β-Cell pre-mir-21 induces dysfunction and loss of cellular identity by targeting transforming growth factor beta 2 (Tgfb2) and Smad family member 2 (Smad2) mRNAs
Objective: β-cell microRNA-21 (miR-21) is increased by islet inflammatory stress but it decreases glucose-stimulated insulin secretion (GSIS). Thus, we sought to define the effects of miR-21 on β-cell function using in vitro and in vivo systems. Methods: We developed a tetracycline-on system of pre-...
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doaj-d4546e0f597946beb741de5f5f20e4002021-08-10T04:04:55ZengElsevierMolecular Metabolism2212-87782021-11-0153101289β-Cell pre-mir-21 induces dysfunction and loss of cellular identity by targeting transforming growth factor beta 2 (Tgfb2) and Smad family member 2 (Smad2) mRNAsSara Ibrahim0Macey Johnson1Clarissa Hernandez Stephens2Jerry Xu3Rachel Moore4Andrea Mariani5Christopher Contreras6Farooq Syed7Raghavendra G. Mirmira8Ryan M. Anderson9Emily K. Sims10Department of Biochemistry and Molecular Biology, USA; Center for Diabetes and Metabolic Diseases, USACenter for Diabetes and Metabolic Diseases, USACenter for Diabetes and Metabolic Diseases, USA; Department of Pediatrics, USACenter for Diabetes and Metabolic Diseases, USA; Department of Pediatrics, USA; Wells Center for Pediatric Research, USACenter for Diabetes and Metabolic Diseases, USA; Department of Pediatrics, USACenter for Diabetes and Metabolic Diseases, USA; Department of Pediatrics, USACenter for Diabetes and Metabolic Diseases, USA; Department of Pediatrics, USACenter for Diabetes and Metabolic Diseases, USA; Wells Center for Pediatric Research, USAKovler Diabetes Center and the Department of Medicine, The University of Chicago, Chicago, IL, 60637, USAKovler Diabetes Center and the Department of Medicine, The University of Chicago, Chicago, IL, 60637, USA; Corresponding authorCenter for Diabetes and Metabolic Diseases, USA; Department of Pediatrics, USA; Wells Center for Pediatric Research, USA; Corresponding author. Indiana University School of Medicine, 635 Barnhill Drive MS 2031E, Indianapolis, IN, USA. Fax: +317 944 3882.Objective: β-cell microRNA-21 (miR-21) is increased by islet inflammatory stress but it decreases glucose-stimulated insulin secretion (GSIS). Thus, we sought to define the effects of miR-21 on β-cell function using in vitro and in vivo systems. Methods: We developed a tetracycline-on system of pre-miR-21 induction in clonal β-cells and human islets, along with transgenic zebrafish and mouse models of β-cell-specific pre-miR-21 overexpression. Results: β-cell miR-21 induction markedly reduced GSIS and led to reductions in transcription factors associated with β-cell identity and increased markers of dedifferentiation, which led us to hypothesize that miR-21 induces β-cell dysfunction by loss of cell identity. In silico analysis identified transforming growth factor-beta 2 (Tgfb2) and Smad family member 2 (Smad2) mRNAs as predicted miR-21 targets associated with the maintenance of β-cell identity. Tgfb2 and Smad2 were confirmed as direct miR-21 targets through RT-PCR, immunoblot, pulldown, and luciferase assays. In vivo zebrafish and mouse models exhibited glucose intolerance, decreased peak GSIS, decreased expression of β-cell identity markers, increased insulin and glucagon co-staining cells, and reduced Tgfb2 and Smad2 expression. Conclusions: These findings implicate miR-21-mediated reduction of mRNAs specifying β-cell identity as a contributor to β-cell dysfunction by the loss of cellular differentiation.http://www.sciencedirect.com/science/article/pii/S2212877821001344β-cellIsletmicroRNA 21DedifferentiationIdentityβ-cell dysfunction |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Sara Ibrahim Macey Johnson Clarissa Hernandez Stephens Jerry Xu Rachel Moore Andrea Mariani Christopher Contreras Farooq Syed Raghavendra G. Mirmira Ryan M. Anderson Emily K. Sims |
