Abstract
Neuroinflammation associated with degenerative central nervous system disease and injury frequently results in oligodendrocyte death. While promoting oligodendrocyte viability is a major therapeutic goal, little is known about protective signaling strategies. We report that in highly purified rat oligodendrocytes, interferon gamma (IFNγ) activates a signaling pathway that protects these cells from tumor necrosis factor alpha (TNFα)-induced cytotoxicity. IFNγ protection requires Jak (Janus kinase) activation, components of the integrated stress response and NF-κB activation. Although NF-κB activation also occurred transiently in the absence of IFNγ and presence of TNFα, this activation was not sufficient to prevent induction of the TNFα-responsive cell death pathway. Genetic inhibition of NF-κB translocation to the nucleus abrogated IFNγ-mediated protection and did not change the cell death induced by TNFα, suggesting that NF-κB activation via IFNγ induces a different set of responses than activation of NF-κB via TNFα. A promising candidate is the NF-κB target cFLIP (cellular FLICE (FADD-like IL-1β-converting enzyme)-inhibitory protein), which is protease-deficient caspase homolog that inhibits caspase-3 activation. We show that IFNγ-mediated protection led to upregulation of cFLIP. Overexpression of cFLIP was sufficient for oligodendrocyte protection from TNFα and short hairpin RNA knockdown of cFLIP-abrogated IFNγ -mediated protection. To determine the relevance of our in vitro finding to the more complex in vivo situation, we determined the impact on oligodendrocyte death of regional cFLIP loss of function in a murine model of neuroinflammation. Our data show that downregulation of cFLIP during inflammation leads to death of oligodendrocytes and decrease of myelin in vivo. Taken together, we show that IFNγ-mediated induction of cFLIP expression provides a new mechanism by which this cytokine can protect oligodendrocytes from TNFα-induced cell death.
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Abbreviations
- IFN-γ:
-
interferon gamma
- TNFα:
-
tumor necrosis factor alpha
- Jak:
-
Janus kinase
- ISR:
-
integrated stress response
- cFLIP:
-
cellular FLICE (FADD-like IL-1β-converting enzyme)-inhibitory protein
- CNS:
-
central nervous system
- O-2A/OPC:
-
oligodendrocyte progenitor cells
- EAE:
-
experimental autoimmune encephalomyelitis
- PKR:
-
double-stranded RNA-dependent protein kinase R
- GalC:
-
galactocerebroside
- CM:
-
conditioned medium
- STATs:
-
signal transducer and activators of transcription proteins
- IRF-1:
-
IFN response factor 1
- SOCS:
-
suppressor of cytokine signaling
- UPR:
-
unfolded protein response
- eIF2α:
-
eukaryotic translation initiation factor-α
- PERK:
-
PKR-like endoplasmic reticulum kinase
- IκB-SR:
-
IκB-super repressor.
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Acknowledgements
This work was supported by a New York State Department of Health Award (grant no. CO23691–MMP). DCT was supported in part by a Kirchstein NRSA fellowship (NINDS; T32NS051152) and a postdoctoral fellowship from the NMSS (FG1812A1). The authors are grateful for insight and comments from Drs. Fred Strathmann, Ibro Ambeskovic, Brett Stevens and Christoph Proschel, and thank Jonathan D Cherry, Brendan Carlin and Addie Bardin for their expert technical assistance.
Author Contributions
DCT designed and conducted experiments, formulated the underlying hypothesis and wrote the manuscript draft. AC designed and conducted experiments, and contributed to the writing of the manuscript. MM-P conducted experiments, provided resources, contributed to the design, analysis and interpretation of data. KMO’B provided the IL-1βXAT mice, and contributed to the design and interpretation of the in vivo data. MN provided resources, and was involved in the writing and editing of the manuscript, and providing critically intellectual content.
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Tanner, D., Campbell, A., O'Banion, K. et al. cFLIP is critical for oligodendrocyte protection from inflammation. Cell Death Differ 22, 1489–1501 (2015). https://doi.org/10.1038/cdd.2014.237
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DOI: https://doi.org/10.1038/cdd.2014.237
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