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A selective inhibitor reveals PI3Kγ dependence of TH17 cell differentiation

Nature Chemical Biology volume 8, pages 576–582 (2012)Cite this article

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A Corrigendum to this article was published on 18 July 2012

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Abstract

We devised a high-throughput chemoproteomics method that enabled multiplexed screening of 16,000 compounds against native protein and lipid kinases in cell extracts. Optimization of one chemical series resulted in CZC24832, which is to our knowledge the first selective inhibitor of phosphoinositide 3-kinase γ (PI3Kγ) with efficacy in in vitro and in vivo models of inflammation. Extensive target- and cell-based profiling of CZC24832 revealed regulation of interleukin-17–producing T helper cell (TH17) differentiation by PI3Kγ, thus reinforcing selective inhibition of PI3Kγ as a potential treatment for inflammatory and autoimmune diseases.

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Figure 1: Chemoproteomic profiling of PI3K inhibitors.
Figure 2: Design and characterization of CZC24832, a selective PI3Kγ inhibitor.
Figure 3: CZC24832 reveals a new role of PI3Kγ in TH17 regulation.

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  • 16 May 2012

    In the version of this article initially published, the name M. Sunose was misspelled in the Acknowledgements. The error has been corrected in the HTML and PDF versions of the article.

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Acknowledgements

We are grateful to M. Sunhose and W. Miller for chemistry and cheminformatics support; MS, information technology, biochemistry and biology groups for technical support; G. Creighton-Gutteridge for experimental support; F. Weisbrodt for assistance with graphics; G. Bennett (Cellzome Ltd.) and O. Azzolino (University of Torino) for animal data; Y. Abraham for generation of the cladogram; R. Hale and T. Edwards for general advice; and M. Wymann, D. Simmons and A. Watt for stimulating discussions and valuable comments.

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Authors and Affiliations

  1. Cellzome AG, Heidelberg, Germany

    Giovanna Bergamini, Satoko Shimamura, Thilo Werner, Katrin Müller, Jessica Perrin, Christina Rau, Carsten Hopf, Carola Doce, Toby Mathieson, Faiza Rharbaoui, Friedrich Reinhard, Mikhail M Savitski, Gerard Drewes, Marcus Bantscheff & Gitte Neubauer

  2. Cellzome Ltd., Chesterford Research Park, Cambridge, UK.,

    Kathryn Bell, Andrew Cansfield, Katie Ellard, Daniel Leggate, Raffaella Mangano, Nigel Ramsden & Oliver Rausch

  3. BioSeek LLC, San Francisco, California, USA

    Alison O'Mahony & Ivan Plavec

  4. Department of Genetics, Biology and Biochemistry, Molecular Biotechnology Center, University of Torino, Torino, Italy

    Emilio Hirsch

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Contributions

G.B., S.S., K.M. and J.P. developed and performed assays; K.B., A.C. and K.E. designed and synthesized compounds; C.R., F. Reinhard, D.L. and R.M. performed screening; F. Rharbaoui contributed to animal studies; M.M.S., T.M. and M.B. developed MS technology; C.D. supported data analysis and display; A.O. and I.P. contributed cellular activity profiles; T.W. designed and performed MS experiments; C.H. supervised biochemistry work; G.D. contributed to the manuscript and gave advice; E.H. gave general advice and contributed animal data; N.R. led the chemistry efforts; O.R. contributed to data analysis and manuscript preparation; and G.B., M.B. and G.N. designed and supervised the study, analyzed data and wrote the manuscript.

Corresponding authors

Correspondence to Marcus Bantscheff or Gitte Neubauer.

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Competing interests

G.B., S.S., T.W., K.M., J.P., C.R., A.H., C.D., T.M., F. Rharbaoui, F. Reinhard, M.M.S., G.D., M.B. and G.N. are employees of Cellzome AG; K.B., A.C., K.E., D.L., R.M., N.R. and O.R. are employees of Cellzome Ltd.; and A.M. and I.P. are employees of BioSeek Inc.

Supplementary information

Supplementary Text and Figures

Supplementary Methods and Supplementary Results (PDF 11964 kb)

Supplementary Data Set 1

Complementary kinase capturing of CZC15098 and CZC15292 derived bead matrices in different cell extracts (XLSX 256 kb)

Supplementary Data Set 2

Concentration-inhibition profiles for all reference compounds and CZC19945 and CZC24832 in different cell extracts (XLSX 902 kb)

Supplementary Data Set 3

Correction factors for calculation of apparent dissociation constants from IC50s obtained in profiling experiments using kinobeads and HL60 cell extracts (XLSX 175 kb)

Supplementary Data Set 4

Correction factors for calculation of apparent dissociation constants from IC50s obtained in profiling experiments using LK beads and HL60 cell extracts (XLSX 163 kb)

Supplementary Data Set 5

Proteomic binding profiles using an immobilized linkable analogue of CZC19945/CZC24832 as probe matrix and HeLa, HL60, and PBMC cell extracts (XLSX 175 kb)

Supplementary Data Set 6

Correction factors for calculation of apparent dissociation constants from IC50s obtained in profiling experiments using LK beads and mixed raw264.7, HL60 cell extracts (XLSX 163 kb)

Supplementary Data Set 7

Correction factors for calculation of apparent dissociation constants from IC50s obtained in profiling experiments using LK beads and mixed RBL-1, HL60 cell extracts (XLSX 163 kb)

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Bergamini, G., Bell, K., Shimamura, S. et al. A selective inhibitor reveals PI3Kγ dependence of TH17 cell differentiation. Nat Chem Biol 8, 576–582 (2012). https://doi.org/10.1038/nchembio.957

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