spellingShingle |
Sara Ibrahim Macey Johnson Clarissa Hernandez Stephens Jerry Xu Rachel Moore Andrea Mariani Christopher Contreras Farooq Syed Raghavendra G. Mirmira Ryan M. Anderson Emily K. Sims β-Cell pre-mir-21 induces dysfunction and loss of cellular identity by targeting transforming growth factor beta 2 (Tgfb2) and Smad family member 2 (Smad2) mRNAs Molecular Metabolism β-cell Islet microRNA 21 Dedifferentiation Identity β-cell dysfunction |
author_facet |
Sara Ibrahim Macey Johnson Clarissa Hernandez Stephens Jerry Xu Rachel Moore Andrea Mariani Christopher Contreras Farooq Syed Raghavendra G. Mirmira Ryan M. Anderson Emily K. Sims |
author_sort |
Sara Ibrahim |
title |
β-Cell pre-mir-21 induces dysfunction and loss of cellular identity by targeting transforming growth factor beta 2 (Tgfb2) and Smad family member 2 (Smad2) mRNAs |
title_short |
β-Cell pre-mir-21 induces dysfunction and loss of cellular identity by targeting transforming growth factor beta 2 (Tgfb2) and Smad family member 2 (Smad2) mRNAs |
title_full |
β-Cell pre-mir-21 induces dysfunction and loss of cellular identity by targeting transforming growth factor beta 2 (Tgfb2) and Smad family member 2 (Smad2) mRNAs |
title_fullStr |
β-Cell pre-mir-21 induces dysfunction and loss of cellular identity by targeting transforming growth factor beta 2 (Tgfb2) and Smad family member 2 (Smad2) mRNAs |
title_full_unstemmed |
β-Cell pre-mir-21 induces dysfunction and loss of cellular identity by targeting transforming growth factor beta 2 (Tgfb2) and Smad family member 2 (Smad2) mRNAs |
title_sort |
β-cell pre-mir-21 induces dysfunction and loss of cellular identity by targeting transforming growth factor beta 2 (tgfb2) and smad family member 2 (smad2) mrnas |
publisher |
Elsevier |
series |
Molecular Metabolism |
issn |
2212-8778 |
publishDate |
2021-11-01 |
description |
Objective: β-cell microRNA-21 (miR-21) is increased by islet inflammatory stress but it decreases glucose-stimulated insulin secretion (GSIS). Thus, we sought to define the effects of miR-21 on β-cell function using in vitro and in vivo systems. Methods: We developed a tetracycline-on system of pre-miR-21 induction in clonal β-cells and human islets, along with transgenic zebrafish and mouse models of β-cell-specific pre-miR-21 overexpression. Results: β-cell miR-21 induction markedly reduced GSIS and led to reductions in transcription factors associated with β-cell identity and increased markers of dedifferentiation, which led us to hypothesize that miR-21 induces β-cell dysfunction by loss of cell identity. In silico analysis identified transforming growth factor-beta 2 (Tgfb2) and Smad family member 2 (Smad2) mRNAs as predicted miR-21 targets associated with the maintenance of β-cell identity. Tgfb2 and Smad2 were confirmed as direct miR-21 targets through RT-PCR, immunoblot, pulldown, and luciferase assays. In vivo zebrafish and mouse models exhibited glucose intolerance, decreased peak GSIS, decreased expression of β-cell identity markers, increased insulin and glucagon co-staining cells, and reduced Tgfb2 and Smad2 expression. Conclusions: These findings implicate miR-21-mediated reduction of mRNAs specifying β-cell identity as a contributor to β-cell dysfunction by the loss of cellular differentiation. |
topic |
β-cell Islet microRNA 21 Dedifferentiation Identity β-cell dysfunction |
url |
http://www.sciencedirect.com/science/article/pii/S2212877821001344 |
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