Abstract

Interaction of pathogens with cells of the immune system results in activation of inflammatory gene expression. This response, although vital for immune defence, is frequently deleterious to the host due to the exaggerated production of inflammatory proteins. The scope of inflammatory responses reflects the activation state of signalling proteins upstream of inflammatory genes as well as signal-induced assembly of nuclear chromatin complexes that support mRNA expression1,2,3,4. Recognition of post-translationally modified histones by nuclear proteins that initiate mRNA transcription and support mRNA elongation is a critical step in the regulation of gene expression5,6,7,8,9,10. Here we present a novel pharmacological approach that targets inflammatory gene expression by interfering with the recognition of acetylated histones by the bromodomain and extra terminal domain (BET) family of proteins. We describe a synthetic compound (I-BET) that by ‘mimicking’ acetylated histones disrupts chromatin complexes responsible for the expression of key inflammatory genes in activated macrophages, and confers protection against lipopolysaccharide-induced endotoxic shock and bacteria-induced sepsis. Our findings suggest that synthetic compounds specifically targeting proteins that recognize post-translationally modified histones can serve as a new generation of immunomodulatory drugs.

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Accessions

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Gene Expression Omnibus

Protein Data Bank

Data deposits

Crystal structure of the first bromodomain of human BRD4 in complex with I-BET inhibitor was deposited in the RCSB Protein Data Bank with PDB ID code 3P5O. Microarray and ChIP sequencing results were deposited in GEO with GEO accession codes GSE21764 and GSE21910, respectively.

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Acknowledgements

We would like to acknowledge R. Grimley and C. Patel for supplying FRET data and R. Woodward, C. Delves, E. Jones and P. Holmes for protein production. J. Witherington, N. Smithers, S. Baddeley, J. Seal and L. Cutler provided compound selectivity and pharmacokinetics data. G. Krysa, O. Mirguet and R. Gosmini contributed to the discovery, development and characterization of the compound. We thank R. Anthony and S. McCleary for assistance with animal models, R. Gejman for bioinformatics analysis of gene expression kinetics and A. Santana and T. Chapman for technical assistance. We would like to thank C. Nathan, R. Medzhitov, S. Rudensky and S. Smale for helpful discussions and S. Sampath for his contribution to the concept of ‘histone mimicry’. R.C. is supported by an NIH KL2 Career Development Award and I.M. is supported by the American Italian Cancer Foundation. K.L.J. is supported by the National Health and Medical Research Council of Australia and is currently a Rockefeller University Women in Science Fellow.

Author information

Author notes

    • Edwige Nicodeme
    • , Kate L. Jeffrey
    • , Uwe Schaefer
    •  & Soren Beinke

    These authors contributed equally to this work.

Affiliations

  1. Centre de Recherche GSK, 27 Avenue du Québec, 91140 Villebon Sur Yvette, France

    • Edwige Nicodeme
    • , Hervé Coste
    •  & Jorge Kirilovsky
  2. Laboratory of Lymphocyte Signaling, The Rockefeller University, 1230 York Avenue, New York, New York 10065, USA

    • Kate L. Jeffrey
    • , Uwe Schaefer
    • , Rohit Chandwani
    • , Ivan Marazzi
    •  & Alexander Tarakhovsky
  3. Epinova DPU, Immuno-Inflammation Centre of Excellence for Drug Discovery, GlaxoSmithKline, Medicines Research Centre, Gunnels Wood Road, Stevenage SG1 2NY, UK

    • Soren Beinke
    • , Jose M. Lora
    • , Rab K. Prinjha
    •  & Kevin Lee
  4. Genomics Resource Center, The Rockefeller University, 1230 York Avenue, New York, New York 10065, USA

    • Scott Dewell
  5. GlaxoSmithKline R&D, Medicines Research Centre, Gunnels Wood Road, Stevenage SG1 2NY, UK

    • Chun-wa Chung
    • , Paul Wilson
    •  & Julia White
  6. Laboratory of Virology and Infectious Disease, The Rockefeller University, 1230 York Avenue, New York, New York 10065, USA

    • Charles M. Rice

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Contributions

E.N. identified, characterized and optimized the compound for in vivo experiments; K.L.J., U.S. and S.B. contributed equally to design, execution and analysis of in vitro and in vivo experiments. S.D. performed bioinformatics analysis of ChIP sequencing data; C.-w.C. performed crystallography, ITC, SPR and thermal shift assays; R.C. performed quantitative analysis of epigenetic states of the LPS-inducible genes; I.M. optimized BRD2 and BRD3 profiling of the LPS-inducible genes; P.W. performed bioinformatics analysis of gene expression in LPS-stimulated macrophages. H.C., J.W. and J.K. discovered, characterised and optimised the compound for in vivo experiments. C.M.R. was involved in studies of inflammatory responses. J.M.L., R.K.P. and K.L. contributed to the initiation and development of the studies on pharmacological targeting of proteins that recognize post-translationally modified histones. A.T. conceived and supervised this study, and wrote the final manuscript.

Competing interests

E.N., S.B., C.-w.C., P.W., H.C., J.W., J.K., J.M.L., R.K.P. and K.L. are employees of GlaxoSmithKline. Research support, excluding salaries to the members of The Rockefeller University, but including protein analysis and compound synthesizing equipment, supplies and other expense, was provided by GlaxoSmithKline.

Corresponding authors

Correspondence to Kevin Lee or Alexander Tarakhovsky.

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    Supplementary Information

    The file contains Supplementary Figures 1-14 with legends, Supplementary Tables 1-3, Supplementary Methods and additional references.

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https://doi.org/10.1038/nature09589

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