Date: 05 Nov 2003
Global profiling of double stranded RNA- and IFN-γ-induced genes in rat pancreatic beta cells
Viral infections and local production of IFN-γ might contribute to beta-cell dysfunction/death in Type 1 Diabetes. Double stranded RNA (dsRNA) accumulates in the cytosol of viral-infected cells, and exposure of purified rat beta cells to dsRNA (tested in the form of polyinosinic-polycytidylic acid, PIC) in combination with IFN-γ results in beta-cell dysfunction and apoptosis. To elucidate the molecular mechanisms involved in PIC + IFN-γ-effects, we determined the global profile of genes modified by these agents in primary rat beta cells.
FACS-purified rat beta cells were cultured for 6 or 24 h in control condition or with IFN-γ, PIC or a combination of both agents. The gene expression profile was analysed in duplicate by high-density oligonucleotide arrays representing 5000 full-length genes and 3000 EST’s. Changes of greater than or equal to 2.5-fold were considered as relevant.
Following a 6- or 24-h treatment with IFN-γ, PIC or IFN-γ and PIC, we observed changes in the expression of 51 to 189 genes. IFN-γ modified the expression of MHC-related genes, and also of genes involved in beta-cell metabolism, protein processing, cytokines and signal transduction. PIC affected preferentially the expression of genes related to cell adhesion, cytokines and dsRNA signal transduction, transcription factors and MHC. PIC and/or IFN-γ up-regulated the expression of several chemokines and cytokines that could contribute to mononuclear cell homing and activation during viral infection, while IFN-γ induced a positive feedback on its own signal transduction. PIC + IFN-γ inhibited insulin and GLUT-2 expression without modifying pdx-1 mRNA expression.
This study provides the first comprehensive characterization of the molecular responses of primary beta cells to dsRNA + IFN-γ, two agents that are probably present in the beta cell milieu during the course of virally-induced insulitis and Type 1 Diabetes. Based on these findings, we propose an integrated model for the molecular mechanisms involved in dsRNA + IFN-γ induced beta-cell dysfunction and death.
J. Rasschaert and D. Liu contributed equally to the article
- Global profiling of double stranded RNA- and IFN-γ-induced genes in rat pancreatic beta cells
Volume 46, Issue 12 , pp 1641-1657
- Cover Date
- Print ISSN
- Online ISSN
- Additional Links
- Double stranded RNA
- microarray analysis
- pancreatic beta cells
- nitric oxide
- diabetes mellitus
- Industry Sectors
- Author Affiliations
- 1. Laboratory of Experimental Medicine, Université Libre de Bruxelles, Route de Lennik, 808, CP 618, 1070, Brussels, Belgium
- 2. Molecular Diagnostic Laboratory, Department of Clinical Biochemistry, Aarhus University, Denmark