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.
Subscribe to Journal
Get full journal access for 1 year
only $9.92 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
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.
Straub, R. H. & Schradin, C. Chronic inflammatory systemic diseases: An evolutionary trade-off between acutely beneficial but chronically harmful programs. Evol. Med. Public Health 2016, 37–51 (2016).
Harrington, L. E. et al. Interleukin 17-producing CD4+ effector T cells develop via a lineage distinct from the T helper type 1 and 2 lineages. Nat. Immunol. 6, 1123–1132 (2005).
Park, H. et al. A distinct lineage of CD4 T cells regulates tissue inflammation by producing interleukin 17. Nat. Immunol. 6, 1133–1141 (2005).
Mohan, C. & Putterman, C. Genetics and pathogenesis of systemic lupus erythematosus and lupus nephritis. Nat. Rev. Nephrol. 11, 329–341 (2015).
Chiche, L. et al. New treatment options for lupus - a focus on belimumab. Ther. Clin. Risk Manag. 8, 33–43 (2012).
Tauzin, S. et al. The naturally processed CD95L elicits a c-yes/calcium/PI3K-driven cell migration pathway. PLoS Biol. 9, e1001090 (2011).
Suda, T., Takahashi, T., Golstein, P. & Nagata, S. Molecular cloning and expression of the Fas ligand, a novel member of the tumor necrosis factor family. Cell 75, 1169–1178 (1993).
O’ Reilly, L. A. et al. Membrane-bound Fas ligand only is essential for Fas-induced apoptosis. Nature 461, 659–663 (2009).
O’Reilly, K. E. et al. mTOR inhibition induces upstream receptor tyrosine kinase signaling and activates Akt. Cancer Res. 66, 1500–1508 (2006).
Holler, N. et al. Two adjacent trimeric Fas ligands are required for Fas signaling and formation of a death-inducing signaling complex. Mol. Cell. Biol. 23, 1428–1440 (2003).
Kischkel, F. C. et al. Cytotoxicity-dependent APO-1 (Fas/CD95)-associated proteins form a death-inducing signaling complex (DISC) with the receptor. EMBO J. 14, 5579–5588 (1995).
Kleber, S. et al. Yes and PI3K bind CD95 to signal invasion of glioblastoma. Cancer Cell 13, 235–248 (2008).
Malleter, M. et al. CD95L cell surface cleavage triggers a prometastatic signaling pathway in triple-negative breast cancer. Cancer Res. 73, 6711–6721 (2013).
Poissonnier, A. et al. CD95-mediated calcium signaling promotes T helper 17 trafficking to inflamed organs in lupus-prone mice. Immunity 45, 209–223 (2016).
Strasser, A., Jost, P. J. & Nagata, S. The many roles of FAS receptor signaling in the immune system. Immunity 30, 180–192 (2009).
Bechara, C. & Sagan, S. Cell-penetrating peptides: 20 years later, where do we stand? FEBS Lett. 587, 1693–1702 (2013).
Gokhale, A. S. & Satyanarayanajois, S. Peptides and peptidomimetics as immunomodulators. Immunotherapy 6, 755–774 (2014).
Hu, Y., Li, X., Sebti, S. M., Chen, J. & Cai, J. Design and synthesis of AApeptides: a new class of peptide mimics. Bioorg. Med. Chem. Lett. 21, 1469–1471 (2011).
Deng, L., Velikovsky, C. A., Swaminathan, C. P., Cho, S. & Mariuzza, R. A. Structural basis for recognition of the T cell adaptor protein SLP-76 by the SH3 domain of phospholipase Cgamma1. J. Mol. Biol. 352, 1–10 (2005).
Berry, D. M., Nash, P., Liu, S. K., Pawson, T. & McGlade, C. J. A high-affinity Arg-X-X-Lys SH3 binding motif confers specificity for the interaction between Gads and SLP-76 in T cell signaling. Curr. Biol. 12, 1336–1341 (2002).
Sanzenbacher, R., Kabelitz, D. & Janssen, O. SLP-76 binding to p56lck: a role for SLP-76 in CD4-induced desensitization of the TCR/CD3 signaling complex. J. Immunol. 163, 3143–3152 (1999).
Yablonski, D., Kadlecek, T. & Weiss, A. Identification of a phospholipase C-gamma1 (PLC-gamma1) SH3 domain-binding site in SLP-76 required for T-cell receptor-mediated activation of PLC-gamma1 and NFAT. Mol. Cell. Biol. 21, 4208–4218 (2001).
Harkiolaki, M. et al. Structural basis for SH3 domain-mediated high-affinity binding between Mona/Gads and SLP-76. EMBO J. 22, 2571–2582 (2003).
Clements, J. L. et al. Requirement for the leukocyte-specific adapter protein SLP-76 for normal T cell development. Science 281, 416–419 (1998).
Pivniouk, V. et al. Impaired viability and profound block in thymocyte development in mice lacking the adaptor protein SLP-76. Cell 94, 229–238 (1998).
