RNA-Binding Proteins and Alternative Splicing Genes Are Coregulated in Human Retinal Endothelial Cells Treated with High Glucose.

Hongran Zhao,Tianran Chen,Bozhao Wang,Hui Kong,Yan Cui,Sihui Wu,Hiroshi Okamoto

doi:10.1155/2022/7680513

Abstract

To explore the relevant RNA-binding proteins (RBPs) and alternative splicing events (ASEs) in diabetic retinopathy (DR). We devised a comprehensive work to integrate analyses of the differentially expressed genes, including differential RBPs, and variable splicing characteristics related to DR in human retinal endothelial cells induced by low glucose and high glucose in dataset GSE117238. A total of 2320 differentially expressed genes (DEGs) were identified, including 1228 upregulated genes and 1092 downregulated genes. Further analysis screened out 232 RBP genes, and 42 AS genes overlapped DEGs. We selected high expression and consistency six RBP genes (FUS, HNRNPA2B1, CANX, EIF1, CALR, and POLR2A) for coexpression analysis. Through analysis, we found eight RASGs (MDM2, GOLGA2P7, NFE2L1, KDM4A, FAM111A, CIRBP, IDH1, and MCM7) that could be regulated by RBP. The coexpression network was conducted to further elucidate the regulatory and interaction relationship between RBPs and AS. Apoptotic progress, protein phosphorylation, and NF-kappaB cascade revealed by the functional enrichment analysis of RASGs regulated by RBPs were closely related to diabetic retinopathy. Furthermore, the expression of differentially expressed RBPs was validated by qRT-PCR in mouse retinal microvascular endothelial cells and retinas from the streptozotocin mouse model. The results showed that Fus, Hnrnpa2b1, Canx, Calr, and Polr2a were remarkedly difference in high-glucose-treated retinal microvascular endothelial cells and Fus, Hnrnpa2b1, Canx, and Calr were remarkedly difference in retinas from streptozotocin-induced diabetic mice compared to control. The regulatory network between identified RBPs and RASGs suggests the presence of several signaling pathways possibly involved in the pathogenesis of DR. The verified RBPs should be further addressed by future studies investigating associations between RBPs and the downstream of AS, as they could serve as potential biomarkers and targets for DR.

Highlights

Diabetic retinopathy (DR) is a common and specific microvascular complication of diabetes mellitus and a leading cause of vision impairment and blindness in the workingage population [1]
Genes Are Differentially Expressed in Human Retinal Endothelial Cells (HRECs) Treated with Low Glucose (LG) and High Glucose (HG)
We found eight regulated alternative splicing genes (RASGs) (MDM2, GOLGA2P7, NFE2L1, KDM4A, FAM111A, CIRBP, IDH1, and MCM7) that could be regulated by RNA-binding proteins (RBPs) (Figures 3(d) and 3(f))

Summary

Introduction

Diabetic retinopathy (DR) is a common and specific microvascular complication of diabetes mellitus and a leading cause of vision impairment and blindness in the workingage population [1]. It has been estimated that 693 million people will have diabetes by 2045, 35% of whom will have DR. Severe degrees of DR have been associated with impaired quality of life, reduced physical, emotional, and social well-being, as well as a serious family burden [2]. The current treatments for DR are applicable only at advanced stages and often associated with poor prognosis, while early diagnosis and preventative measures are of great importance to prevent DR. Further understanding of the key mechanism that could lead to early recognition of DR is vital for preventing the degradation of visual acuity and developing new, more effective treatments.

Results

Discussion

Conclusion