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Negative regulation of TLR4 via targeting of the proinflammatory tumor suppressor PDCD4 by the microRNA miR-21

Nature Immunology volume 11, pages 141–147 (2010)Cite this article

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Abstract

The tumor suppressor PDCD4 is a proinflammatory protein that promotes activation of the transcription factor NF-κB and suppresses interleukin 10 (IL-10). Here we found that mice deficient in PDCD4 were protected from lipopolysaccharide (LPS)-induced death. The induction of NF-κB and IL-6 by LPS required PDCD4, whereas LPS enhanced IL-10 induction in cells lacking PDCD4. Treatment of human peripheral blood mononuclear cells with LPS resulted in lower PDCD4 expression, which was due to induction of the microRNA miR-21 via the adaptor MyD88 and NF-κB. Transfection of cells with a miR-21 precursor blocked NF-κB activity and promoted IL-10 production in response to LPS, whereas transfection with antisense oligonucleotides to miR-21 or targeted protection of the miR-21 site in Pdcd4 mRNA had the opposite effect. Thus, miR-21 regulates PDCD4 expression after LPS stimulation.

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Figure 1: PDCD4-deficient mice are protected from the lethality of LPS.
Figure 2: PDCD4 protein expression is regulated by LPS.
Figure 3: Regulation of TLR signaling by PDCD4.
Figure 4: Induction of miR-21 by LPS treatment in macrophages.
Figure 5: Regulation of PDCD4 expression in LPS signaling.
Figure 6: Regulation of PDCD4 function by miR-21 in TLR signaling (a) ELISA of mouse IL-10 in RAW264.7 cells transfected with various doses (horizontal axes) of pro-miR-21 (left), anti-miR-21 (middle) or morpho-21 (right) and then stimulated for 24 h with LPS (100 ng/ml).

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  • 06 December 2009

    In the version of this article initially published online, several author names and affiliations are incorrect. The correct names and affiliations are as follows: Frederick J Sheedy1, Eva Palsson-McDermott1, Elizabeth J Hennessy1, Cara Martin2,3, John J O'Leary2,3, Qingguo Ruan4, Derek S Johnson4, Youhai Chen4 & Luke A J O'Neill1 1School of Biochemistry & Immunology, Trinity College, Dublin, Ireland. 2Department of Pathology, Coombe Women's Hospital, Dublin, Ireland. 3Department of Histopathology, School of Medicine, Trinity College, Dublin, Ireland. 4Department of Pathology and Laboratory Medicine, School of Medicine, University of Pennsylvania, Philadelphia, USA. The errors have been corrected for the print, PDF and HTML versions of this article.

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Acknowledgements

We thank D. Golenbock (University of Massachusetts) for wild-type, MyD88-deficient and TRIF-deficient BMDMs immortalized by retrovirus; R. Hay (University of St. Andrews) for p65-deficient and matched wild-type control mouse embryonic fibroblasts and for HA-tagged ubiquitin; R. Hofmeister (Universitaet Regensburg) for 5× NF-κB reporter luciferase plasmid; A.M. Cheng (Ambion) for the pMIR-REPORT miR-21 reporter luciferase plasmid; and A. Bowie (Trinity College, Dublin) for the Il10 promoter–luciferase plasmid. Supported by Science Foundation Ireland and the Irish Research Council for Science, Engineering and Technology (RS/2005/190).

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Authors and Affiliations

  1. School of Biochemistry & Immunology, Trinity College, Dublin, Ireland

    Frederick J Sheedy, Eva Palsson-McDermott, Elizabeth J Hennessy & Luke A J O'Neill

  2. Department of Pathology, Coombe Women's Hospital, Dublin, Ireland

    Cara Martin & John J O'Leary

  3. Department of Histopathology, School of Medicine, Trinity College, Dublin, Ireland

    Cara Martin & John J O'Leary

  4. Department of Pathology and Laboratory Medicine, School of Medicine, University of Pennsylvania, Philadelphia, USA

    Qingguo Ruan, Derek S Johnson & Youhai Chen

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Contributions

F.J.S. did the functional experiments on PDCD4 and miR-21 and cowrote the manuscript; E.P.-M. did experiments on PDCD4 degradation by the proteasome and experiments on signals in PDCD4-deficient cells; E.J.H. helped with experiments on miR-21; C.M. and J.J.O. provided advice on miRNA profiling experiments; Q.R., D.S.J. and Y.C. did the experiments on the lethality of LPS in PDCD4–deficient mice and supplied BMDMs from the mice; and L.A.J.O. directed the work and cowrote the manuscript.

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Correspondence to Luke A J O'Neill.

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Sheedy, F., Palsson-McDermott, E., Hennessy, E. et al. Negative regulation of TLR4 via targeting of the proinflammatory tumor suppressor PDCD4 by the microRNA miR-21. Nat Immunol 11, 141–147 (2010). https://doi.org/10.1038/ni.1828

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