Yablonski, D., Kuhne, M. R., Kadlecek, T. & Weiss, A. Uncoupling of nonreceptor tyrosine kinases from PLC-gamma1 in an SLP-76-deficient T cell. Science 281, 413–416 (1998).
Bénéteau, M. et al. Localization of Fas/CD95 into the lipid rafts on down-modulation of the phosphatidylinositol 3-kinase signaling pathway. Mol. Cancer Res. 6, 604–613 (2008).
Straus, S. E. et al. The development of lymphomas in families with autoimmune lymphoproliferative syndrome with germline Fas mutations and defective lymphocyte apoptosis. Blood 98, 194–200 (2001).
Adachi, M., Watanabe-Fukunaga, R. & Nagata, S. Aberrant transcription caused by the insertion of an early transposable element in an intron of the Fas antigen gene of lpr mice. Proc. Natl. Acad. Sci. USA 90, 1756–1760 (1993).
Theofilopoulos, A. N. & Dixon, F. J. Murine models of systemic lupus erythematosus. Adv. Immunol. 37, 269–390 (1985).
Henderson, L. E. et al. Molecular characterization of gag proteins from simian immunodeficiency virus (SIVMne). J. Virol. 62, 2587–2595 (1988).
Rich, D. H., Green, J., Toth, M. V., Marshall, G. R. & Kent, S. B. Hydroxyethylamine analogues of the p17/p24 substrate cleavage site are tight-binding inhibitors of HIV protease. J. Med. Chem. 33, 1285–1288 (1990).
Erickson, J. et al. Design, activity, and 2.8 A crystal structure of a C2 symmetric inhibitor complexed to HIV-1 protease. Science 249, 527–533 (1990).
Flexner, C. HIV-protease inhibitors. N. Engl. J. Med. 338, 1281–1292 (1998).
Rubino, S. J., Geddes, K. & Girardin, S. E. Innate IL-17 and IL-22 responses to enteric bacterial pathogens. Trends Immunol. 33, 112–118 (2012).
Alfonso, Y. & Monzote, L. HIV protease inhibitors: effect on the opportunistic protozoan parasites. Open Med. Chem. J. 5, 40–50 (2011).
Mody, G. M., Patel, N., Budhoo, A. & Dubula, T. Concomitant systemic lupus erythematosus and HIV: case series and literature review. Semin. Arthritis Rheum. 44, 186–194 (2014).
Langley, R. G. et al. Secukinumab in plaque psoriasis–results of two phase 3 trials. N. Engl. J. Med. 371, 326–338 (2014).
Leonardi, C. et al. Anti-interleukin-17 monoclonal antibody ixekizumab in chronic plaque psoriasis. N. Engl. J. Med. 366, 1190–1199 (2012).
Papp, K. A. et al. Brodalumab, an anti-interleukin-17-receptor antibody for psoriasis. N. Engl. J. Med. 366, 1181–1189 (2012).
Berthelot, P., Guglielminotti, C., Frésard, A., Lucht, F. & Perrot, J. L. Dramatic cutaneous psoriasis improvement in a patient with the human immunodeficiency virus treated with 2′,3′-dideoxy,3′-thyacytidine [correction of 2′,3′-dideoxycytidine] and ritonavir. Arch. Dermatol 133, 531 (1997).
Chiricozzi, A. et al. Complete resolution of erythrodermic psoriasis in an HIV and HCV patient unresponsive to antipsoriatic treatments after highly active antiretroviral therapy (ritonavir, atazanavir, emtricitabine, tenofovir). Dermatology 225, 333–337 (2012).
Fischer, T., Schwörer, H., Vente, C., Reich, K. & Ramadori, G. Clinical improvement of HIV-associated psoriasis parallels a reduction of HIV viral load induced by effective antiretroviral therapy. AIDS 13, 628–629 (1999).
Abramoff, M. D., Magelhaes, P. J. & Ram, S. J. Image processing with Image. J. Biophoton. Int. 11, 36–42 (2004).
Marinozzi, V. New technics for staining tissues embedded in plastic material for study with the high resolution light microscope. Z. Wiss. Mikrosk. 65, 219–230 (1963).
Jiang, C. et al. Abrogation of lupus nephritis in activation-induced deaminase-deficient MRL/lpr mice. J. Immunol. 178, 7422–7431 (2007).
Legembre, P., Moreau, P., Daburon, S., Moreau, J. F. & Taupin, J. L. Potentiation of Fas-mediated apoptosis by an engineered glycosylphosphatidylinositol-linked Fas. Cell Death Differ. 9, 329–339 (2002).
Delaglio, F. et al. NMRPipe: a multidimensional spectral processing system based on UNIX pipes. J. Biomol. NMR 6, 277–293 (1995).
Lee, W., Tonelli, M. & Markley, J. L. NMRFAM-SPARKY: enhanced software for biomolecular NMR spectroscopy. Bioinformatics 31, 1325–1327 (2015).
Padhee, S. et al. Non-hemolytic α-AApeptides as antimicrobial peptidomimetics. Chem. Commun. (Camb.) 47, 9729–9731 (2011).
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.).
The authors declare no competing interests.
Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
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
Clinical Reviews in Allergy & Immunology (2020)