Virus-like infection induces human β cell dedifferentiation
Masaya Oshima, Klaus-Peter Knoch, Marc Diedisheim, Antje Petzold, Pierre Cattan, Marco Bugliani, Piero Marchetti, Pratik Choudhary, Guo-Cai Huang, Stefan R. Bornstein, Michele Solimena, Olivier Albagli-Curiel, Raphael Scharfmann
Masaya Oshima, Klaus-Peter Knoch, Marc Diedisheim, Antje Petzold, Pierre Cattan, Marco Bugliani, Piero Marchetti, Pratik Choudhary, Guo-Cai Huang, Stefan R. Bornstein, Michele Solimena, Olivier Albagli-Curiel, Raphael Scharfmann
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Research Article Inflammation Virology

Virus-like infection induces human β cell dedifferentiation

Abstract

Type 1 diabetes (T1D) is a chronic disease characterized by an autoimmune-mediated destruction of insulin-producing pancreatic β cells. Environmental factors such as viruses play an important role in the onset of T1D and interact with predisposing genes. Recent data suggest that viral infection of human islets leads to a decrease in insulin production rather than β cell death, suggesting loss of β cell identity. We undertook this study to examine whether viral infection could induce human β cell dedifferentiation. Using the functional human β cell line EndoC-βH1, we demonstrate that polyinosinic-polycytidylic acid (PolyI:C), a synthetic double-stranded RNA that mimics a byproduct of viral replication, induces a decrease in β cell–specific gene expression. In parallel with this loss, the expression of progenitor-like genes such as SOX9 was activated following PolyI:C treatment or enteroviral infection. SOX9 was induced by the NF-κB pathway and also in a paracrine non–cell-autonomous fashion through the secretion of IFN-α. Lastly, we identified SOX9 targets in human β cells as potentially new markers of dedifferentiation in T1D. These findings reveal that inflammatory signaling has clear implications in human β cell dedifferentiation.

Authors

Masaya Oshima, Klaus-Peter Knoch, Marc Diedisheim, Antje Petzold, Pierre Cattan, Marco Bugliani, Piero Marchetti, Pratik Choudhary, Guo-Cai Huang, Stefan R. Bornstein, Michele Solimena, Olivier Albagli-Curiel, Raphael Scharfmann

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Figure 2

PolyI:C induces the expression of SOX9, HES1, and MYC in human β cells.

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PolyI:C induces the expression of SOX9, HES1, and MYC in human β cells. ...
(A–C) EndoC-βH1 cells were either mock transfected (CTRL) or transfected with PolyI:C and analyzed 24 hours later (n = 3). (A and B) Heatmap from global transcriptomic analysis and RT-qPCR data represent induced genes. (C) Western blot analysis of SOX9 expression (n = 3; n1, n2, and n3 correspond to 3 independent experiments). (D) Primary human islet cells were dissociated and either mock transfected (CTRL) or transfected with PolyI:C and analyzed 24 hours later by immunocytochemistry. Nuclei are stained with Hoechst 33342 stain (blue), insulin is in red, and SOX9 is in green. Arrowheads point to insulin+ (INS+) cells that stain positive for SOX9 following PolyI:C treatment. Scale bars: 25 μm. The inset shows a higher-magnification image (×2.1 magnification; scale bar: 5 μm). Representative images of 4 independent experiments. (E–G) EndoC-βH1 cells were infected with enteroviruses at 5 × 104 TCID50 and harvested 24 hours later. (E) Western blot analysis of VP1 expression (n = 3). (F and G) qPCR analyses of SOX9, MYC, and HES1 expression (n = 3). (H) Human islets were infected with enteroviruses at 5 × 107 TCID50 and harvested 24 hours later. Western blot analyses of VP1 and SOX9 expression are shown (representative images of 3 independent experiments). Data from RT-qPCR, Western blots, and immunofluorescence represent the mean ± SD of 3 independent experiments. *P < 0.05, **P < 0.01, and ***P < 0.001 relative to control by Student’s t test. CVB5-F, coxsackievirus strain B5 Faulkner; EV-9 B, echovirus 9 Barty.