Glucocorticoid induces human beta cell dysfunction by involving riborepressor GAS5 LincRNA

Jonathan L.S Esguerra,Jones K Ofori,Mototsugu Nagao,Yuki Shuto,Alexandros Karagiannopoulos,Joao Fadista,Hitoshi Sugihara,Leif Groop,Lena Eliasson

doi:10.1016/j.molmet.2019.12.012

Jonathan L.S Esguerra, Jones K Ofori + Show 7 more

Open Access

https://doi.org/10.1016/j.molmet.2019.12.012

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Abstract

ObjectiveA 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. The significance of non-coding RNAs in various cellular processes is emerging. In this study, we aimed to show the direct negative impact of GC on beta cell function and elucidate the role of riborepressor GAS5 lincRNA in the GC signaling pathway in human pancreatic beta cells. MethodsPatients undergoing two weeks of 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. The GAS5 level was modulated using anti-sense LNA gapmeR or 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. ResultsWe 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 exocytotic protein SYT13 were observed. The expression of the glucocorticoid receptor was decreased, while that of serum and glucocorticoid-regulated kinase 1 (SGK1) 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. ConclusionsThe 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 GAS5 lincRNA, a potentially novel therapeutic target to counter GC-mediated beta cell dysfunction.

Highlights

Glucocorticoids (GCs) in various forms are highly potent steroid hormones in the frontline of various clinical therapy procedures
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 growth arrest-specific 5 (GAS5) long intergenic non-coding RNA (lincRNA), a potentially novel therapeutic target to counter GC-mediated beta cell dysfunction
Glucocorticoid treatment reduces insulin secretion at high glucose concentrations and GAS5 expression in human islets and beta cells The contribution of GC-mediated beta cell dysfunction to causing hyperglycemia was previously shown in vivo in which healthy individuals were subjected to acute or two-week prednisolone treatment resulting in reduced insulin secretion [15]

Summary

Introduction

Glucocorticoids (GCs) in various forms (hydrocortisone, dexamethasone, prednisolone, and prednisone) are highly potent steroid hormones in the frontline of various clinical therapy procedures. They are the most widely prescribed drugs used for treating allergic disorders, inflammatory and autoimmune diseases, some forms of malignancies, and suppressing immune response following organ transplantation [1,2]. Despite the efficacy of GC therapy in treating human diseases, metabolic side effects have been acknowledged, of which steroidinduced diabetes mellitus (DM) is the most recognized [3]. Despite ample evidence of the direct effects of GCs in rodent beta cells [9e11], studies on the molecular basis of GC-induced pancreatic beta cell dysfunction in human beta cells are lacking. An emerging complexity in gene regulation involves non-protein coding functional RNA molecules that can significantly influence diverse

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