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 2

ZNF148 is not required for β cells specification.

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ZNF148 is not required for β cells specification. (A) Schematic view of...
(A) Schematic view of ZNF148 mRNA variants, and sgRNA sequence targeting exon 7. Edited sequences from ZNF148-KO clone A and clone B lines are displayed below, with indels in red. (B) RNA-Seq measurement of ZNF148 (transcripts per million) in unedited (gold) and edited (cyan) eBC (clone A). (C) Immunofluorescence staining of ZNF148 (red) and INS (green) in immature β-like clusters (day 20). DAPI staining depicts nuclei (blue). Scale bar: 50 μm. (D) Representative image of flow cytometry quantification of GFP+ live cells in control β-like clusters and ZNF148-KO clusters (clone A) (left panels), and quantitative analysis indicating percentage of GFP+ cells in control and ZNF148-KO β-like clusters (clone A and B) (right panel). n = 11–37. (E) INS mRNA expression levels in control and ZNF148-KO β-like clusters (clone A and B) as measured by qPCR. n = 3–6. (F) Representative image of flow cytometry quantification of C-PEP+, NKX6.1+, PDX1+, C-PEP+/NKX6.1+, and C-PEP+/PDX1+ double-positive cells in control and ZNF148-KO β-like clusters (clone A). (G) Quantitative flow cytometry analysis indicating percentage of C-PEP+, NKX6.1+, PDX1+, and C-PEP+/NKX6.1+ double-positive cells in control and ZNF148-KO β-like clusters (clone A). n = 4. (H) Bright-field and fluorescence representative images of control and ZNF148-KO (clone A) β-like clusters (day 20). Scale bar: 200 μm.