miR-17-92 and miR-106b-25 clusters regulate beta cell mitotic checkpoint and insulin secretion in mice
Adult beta cells in the pancreas are the sole source of insulin in the body. Beta cell loss or increased demand for insulin impose metabolic challenges because adult beta cells are generally quiescent and infrequently re-enter the cell division cycle. The aim of this study is to test the hypothesis that a family of proto-oncogene microRNAs that includes miR-17-92 and miR-106b-25 clusters regulates beta cell proliferation or function in the adult endocrine pancreas.
To elucidate the role of miR-17-92 and miR-106b-25 clusters in beta cells, we used a conditional miR-17-92/miR-106b-25 knockout mouse model. We employed metabolic assays in vivo and ex vivo, together with advanced microscopy of pancreatic sections, bioinformatics, mass spectrometry and next generation sequencing, to examine potential targets of miR-17-92/miR-106b-25, by which they might regulate beta cell proliferation and function.
We demonstrate that miR-17-92/miR-106b-25 regulate the adult beta cell mitotic checkpoint and that miR-17-92/miR-106b-25 deficiency results in reduction in beta cell mass in vivo. Furthermore, we reveal a critical role for miR-17-92/miR-106b-25 in glucose homeostasis and in controlling insulin secretion. We identify protein kinase A as a new relevant molecular pathway downstream of miR-17-92/miR-106b-25 in control of adult beta cell division and glucose homeostasis.
The study contributes to the understanding of proto-oncogene miRNAs in the normal, untransformed endocrine pancreas and illustrates new genetic means for regulation of beta cell mitosis and function by non-coding RNAs.
Sequencing data that support the findings of this study have been deposited in GEO with the accession code GSE126516.
KeywordsBeta cells Cell cycle Diabetes Glucose-stimulated insulin secretion GSIS microRNA PKA Protein kinase A
False discovery rate
Fluorescence resonance energy transfer
Glucose-stimulated insulin secretion
Microtubule affinity regulating kinase 2
Mouse embryonic fibroblast
Phosphorylated histone H3
Protein kinase A
Protein kinase cAMP-dependent type I regulatory subunit α
Quantitative real-time RT-PCR
Single molecule fluorescence in situ hybridisation
The authors would like to thank Y. Melamed and O. Higfa for veterinary services and husbandry (Weizmann Institute of Science). The authors thank A. Savidor and Y. Levin at the de Botton Institute for Protein Profiling, Weizmann Institute of Science, and O. Ben-Ami of the Crown Genomics Institute of the Nancy and Stephen Grand Israel National Center for Personalized Medicine, Weizmann Institute of Science, for MS and next generation sequencing, respectively. We thank O. Elhanani, M. Walker, S. Itzkovitz, H. Kaspi, R. Pasvolsky and E. Geron (Weizmann Institute of Science) for insightful comments on the manuscript.
ADM, SK and EH made substantial contributions to the conception or design of the work, the acquisition, analysis and interpretation of data and drafting the work for important intellectual content. YD provided substantial contribution to the conception or design of the work and drafting the work for important intellectual content and agrees to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved. EY, AK, AS, NM and DA contributed to acquisition, analysis or interpretation of data and drafting the work for important intellectual content. All authors gave final approval of the version to be published. EH is the guarantor of this work.
The work is funded by an ERC consolidator program (617351), Juvenile Diabetes Research Foundation (99-2007-71 and 47-2012-742), European Diabetes Research Programmes (EFSD)/D-Cure young Investigator award and EFSD-Lilly, Yeda-Sela, Yeda-CEO, Y. Leon Benoziyo Institute for Molecular Medicine, Kekst Family Institute for Medical Genetics, David and Fela Shapell Family Center for Genetic Disorders Research, Crown Human Genome Center, Nathan, Shirley, Philip and Charlene Vener New Scientist Fund, Julius and Ray Charlestein Foundation, Fraida Foundation, Wolfson Family Charitable Trust, Adelis Foundation, Merck (UK), Maria Halphen and Estates of Fannie Sherr, Lola Asseof, Lilly Fulop. Hornstein laboratory is supported by Dr. Sydney Brenner. EH is Head of Nella and Leon Benoziyo Center for Neurological Diseases and incumbent of Ira & Gail Mondry Professorial chair. The study sponsor was not involved in the design of the study, the collection, analysis and interpretation of data, writing the report or the decision to submit the report for publication.
Duality of interest
The authors declare that there is no duality of interest associated with this manuscript.
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