Cis-4-Methylsphingosine is a Sphingosine-1-phosphate Receptor Modulator

Michael ter Braak (1) , Ralf Frederik Claas (2) , Bianca Hegen (1) , Sandra Labocha (2) , Nerea Ferreirós (2) , Josef Pfeilschifter (2) , Andrea Huwiler (3) , Gerhild van Echten-Deckert (4) , Dagmar Meyer Zu Heringdorf (2)

1 Universitätsklinikum Essen
2 Klinikum der Goethe-Universität Frankfurt am Main
3 UNIBE - Universität Bern / University of Bern
4 Kekulé-Institut

Sphingosine-1-phosphate (S1P) acts as high affinity agonist at specific G-protein-coupled receptors, S1P, that play important roles e.g. in the cardiovascular and immune systems. A S1P receptor modulating drug, FTY720 (fingolimod), has been effective in phase III clinical trials for multiple sclerosis. FTY720 is a sphingosine analogue and prodrug of FTY720-phosphate, which activates all S1P receptors except S1P and disrupts lymphocyte trafficking by internalizing the S1P receptor. Cis-4-methylsphingosine (cis-4M-Sph) is another synthetic sphingosine analogue that is readily taken up by cells and phosphorylated to cis-4-methylsphingosine-1-phosphate (cis-4M-S1P). Therefore, we analysed whether cis-4M-Sph interacted with S1P receptors through its metabolite cis-4M-S1P in a manner similar to FTY720. Indeed, cis-4M-Sph caused an internalization of S1P receptors, but differed from FTY720 as it acted on S1P and S1P and only weakly on S1P, while FTY720 internalized S1P and S1P but not S1P. Consequently, pre-incubation with cis-4M-Sph specifically desensitized S1P-induced [Ca] increases, which are mediated by S1P and S1P, in a time- and concentration-dependent manner. This effect was not shared by sphingosine or FTY720, indicating that metabolic stability and targeting of S1P receptors were important. The desensitization of S1P-induced [Ca] increases was dependent on the expression of SphKs, predominantly of SphK2, and thus mediated by cis-4M-S1P. In agreement, cis-4M-S1P was detected in the supernatants of cells exposed to cis-4M-Sph. It is concluded that cis-4M-Sph, through its metabolite cis-4M-S1P, acts as a S1P receptor modulator and causes S1P receptor internalization and desensitization. The data furthermore help to define requirements for sphingosine kinase substrates as S1P receptor modulating prodrugs.

Mots clés: Sphingosine-1-phosphate; lysophospholipid receptors; GPCR; sphingosine kinase; FTY720

Michael ter Braak, Ralf Frederik Claas, Bianca Hegen, Sandra Labocha, Nerea Ferreirós, et al.. Cis-4-Methylsphingosine is a Sphingosine-1-phosphate Receptor Modulator. Biochemical Pharmacology, 2011, 81 (5), pp.617. ⟨10.1016/j.bcp.2010.12.002⟩. ⟨hal-00663402⟩