Loss of ZNF148 enhances insulin secretion in human pancreatic β cells
Eleonora de Klerk, Yini Xiao, Christopher H. Emfinger, Mark P. Keller, David I. Berrios, Valentina Loconte, Axel A. Ekman, Kate L. White, Rebecca L. Cardone, Richard G. Kibbey, Alan D. Attie, Matthias Hebrok
Eleonora de Klerk, Yini Xiao, Christopher H. Emfinger, Mark P. Keller, David I. Berrios, Valentina Loconte, Axel A. Ekman, Kate L. White, Rebecca L. Cardone, Richard G. Kibbey, Alan D. Attie, Matthias Hebrok
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Research Article Metabolism Stem cells

Loss of ZNF148 enhances insulin secretion in human pancreatic β cells

Abstract

Insulin secretion from pancreatic β cells is essential to the maintenance of glucose homeostasis. Defects in this process result in diabetes. Identifying genetic regulators that impair insulin secretion is crucial for the identification of novel therapeutic targets. Here, we show that reduction of ZNF148 in human islets, and its deletion in stem cell–derived β cells (SC–β cells), enhances insulin secretion. Transcriptomics of ZNF148-deficient SC–β cells identifies increased expression of annexin and S100 genes whose proteins form tetrameric complexes involved in regulation of insulin vesicle trafficking and exocytosis. ZNF148 in SC–β cells prevents translocation of annexin A2 from the nucleus to its functional place at the cell membrane via direct repression of S100A16 expression. These findings point to ZNF148 as a regulator of annexin-S100 complexes in human β cells and suggest that suppression of ZNF148 may provide a novel therapeutic strategy to enhance insulin secretion.

Authors

Eleonora de Klerk, Yini Xiao, Christopher H. Emfinger, Mark P. Keller, David I. Berrios, Valentina Loconte, Axel A. Ekman, Kate L. White, Rebecca L. Cardone, Richard G. Kibbey, Alan D. Attie, Matthias Hebrok

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Figure 1

Characterization of ZNF148 expression during SC–β cell differentiation.

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Characterization of ZNF148 expression during SC–β cell differentiation. ...
(A) Schematic representation of the differentiation process from hESC (day 0) to eBC (day 27). (B) qRT-PCR time course expression analysis of ZNF148 and INS from undifferentiated hESCs (day 0) to immature β-like clusters (day 20). Data for ZNF148 and INS are presented as mean fold change relative to day 0 and to day 12, respectively; n = 3 independent and n = 2 technical samples. (C) qRT-PCR for ZNF148 and INS mRNA in pancreatic progenitors (day 12) and immature β-like clusters (day 20). Data presented as mean fold change relative to day 12; n = 3 independent and n = 2 technical samples. Data are presented as mean ± SEM. ***P < 0.001 determined by 2-tailed Student’s t test. (D) FPKM values from RNA-Seq data for ZNF148 mRNA in immature β-like clusters (day 20), eBC (day 27), and endogenous β cells isolated from human islets by fluorescence-activated cell sorting (FACS). Data presented as mean ± SD; n = 2 independent samples. (E) Immunofluorescent staining of ZNF148 (red) in eBC. DAPI depicts nuclei (blue). Scale bar: 50 μm. (F and G) t-Distributed stochastic neighbor embedding algorithm (t-SNE) analysis from scRNA-Seq of immature β-like clusters (F) and eBC (G). Cells expressing INS, GCG, SST, and ZNF148 are shown in separate plots, with expression levels shown as heatmap of log2 expression. Expression levels in β-like clusters: INS log2 expression (min 3.3, max 12.2), GCG log2 expression (min 0, max 12.4), SST log2 expression (min 1, max 13.6), ZNF148 log2 expression (min 1, max 3). Expression levels in eBC: INS log2 expression (min 5.3, max 12.4), GCG log2 expression (min 0, max 12.4), SST log2 expression (min 2.3, max 13.3), ZNF148 log2 expression (min 0, max 2.8). NS, not significant.