Single cell RNA sequencing (scRNA-Seq) deciphering pathological alterations in streptozotocin-induced diabetic retinas

Diabetic retinopathy (DR) is an irreversible and progressive diabetic complication leading to visual impairment, even blindness. Due to the delicate and complicated structure of the retina, the pathology of DR has not been completely elucidated yet. We constructed a transcriptome atlas of >14,000 single cells from healthy and streptozotocin (STZ)-induced diabetic murine retinas to decipher pathological alterations of DR. We found four stress-inducible genes Cirbp, Rmb3, Mt1 and Mt2 commonly induced in most types of retinal cells. Bipolar cells were little affected both in number and gene expression. There existed two subtypes of Müller cells, the Fos-expressing subtype and the no-marker subtype, and the latter lost more in DR. A large number of genes were deregulated in diabetic vascular endothelial cells (ECs), and the differential expression genes were paired to the pathways functioning in metabolism, shear stress and vascular permeability. These pathways were mapped by more differential expression genes in a subpopulation of ECs specifically presented in diabetic retinas (diabetic retinal ECs, DRECs). Moreover, several inflammation pathways were activated in DRECs, and the most significant one is the IL-17 signaling pathway. According to the EC markers, DRECs were mainly capillary ECs, confirmed by immunofluorescent staining of S100a9, a target gene of the IL-17 signaling pathway. This study deciphered pathological alterations of DR, and provided clues for new potential targets for DR therapy. Overall design: The mouse DR model was established by introperitoneal injection of STZ. 25 weeks later, the mice were then sacrificed and retinal single cell suspensions were prepared. The cell suspensions from 3 healthy or 3 diabetic retinas were pooled together. The labaries for scRNA-Seq were construted by the 10X Chromium Single Cell Platform. The final libraries were sequenced on an Illumina Nova6000 instrument using 150-base-pair paired-end reads. UPDATE [04-16-2025] The processed data file was replaced. The new file GSE178121_counts_matrix.csv includes counts for both Samples GSM5380581 and GSM5380582.