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, STAT3, 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 STAT3 and healthy-control cells, but in later stages, NEUROG3 expression was upregulated prematurely in STAT3 cells 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. STAT3-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 STAT3. Collectively, our results demonstrate that the STAT3 mutation causes premature endocrine differentiation through direct induction of NEUROG3 expression.

Pancreas patch-seq links physiologic dysfunction in diabetes to single-cell transcriptomic phenotypes

“…Our work suggests that ubiquitination and proteosomal degradation pathways in b-cells, including a pathway stimulating COP1 auto-degradation, are associated with dysfunction in T2D (Fig 4; Supp Fig 10). Thus, in addition to transcript-based mechanisms, our results suggest that post-translational mechanisms involving factors like COP1, ETV1 and STAT3 may govern b-cell dysfunction or responses in T2D (O'Shea and Plenge, 2012;Saarimäki-Vire et al, 2017;Suriben et al, 2015).…”

“…We tested genes previously reported to show ETVdependent expression in mouse islets (Suriben et al, 2015), and found several human orthologs in our bcell single-cell correlations, including STAT3, MAFA, SLC30A8. Analysis of these ETV-dependent genes showed an enrichment of (i) pathways that we found to be important for b-cell exocytosis in our correlation analysis, and (ii) genes related to immune cytokine signaling, like STAT3 which encodes a critical cytokine signaling factor, that also has postulated roles in insulin secretion, b-cell regeneration and neonatal diabetes (O'Shea and Plenge, 2012;Saarimäki-Vire et al, 2017). Finally, we analyzed fulllength transcriptomes to investigate ETV1 splicing in b-cells and detected no signs of differential splicing between ND and T2D (Supp Fig 10).…”

Exenatide induces frataxin expression and improves mitochondrial function in Friedreich ataxia

“…Recent developments in iPSC differentiation make it possible to generate functional sensory neurons using a 1-step protocol (46). We implemented this technology to differentiate sensory neurons from FRDA patients' iPSCs FA135 and HEL135 and also used 2 previously characterized control iPSCs, HEL115.6 and HEL46.11 (47,48). At the end of the 8-day differentiation, most cells expressed sensory neuron-specific markers Brn3a, peripherin, and ISL1 and neuron-specific cytoskeletal marker β-tubulin III, showing sensory neuron lineage; expression of these markers was confirmed at the mRNA level ( Figure 5A and Supplemental Figure 7, A-E).…”

Postoperative resveratrol administration improves prognosis of rat orthotopic glioblastomas

“…Cells apoptosis in the tumor tissues was tested by terminal deoxynucleotide transferase(TdT)–mediated dUTP-biotin nick-end labeling (TUNEL) according to manufacturer’s instructions (Roche Diagnostics GmbH, Mannheim, Germany). STAT3 signaling as the critical survival factor of glioblastoma cells and the main molecular target of resveratrol [22, 23], the status of STAT3 signaling in the tumor tissues with and without LP resveratrol treatment was detected by Western blotting and IHC staining using rabbit anti-STAT3 antibody (1:1000 for Western blotting and 1:300 for IHC staining; Santa Cruz, CA, USA, Santa Cruz Biotech. Inc.), mouse anti-phosphorylated STAT3 antibody (1:600 for Western blotting and 1:300 for IHC staining; Chicago, IL, USA, ProteinTech Inc.) and rabbit anti-PIAS3 antibody (1:1000 for Western blotting and 1:300 for IHC staining; Santa Cruz, CA, USA, Santa Cruz Biotech.…”