title: An Activating STAT3 Mutation Causes Neonatal Diabetes through Premature Induction of Pancreatic Differentiation.
abstract: Activating germline mutations in STAT3 were recently identified as a cause of neonatal diabetes mellitus associated with beta-cell autoimmunity. We have investigated the effect of an activating mutation, STAT3K392R, on pancreatic development using induced pluripotent stem cells (iPSCs) derived from a patient with neonatal diabetes and pancreatic hypoplasia. Early pancreatic endoderm differentiated similarly from STAT3K392Rand healthy-control cells, but in later stages, NEUROG3 expression was upregulated prematurely in STAT3K392Rcells together with insulin (INS) and glucagon (GCG). RNA sequencing (RNA-seq) showed robust NEUROG3 downstream targets upregulation. STAT3 mutation correction with CRISPR/Cas9 reversed completely the disease phenotype. STAT3K392R-activating properties were not explained fully by altered DNA-binding affinity or increased phosphorylation. Instead, reporter assays demonstrated NEUROG3 promoter activation by STAT3 in pancreatic cells. Furthermore, proteomic and immunocytochemical analyses revealed increased nuclear translocation of STAT3K392R. Collectively, our results demonstrate that the STAT3K392Rmutation causes premature endocrine differentiation through direct induction of NEUROG3 expression.
authors.affiliation: Research Programs Unit, Molecular Neurology and Biomedicum Stem Cell Centre, Faculty of Medicine, University of Helsinki, 00014 Helsinki, Finland. Electronic address: jonna.saarimaki@helsinki.fi.
authors.forename: Jonna
authors.initials: J
authors.lastname: Saarimäki-Vire
authors.affiliation: Research Programs Unit, Molecular Neurology and Biomedicum Stem Cell Centre, Faculty of Medicine, University of Helsinki, 00014 Helsinki, Finland.
authors.forename: Diego
authors.initials: D
authors.lastname: Balboa
authors.affiliation: Institute of Biomedical and Clinical Science, University of Exeter Medical School, Exeter EX2 5DW, UK.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Russell
authors.affiliation: Faculty of Medicine and Life Sciences, University of Tampere, and Tampere University Hospital, 33014 Tampere, Finland.
authors.forename: Juha
authors.initials: J
authors.lastname: Saarikettu
authors.affiliation: Molecular Systems Biology Research Group, Institute of Biotechnology, University of Helsinki, 00014 Helsinki, Finland.
authors.forename: Matias
authors.initials: M
authors.lastname: Kinnunen
authors.affiliation: Molecular Systems Biology Research Group, Institute of Biotechnology, University of Helsinki, 00014 Helsinki, Finland.
authors.forename: Salla
authors.initials: S
authors.lastname: Keskitalo
authors.affiliation: Institute of Biomedical and Clinical Science, University of Exeter Medical School, Exeter EX2 5DW, UK.
authors.forename: Amrinder
authors.initials: A
authors.lastname: Malhi
authors.affiliation: Division of Medical Genetics, Department of Medicine, Department of Genome Sciences, Institute for Stem Cell and Regenerative Medicine, University of Washington School of Medicine, Seattle, WA 98109, USA.
authors.forename: Cristina
authors.initials: C
authors.lastname: Valensisi
authors.affiliation: Division of Medical Genetics, Department of Medicine, Department of Genome Sciences, Institute for Stem Cell and Regenerative Medicine, University of Washington School of Medicine, Seattle, WA 98109, USA.
authors.forename: Colin
authors.initials: C
authors.lastname: Andrus
authors.affiliation: Research Programs Unit, Molecular Neurology and Biomedicum Stem Cell Centre, Faculty of Medicine, University of Helsinki, 00014 Helsinki, Finland.
authors.forename: Solja
authors.initials: S
authors.lastname: Eurola
authors.affiliation: Research Programs Unit, Molecular Neurology and Biomedicum Stem Cell Centre, Faculty of Medicine, University of Helsinki, 00014 Helsinki, Finland.
authors.forename: Heli
authors.initials: H
authors.lastname: Grym
authors.affiliation: Research Programs Unit, Molecular Neurology and Biomedicum Stem Cell Centre, Faculty of Medicine, University of Helsinki, 00014 Helsinki, Finland.
authors.forename: Jarkko
authors.initials: J
authors.lastname: Ustinov
authors.affiliation: Research Programs Unit, Molecular Neurology and Biomedicum Stem Cell Centre, Faculty of Medicine, University of Helsinki, 00014 Helsinki, Finland; Clinical Genetics, HUSLAB, Helsinki University Central Hospital, Haartmaninkatu 4, 00290 Helsinki, Finland.
authors.forename: Kirmo
authors.initials: K
authors.lastname: Wartiovaara
authors.affiliation: Division of Medical Genetics, Department of Medicine, Department of Genome Sciences, Institute for Stem Cell and Regenerative Medicine, University of Washington School of Medicine, Seattle, WA 98109, USA.
authors.forename: R David
authors.initials: RD
authors.lastname: Hawkins
authors.affiliation: Faculty of Medicine and Life Sciences, University of Tampere, and Tampere University Hospital, 33014 Tampere, Finland.
authors.forename: Olli
authors.initials: O
authors.lastname: Silvennoinen
authors.affiliation: Molecular Systems Biology Research Group, Institute of Biotechnology, University of Helsinki, 00014 Helsinki, Finland.
authors.forename: Markku
authors.initials: M
authors.lastname: Varjosalo
authors.affiliation: Institute of Biomedical and Clinical Science, University of Exeter Medical School, Exeter EX2 5DW, UK.
authors.forename: Noel G
authors.initials: NG
authors.lastname: Morgan
authors.affiliation: Research Programs Unit, Molecular Neurology and Biomedicum Stem Cell Centre, Faculty of Medicine, University of Helsinki, 00014 Helsinki, Finland; Children's Hospital, University of Helsinki and Helsinki University Hospital, 00290 Helsinki, Finland. Electronic address: timo.otonkoski@helsinki.fi.
authors.forename: Timo
authors.initials: T
authors.lastname: Otonkoski
mesh_terms: D015551:Autoimmunity / Q000235:genetics; D051792:Basic Helix-Loop-Helix Proteins / Q000096:biosynthesis* / Q000235:genetics; D064113:CRISPR-Cas Systems; D002454:Cell Differentiation / Q000235:genetics*; D002460:Cell Line; D003920:Diabetes Mellitus / Q000209:etiology / Q000235:genetics* / Q000473:pathology; D018507:Gene Expression Regulation, Developmental; D005934:Glucagon / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism / Q000473:pathology; D007328:Insulin / Q000235:genetics / Q000378:metabolism; D050417:Insulin-Secreting Cells / Q000378:metabolism / Q000473:pathology; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000096:biosynthesis* / Q000235:genetics; D011401:Promoter Regions, Genetic; D050796:STAT3 Transcription Factor / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D005934:Glucagon; D007328:Insulin; D009419:Nerve Tissue Proteins; D050796:STAT3 Transcription Factor; C498695:NEUROG3 protein, human; C494086:STAT3 protein, human
keywords: CRISPR; NEUROG3; STAT3; beta cell; endocrine cells; genome editing; iPSC; monogenic diabetes; pancreatic differentiation; stem cells