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Glucocorticoid induces human beta cell dysfunction by involving riborepressor GAS5 LincRNA

Esguerra, Jonathan L.S. LU orcid ; Ofori, Jones K. LU ; Nagao, Mototsugu LU ; Shuto, Yuki ; Karagiannopoulos, Alexandros LU orcid ; Fadista, Joao LU ; Sugihara, Hitoshi ; Groop, Leif LU and Eliasson, Lena LU orcid (2020) In Molecular Metabolism 32. p.160-167
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. 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. Methods: Patients 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... (More)

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. 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. Methods: Patients 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. 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 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. 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 GAS5 lincRNA, a potentially novel therapeutic target to counter GC-mediated beta cell dysfunction.

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author
; ; ; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Beta cells, Glucocorticoid, Insulin secretion, Long intergenic non-coding RNA, Pancreatic islets, Type-2 diabetes mellitus
in
Molecular Metabolism
volume
32
pages
8 pages
publisher
Elsevier
external identifiers
  • scopus:85078576325
  • pmid:32029226
ISSN
2212-8778
DOI
10.1016/j.molmet.2019.12.012
language
English
LU publication?
yes
id
aa3fa81d-c161-4ffa-a643-e029531dab61
date added to LUP
2020-02-09 21:33:12
date last changed
2024-06-13 11:10:51
@article{aa3fa81d-c161-4ffa-a643-e029531dab61,
  abstract     = {{<p>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. 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. Methods: Patients 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. 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 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. 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 GAS5 lincRNA, a potentially novel therapeutic target to counter GC-mediated beta cell dysfunction.</p>}},
  author       = {{Esguerra, Jonathan L.S. and Ofori, Jones K. and Nagao, Mototsugu and Shuto, Yuki and Karagiannopoulos, Alexandros and Fadista, Joao and Sugihara, Hitoshi and Groop, Leif and Eliasson, Lena}},
  issn         = {{2212-8778}},
  keywords     = {{Beta cells; Glucocorticoid; Insulin secretion; Long intergenic non-coding RNA; Pancreatic islets; Type-2 diabetes mellitus}},
  language     = {{eng}},
  month        = {{02}},
  pages        = {{160--167}},
  publisher    = {{Elsevier}},
  series       = {{Molecular Metabolism}},
  title        = {{Glucocorticoid induces human beta cell dysfunction by involving riborepressor GAS5 LincRNA}},
  url          = {{http://dx.doi.org/10.1016/j.molmet.2019.12.012}},
  doi          = {{10.1016/j.molmet.2019.12.012}},
  volume       = {{32}},
  year         = {{2020}},
}