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- Title
Systems Biology Identification of the AMPK-AKT-GSK3β Axis in Mediating Dysregulated Responses of Cystic Fibrosis Airway Cells to IDR-1018, an Immunomodulatory Peptide.
- Authors
Mayer, M. L.; Blohmke, C. J.; Fjell, C. D.; Turvey, S. E.; Hancock, R. E. W.
- Abstract
BACKGROUND: Innate defence regulator (IDR) peptides are synthetic derivatives of endogenous host defence peptides, and are currently under investigation as novel anti-infective agents. IDRs lack effective antimicrobial activity; rather, they enhance bacterial clearance by increasing leukocyte recruitment to infection sites while simultaneously suppressing harmful inflammation. As cystic fibrosis (CF) pathology arises from chronic bacterial infections which in turn drive chronic lung inflammation, we sought to investigate the ability of a lead IDR (IDR-1018) to modulate human bronchial epithelial (HBE) cell responses to pro-inflammatory stimuli. METHODS: Primary normal HBE cells, CF HBE cell lines (IB3-1, CuFi) or control cells (C38, NuLi, 16HBE) were stimulated for 24 hr with flagellin or heat-killed Pseudomonas aeruginosa, with or without 1 hr pretreatment with IDR-1018, and ELISA was used to measure IL-6 and IL-8 production. For microarray experiments, IB3-1 and C38 were stimulated with for 2 hr with flagellin, with or without 1 hr pretreatment with IDR-1018. RNA was harvested, converted to cDNA, and hybridized to Illumina HumanHT-12 chips. Systems biology and network analyses were carried out using our new systems biology platforms: InnateDB, Cerebral (Cytoscape) and MetaGEX. RESULTS: IDR-1018 was found to enhance IL-6 and IL-8 secretion induced by heat-killed P. aeruginosa or flagellin in normal cell lines but in contrast had an inhibitory effect on cytokine secretion in CF cell lines. Cytokine modulation by IDR-1018 was not observed in cells stimulated with PAK fliC, suggesting a flagellin-dependent effect. Systems biology and network analyses identified multiple dysregulated genes mediating the differences between normal and CF cells, including the AMPK-Akt-GSK-3β axis. Pretreating IB3-1 cells with metformin (AMPK activator) or inhibitors of Akt or GSK-3β had negligible or subtle inhibitory effects on flagellin-specific responses, but completely reversed the ability of IDR-1018 to inhibit airway cell cytokine secretion. CONCLUSION: Taken together, these data implicate the AMPK-Akt-GSK-3β axis in mediating dysfunctional responses of CF cells to immunomodulatory therapeutics, and validate the utility of systems biology in identifying signalling pathways which underlie abnormal pro-inflammatory cytokine secretion by CF cells.
- Subjects
SYSTEMS biology; CYSTIC fibrosis; IMMUNOREGULATION; PEPTIDES; INFLAMMATION; BIOLOGICAL transport; CYTOKINES; CELLS
- Publication
UBC Medical Journal, 2011, Vol 2, Issue 2, p54
- ISSN
1920-7425
- Publication type
Abstract