Abstract
Aims
MicroRNAs are a class of negative regulators of gene expression, which have been shown to be involved in the development of endocrine pancreas and in the regulation of insulin secretion. Since type 2 diabetes (T2D) is characterized by beta cell dysfunction, we aimed at evaluating expression levels of miR-124a and miR-375, both involved in the control of beta cell function, in human pancreatic islets obtained from T2D and from age-matched non-diabetic organ donors.
Methods
We analyzed miR-124a and miR-375 expression by real-time qRT-PCR in human pancreatic islets and evaluated the potential role of miR-124a by overexpressing or silencing such miRNA in MIN6 pseudoislets.
Results
We identified a major miR-124a hyperexpression in T2D human pancreatic islets with no differential expression of miR-375. Of note, miR-124a overexpression in MIN6 pseudoislets resulted in an impaired glucose-induced insulin secretion. In addition, miR-124a silencing in MIN6 pseudoislets resulted in increased expression of predicted target genes (Mtpn, Foxa2, Flot2, Akt3, Sirt1 and NeuroD1) involved in beta cell function. For Mtpn and Foxa2, we further demonstrated the actual binding of miR-124a to their 3UTR sequences by luciferase assay.
Conclusions
We uncovered a major hyperexpression of miR-124a in T2D islets, whose silencing resulted in increased expression of target genes of major importance for beta cell function and whose overexpression impaired glucose-stimulated insulin secretion, leading to the hypothesis that an altered miR-124a expression may contribute to beta cell dysfunction in type 2 diabetes.
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Acknowledgments
This work was supported by grants from the European Union [Collaborative Projects NAIMIT and PEVNET in the Framework Program 7 (FP7)], from the Italian Ministry of Research and from the Tuscany Region. The editorial help of Maddalena Prencipe has been greatly appreciated.
Conflict of interest
Guido Sebastiani, Agnese Po, Evelina Miele, Giuliana Ventriglia, Elena Ceccarelli, Marco Bugliani, Lorella Marselli, Piero Marchetti, Alberto Gulino, Elisabetta Ferretti and Francesco Dotta declare that they have no conflict of interest.
Ethical standard
All human studies have been reviewed by the appropriate ethics committee (Comitato Etico per la Sperimentazione dell’Azienda Ospedaliera Universitaria di Pisa) and have therefore been performed in accordance with the ethical standards.
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All procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and with the Helsinki Declaration of 1975, as revised in 2008 (5).
Informed consent disclosure
Informed consent was obtained from all patients for being included in the study.
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Managed by Massimo Federici.
Elisabetta Ferretti and Francesco Dotta both share the senior authorship.
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Sebastiani, G., Po, A., Miele, E. et al. MicroRNA-124a is hyperexpressed in type 2 diabetic human pancreatic islets and negatively regulates insulin secretion. Acta Diabetol 52, 523–530 (2015). https://doi.org/10.1007/s00592-014-0675-y
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DOI: https://doi.org/10.1007/s00592-014-0675-y