Glucocorticoid induces human beta cell dysfunction by involving riborepressor GAS5 LincRNA

Abstract Objective A widely recognized metabolic side-effect of glucocorticoid (GC) therapy is steroid-induced diabetes mellitus (DM). However, studies on the molecular basis of GC-induced pancreatic beta cell dysfunction in human beta cells are lacking. Meanwhile, the significance of non-coding RNAs in various cellular processes is emerging. Here we aimed to show the direct negative impact of GC on beta cell function and elucidate the role of the riborepressor GAS5 lincRNA in GC signaling pathway in human pancreatic beta cells. Methods Patients undergoing two weeks high-dose prednisolone therapy were monitored for c-peptide levels. Human pancreatic islets and the human beta cell line, EndoC-βH1, were incubated in pharmacological concentrations of dexamethasone. GAS5 level was modulated using anti-sense LNA gapmer or using short oligonucleotides with GAS5 HREM (hormone response element motif). Immunoblotting and/or real-time PCR were used to assess changes in protein and RNA expression, respectively. Functional characterization included glucose-stimulated insulin secretion and apoptosis assays. Correlation analysis was performed on RNAseq data of human pancreatic islets. Results We found reduced c-peptide levels in patients undergoing high dose GC therapy. Human islets and the human beta cell line, EndoC-βH1, exposed to GC exhibited reduced insulin secretion and increased apoptosis. Concomitantly, reduced expression of important beta cell transcription factors, PDX1 and NKX6.1, as well as the exocytotic protein, SYT13 was observed. Furthermore, the expression of the glucocorticoid receptor was decreased, while SGK1 (serum and glucocorticoid-regulated kinase 1) was elevated. The expression of these genes was found to significantly correlate with GAS5 in human islet transcriptomics data. Increasing GAS5 levels using GAS5 HREM alleviated the inhibitory effects of dexamethasone on insulin secretion. Conclusions The direct adverse effect of glucocorticoid in human beta cell function is mediated via important beta cell proteins and components of the GC-signaling pathway in an intricate interplay with the GAS5 lincRNA, a potentially novel therapeutic target to counter GC-mediated beta cell dysfunction.