An Activating STAT3 Mutation Causes Neonatal Diabetes through Premature Induction of Pancreatic Differentiation

• DOI: 10.1016/j.celrep.2017.03.055
• Publication Date: 2017-04-11T18:08:40Z
• Authors: Jonna Saarimäki-Vire; Diego Balboa; Mark A. Russell; Juha Saarikettu; Matias Kinnunen; Salla Keskitalo; Amrinder Malhi; Cristina Valensisi; Colin Andrus; Solja Eurola; Heli Grym; Jarkko Ustinov; Kirmo Wartiovaara; R. David Hawkins; Olli Silvennoinen; Markku Varjosalo; Noel G. Morgan; Timo Otonkoski
• Keywords: Autoimmunity; STAT3; NEUROG3; Insulin-Secreting Cells; Basic Helix-Loop-Helix Transcription Factors; Insulin; Developmental; Biology (General); Promoter Regions, Genetic; 0303 health sciences; iPSC; Gene Expression Regulation, Developmental; Cell Differentiation; Wessex Classification Subject Headings::Oncology. Pathology.::Genetics; ENDOCRINE PANCREAS; 3. Good health; CRISPR; monogenic diabetes; PLURIPOTENT STEM-CELLS; NEUROGENIN3; EXPRESSION; STAT3 Transcription Factor; QH301-705.5; Induced Pluripotent Stem Cells; 610; Nerve Tissue Proteins; Cell Line; Promoter Regions; 03 medical and health sciences; Genetic; stem cells; REVEALS; Diabetes Mellitus; genome editing; Humans; Biochemistry, cell and molecular biology; BETA-LIKE CELLS; pancreatic differentiation; IN-VITRO; endocrine cells; Glucagon; beta cell; Biomedicine; Gene Expression Regulation; PROGENITOR CELLS; Mutation; CRISPR-Cas Systems; KNOCKOUT MICE; GENERATION

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 an activating mutation, STAT3K392R, on pancreatic development using induced pluripotent stem cells (iPSCs) derived from patient and hypoplasia. Early endoderm differentiated similarly STAT3K392R healthy-control cells, but later stages, NEUROG3 expression was upregulated prematurely together insulin (INS) glucagon (GCG). RNA sequencing (RNA-seq) showed robust downstream targets upregulation. mutation correction CRISPR/Cas9 reversed completely disease phenotype. STAT3K392R-activating properties not explained fully by altered DNA-binding affinity or increased phosphorylation. Instead, reporter assays demonstrated promoter activation cells. Furthermore, proteomic immunocytochemical analyses revealed nuclear translocation STAT3K392R. Collectively, our results demonstrate that causes premature endocrine differentiation through direct induction expression.