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A combinatorial F box protein directed pathway controls TRAF adaptor stability to regulate inflammation

Nature Immunology volume 14, pages 470–479 (2013)Cite this article

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Abstract

Uncontrolled activation of tumor necrosis factor receptor–associated factor (TRAF) proteins may result in profound tissue injury by linking surface signals to cytokine release. Here we show that a ubiquitin E3 ligase component, Fbxo3, potently stimulates cytokine secretion from human inflammatory cells by destabilizing a sentinel TRAF inhibitor, Fbxl2. Fbxo3 and TRAF protein in circulation positively correlated with cytokine responses in subjects with sepsis, and we identified a polymorphism in human Fbxo3, with one variant being hypofunctional. A small-molecule inhibitor targeting Fbxo3 was sufficient to lessen severity of cytokine-driven inflammation in several mouse disease models. These studies identified a pathway of innate immunity that may be useful to detect subjects with altered immune responses during critical illness or provide a basis for therapeutic intervention targeting TRAF protein abundance.

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Figure 1: Fbxl2 interacts with and targets TRAFs for polyubiquitination.
Figure 2: Fbxl2 is phosphorylated and targeted by the SCF E3 ligase subunit Fbxo3.
Figure 3: Fbxo3 contains a naturally occurring polymorphism at Val221.
Figure 4: Fbxo3(V221I) is a loss-of-function polymorphism of Fbxo3 in vitro.
Figure 5: Fbxo3 knockdown ameliorates Pseudomonas-induced lung injury.
Figure 6: F box and TRAF proteins in subjects with sepsis.
Figure 7: Fbxo3 inhibitor BC-1215 lowers bacterial-induced inflammation.
Figure 8: Molecular regulation of proinflammatory cytokines mediated by F box proteins.

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Acknowledgements

We thank A. Lagneaux and S. Barge for assistance with studies. This material is based upon work supported, in part, by the US Department of Veterans Affairs, Veterans Health Administration, Office of Research and Development, Biomedical Laboratory Research and Development. This work was supported by a Merit Review Award from the US Department of Veterans Affairs and National Institutes of Health R01 grants HL096376, HL097376, HL081784, HL068135 and HL098174 (to R.K.M.), HL116472 (to B.B.C.), HL01916 (to Y.Z.) and P50HL084948 (F.C.S.), American Heart Association awards 12SDG9050005 (to J.Z.) and 12SDG12040330 (to C.Z.), and American Heart Association Grant-in-Aid 12GRNT11820019 (B.J.M.). The contents presented do not represent the views of the Department of Veterans Affairs or the United States Government.

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Authors and Affiliations

  1. Department of Medicine, Acute Lung Injury Center of Excellence, University of Pittsburgh, Pittsburgh, Pennsylvania, USA

    Bill B Chen, Tiffany A Coon, Jennifer R Glasser, Bryan J McVerry, Jing Zhao, Yutong Zhao, Chunbin Zou, Bryon Ellis, Frank C Sciurba, Yingze Zhang & Rama K Mallampalli

  2. Department of Cell Biology and Physiology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA

    Rama K Mallampalli

  3. Medical Specialty Service Line, Veterans Affairs Pittsburgh Healthcare System, Pittsburgh, Pennsylvania, USA

    Rama K Mallampalli

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Contributions

R.K.M. and B.B.C. jointly oversaw the studies. B.B.C. designed the study, performed experiments, analyzed the data and wrote the manuscript; T.A.C., J.R.G., J.Z, Y.Z. and C.Z. performed experiments and assisted with animal experiments; B.J.M., B.E., F.C.S. and Y.Z. assisted with human studies. R.K.M. revised the manuscript and directed the study.

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Correspondence to Bill B Chen or Rama K Mallampalli.

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Competing interests

A provisional patent application (US 61/657,423) covering Fbxo3 inhibitors and additional modifications was filed jointly through the United States Department of Veterans Affairs and the University of Pittsburgh.

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Chen, B., Coon, T., Glasser, J. et al. A combinatorial F box protein directed pathway controls TRAF adaptor stability to regulate inflammation. Nat Immunol 14, 470–479 (2013). https://doi.org/10.1038/ni.2565

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