MDA5 and PTPN2, two candidate genes for type 1 diabetes, modify pancreatic -cell responses to the viral by-product double-stranded RNA

• Published in: Human molecular genetics 2010-01, Vol.19 (1), p.135-146
• Main Authors: Colli, Maikel L., Moore, Fabrice, Gurzov, Esteban N., Ortis, Fernanda, Eizirik, Decio L.
• Format: Article
• Language: English
• ISSN: 0964-6906; 1460-2083
• DOI: 10.1093/hmg/ddp474

-Cell destruction in type 1 diabetes (T1D) is at least in part consequence of a 'dialog' between -cells and immune system. This dialog may be affected by the individual's genetic background. We presently evaluated whether modulation of MDA5 and PTPN2, two candidate genes for T1D, affects -cell responses to double-stranded RNA (dsRNA), a by-product of viral replication. These genes were selected following comparison between known candidate genes for T1D and genes expressed in pancreatic -cells, as identified in previous array analysis. INS-1E cells and primary fluorescence-activated cell sorting-purified rat -cells were transfected with small interference RNAs (siRNAs) targeting MDA5 or PTPN2 and subsequently exposed to intracellular synthetic dsRNA (polyinosinic-polycitidilic acid-PIC). Real-time RT-PCR, western blot and viability assays were performed to characterize gene/protein expression and viability. PIC increased MDA5 and PTPN2 mRNA expression, which was inhibited by the specific siRNAs. PIC triggered apoptosis in INS-1E and primary -cells and this was augmented by PTPN2 knockdown (KD), although inhibition of MDA5 did not modify PIC-induced apoptosis. In contrast, MDA5 silencing decreased PIC-induced cytokine and chemokine expression, although inhibition of PTPN2 induced minor or no changes in these inflammatory mediators. These findings indicate that changes in MDA5 and PTPN2 expression modify -cell responses to dsRNA. MDA5 regulates inflammatory signals, whereas PTPN2 may function as a defence mechanism against pro-apoptotic signals generated by dsRNA. These two candidate genes for T1D may thus modulate -cell apoptosis and/or local release of inflammatory mediators in the course of a viral infection by acting, at least in part, at the pancreatic -cell level.