test. For Fig. 7I, J, GSEA was performed using a permutation-based statistical test. Significance was assessed using the NES and the FDR q-value to account for multiple hypothesis testing. Statistical significance in Fig. 7k was calculated using a two-sided Pearson correlation. Source data are provided as a Source Data file.