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Small-molecule studies identify CDK8 as a regulator of IL-10 in myeloid cells

Nature Chemical Biology volume 13, pages 1102–1108 (2017)Cite this article

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Abstract

Enhancing production of the anti-inflammatory cytokine interleukin-10 (IL-10) is a promising strategy to suppress pathogenic inflammation. To identify new mechanisms regulating IL-10 production, we conducted a phenotypic screen for small molecules that enhance IL-10 secretion from activated dendritic cells. Mechanism-of-action studies using a prioritized hit from the screen, BRD6989, identified the Mediator-associated kinase CDK8, and its paralog CDK19, as negative regulators of IL-10 production during innate immune activation. The ability of BRD6989 to upregulate IL-10 is recapitulated by multiple, structurally differentiated CDK8 and CDK19 inhibitors and requires an intact cyclin C–CDK8 complex. Using a highly parallel pathway reporter assay, we identified a role for enhanced AP-1 activity in IL-10 potentiation following CDK8 and CDK19 inhibition, an effect associated with reduced phosphorylation of a negative regulatory site on c-Jun. These findings identify a function for CDK8 and CDK19 in regulating innate immune activation and suggest that these kinases may warrant consideration as therapeutic targets for inflammatory disorders.

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Figure 1: Prioritization of BRD6989 from a phenotypic screen for small-molecule enhancers of IL-10 production.
Figure 2: CDK8 is a molecular target of BRD6989.
Figure 3: Pharmacological and genetic data support CDK8 as a negative regulator of IL-10 production.
Figure 4: Modulation of c-Jun–AP-1 links CDK8 inhibition to enhanced IL-10 production.

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Acknowledgements

This work was supported by funding from the National Institutes of Health Grants K08DK104021 (B.K.), R01CA190509 (P.S.), U01DK062432 and P30DK043351 (R.J.X.); Crohn's and Colitis Foundation of America Grant 500229 (R.J.X.); and The Leona M. and Harry B. Helmsley Charitable Trust Grant 500203 (S.L.S. and R.J.X.). S.L.S. is an Investigator of the Howard Hughes Medical Institute. We thank P. Baran (The Scripps Research Institute) for the generous gift of Δ16-cortistatin A.

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Author notes

  1. Liv Johannessen and Thomas B Sundberg: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts, USA

    Liv Johannessen, Baishan Jiang & Nathanael S Gray

  2. Department of Cancer Biology, Dana–Farber Cancer Institute, Boston, Massachusetts, USA

    Liv Johannessen, Anne Fassl, Baishan Jiang, Piotr Sicinski & Nathanael S Gray

  3. Center for the Development of Therapeutics, Broad Institute, Cambridge, Massachusetts, USA

    Thomas B Sundberg, James Berstler, Jose R Perez & Andrew J Phillips

  4. Infectious Disease and Microbiome Program, Broad Institute, Cambridge, Massachusetts, USA

    Daniel J O'Connell, Isabel J Latorre, Daniel B Graham & Ramnik J Xavier

  5. Center for Computational and Integrative Biology, Massachusetts General Hospital, Boston, Massachusetts, USA

    Raivo Kolde, Bernard Khor & Ramnik J Xavier

  6. Department of Chemistry, Yale University, New Haven, Connecticut, USA

    Katelyn J Billings

  7. Center for the Science of Therapeutics, Broad Institute, Cambridge, Massachusetts, USA

    Katelyn J Billings, Brinton Seashore-Ludlow, Stuart L Schreiber & Alykhan F Shamji

  8. Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA

    Anne Fassl & Piotr Sicinski

  9. Gastrointestinal Unit and Center for the Study of Inflammatory Bowel Disease, Massachusetts General Hospital, Boston, Massachusetts, USA

    Caitlin N Russell & Ramnik J Xavier

  10. Department of Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA

    Daniel B Graham

  11. Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA

    Stuart L Schreiber

  12. Howard Hughes Medical Institute, Cambridge, Massachusetts, USA

    Stuart L Schreiber

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Contributions

L.J. designed studies, conducted experiments, analyzed results and wrote the paper. T.B.S. devised the project, developed the screen, designed studies, conducted experiments, analyzed results and wrote the paper. D.J.O. designed studies, conducted experiments and analyzed results. R.K. analyzed gene expression and multiplex reporter data. J.B. designed studies, conducted experiments and analyzed results. K.J.B. synthesized BRD0330 analogs. B.K. designed studies, conducted experiments and analyzed results. B.S.-L. designed studies, conducted experiments and analyzed results. A.F. generated CcncΔ/Δ mice. C.N.R. conducted the screen. I.J.L. designed studies and analyzed results. B.J. synthesized CCT251921. D.B.G. designed studies, conducted experiments and analyzed results. J.R.P. designed studies, conducted experiments and analyzed results. P.S. generated CcncΔ/Δ mice. A.J.P. designed studies, conducted experiments and analyzed results. S.L.S. designed studies, analyzed results and wrote the paper. N.S.G. designed studies, analyzed results and wrote the paper. A.F.S. devised the project, developed the screen, synthesized CCT251921, designed studies, analyzed results and wrote the paper. R.J.X. devised the project, developed the screen, designed studies, analyzed results and wrote the paper.

Corresponding authors

Correspondence to Nathanael S Gray, Alykhan F Shamji or Ramnik J Xavier.

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Competing interests

L.J., T.B.S., B.K., J.R.P., N.S.G., A.F.S., and R.J.X. are co-inventors on a provisional patent application based on these results.

Supplementary information

Supplementary Text and Figures

Supplementary Results, Supplementary Tables 1–2 and Supplementary Figures 1–12. (PDF 4687 kb)

Supplementary Note

Chemical Synthesis of BRD10330 analogs. (PDF 375 kb)

Supplementary Data Set 1

BRD6989 Kinase profiling data (Life Technologies). (XLSX 22 kb)

Supplementary Data Set 2

BRD6989 Kinase profiling data (DiscoverX). (XLSX 59 kb)

Supplementary Data Set 3

Results from Gene Set Enrichment Analysis. (XLSX 14 kb)

Supplementary Data Set 4

Normalized TF-seq dataset. (XLSX 269 kb)

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Johannessen, L., Sundberg, T., O'Connell, D. et al. Small-molecule studies identify CDK8 as a regulator of IL-10 in myeloid cells. Nat Chem Biol 13, 1102–1108 (2017). https://doi.org/10.1038/nchembio.2458

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