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Disrupting the CD95–PLCγ1 interaction prevents Th17-driven inflammation

Nature Chemical Biology volume 14, pages 1079–1089 (2018)Cite this article

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Abstract

CD95L is a transmembrane ligand (m-CD95L) that is cleaved by metalloproteases to release a soluble ligand (s-CD95L). Unlike m-CD95L, interaction between s-CD95L and CD95 fails to recruit caspase-8 and FADD to trigger apoptosis and instead induces a Ca2+ response via docking of PLCγ1 to the calcium-inducing domain (CID) within CD95. This signaling pathway induces accumulation of inflammatory Th17 cells in damaged organs of lupus patients, thereby aggravating disease pathology. A large-scale screen revealed that the HIV protease inhibitor ritonavir is a potent disruptor of the CD95–PLCγ1 interaction. A structure–activity relationship approach highlighted that ritonavir is a peptidomimetic that shares structural characteristics with CID with respect to docking to PLCγ1. Thus, we synthesized CID peptidomimetics abrogating both the CD95-driven Ca2+ response and transmigration of Th17 cells. Injection of ritonavir and the CID peptidomimetic into lupus mice alleviated clinical symptoms, opening a new avenue for the generation of drugs for lupus patients.

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Fig. 1: Screening of the Prestwick library to identify chemical leads that inhibit the CD95–PLCγ1 interaction.
Fig. 2: Peptidomimetics inhibit CD95-mediated PLCγ1 recruitment, calcium signaling, and endothelial transmigration of Th17 T cells.
Fig. 3: NMR characterization of SLP-76, CID and DB550 binding surfaces to the SH3 domain of PLCγ1.
Fig. 4: Competitive effect of DB550 and ritonavir toward the SLP-76–PLCγ1 interaction.
Fig. 5: Ritonavir and DB550 alleviate clinical symptoms in lupus-prone mice.
Fig. 6: HIV protease inhibitors are responsible for inhibiting the CD95-mediated Ca2+ signaling pathway in cells isolated from HIV patients.

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Data availability

The authors declare that the data supporting the findings of this study are available within the paper and its supplementary information or are available from the corresponding author on reasonable request. Additional modeling methods are described Methods.

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Acknowledgements

We are grateful to the H2P2 facility at Biosit (Rennes) for its technical assistance and to M. Katan (Chester Beatty Laboratories, The Institute of Cancer Research, London, UK) and S.W. Michnick (University of Montréal, Canada) for providing vectors. This work was supported by INCa PLBIO (P.L., P.V., P.v.d.W. and M.J.), Ligue Contre le Cancer (PL), Fondation ARC (P.L.), ANR PRCE (P.L. and P.B.), Fondation Arthritis (P.B.) and Canadian Institutes of Health Research Grant FRN-156276 (K.G.).

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Author notes

  1. Daniel Best

    Present address: Sygnature Discovery, Nottingham, UK

  2. These authors contributed equally: Amanda Poissonnier, Jean-Philippe Guégan, Ha Thanh Nguyen.

Authors and Affiliations

  1. CLCC Eugène Marquis, Equipe Ligue Contre Le Cancer, Rennes, France

    Amanda Poissonnier, Jean-Philippe Guégan, Ha Thanh Nguyen, Daniel Best, Raphael Pineau, Florence Jouan, Lucie Morere, Sophie Martin, Mélissa Thomas, Pierre van de Weghe, Mickael Jean & Patrick Legembre

  2. Université Rennes, INSERM, UMR1242, Rennes, France

    Amanda Poissonnier, Jean-Philippe Guégan, Ha Thanh Nguyen, Daniel Best, Raphael Pineau, Florence Jouan, Lucie Morere, Sophie Martin, Mélissa Thomas, Pierre van de Weghe, Mickael Jean & Patrick Legembre

  3. Bioprojet Biotech, Saint-Grégoire, France

    Nicolas Levoin

  4. Department of Biochemistry, McGill Centre for Structural Biology, McGill University, Montreal, Canada

    Guennadi Kozlov, Kalle Gehring & Isabelle Douchet

  5. Université de Bordeaux, Bordeaux, France

    Estibaliz Lazaro, Thomas Ducret, Patrick Blanco & Pierre Vacher

  6. CNRS UMR 5164, Immunoconcept, Bordeaux, France

    Estibaliz Lazaro, Isabelle Douchet & Patrick Blanco

  7. CHU de Bordeaux, Bordeaux, France

    Estibaliz Lazaro & Patrick Blanco

  8. INSERM 1045, CRCTB, Bordeaux, France

    Thomas Ducret

  9. INSERM U1218, Bordeaux, France

    Pierre Vacher

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Contributions

A.P., J.-P.G., H.T.N., D.B., N.L., M.J., P.V., G.K., M.J., F.J., R.P., T.D., I.D., S.M., M.T., E.L. and L.M. conducted the experiments. N.L. developed the computer analyses. A.P., D.B., J.-P.G., N.L., P.B., G.K., K.G., P.V., M.J., P.v.d.W. and P.L. designed the experiments, analyzed data and wrote the paper. P.L. supervised the project.

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Correspondence to Patrick Legembre.

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Supplementary Information

Supplementary Figures 1–11, Supplementary Tables 1–6

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Synthetic procedures

Supplementary Dataset 1

NMR spectra raw data

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Poissonnier, A., Guégan, JP., Nguyen, H.T. et al. Disrupting the CD95–PLCγ1 interaction prevents Th17-driven inflammation. Nat Chem Biol 14, 1079–1089 (2018). https://doi.org/10.1038/s41589-018-0162-9

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