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Persistent signaling induced by FTY720-phosphate is mediated by internalized S1P1 receptors

A Corrigendum to this article was published on 01 December 2009

This article has been updated

Abstract

Targeting sphingosine-1-phosphate receptors with the oral immunomodulator drug FTY720 (fingolimod) has demonstrated substantial efficacy in the treatment of multiple sclerosis. The drug is phosphorylated in vivo, and most of the clinical effects of FTY720-phosphate (FTY720P) are thought to be mediated via S1P1 receptors on lymphocytes and endothelial cells, leading to sequestration of lymphocytes in secondary lymphoid organs. FTY720P was described to act as a “functional antagonist” by promoting efficient internalization of S1P1 receptors. We demonstrate here that S1P1 receptors activated by FTY720P retain signaling activity for hours in spite of a quantitative internalization. Structural analogs of FTY720P with shorter alkyl side chains retained potency and efficacy in a functional assay but failed to promote long-lasting receptor internalization and signaling. We show that persistent signaling translates into an increased chemokinetic migration of primary human umbilical vein endothelial cells, which suggests persistent agonism as a crucial parameter in the mechanism of action of FTY720.

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Figure 1: Internalization and persistent signaling of S1P1 receptors in CHO-S1P1.
Figure 2: Structural determinants for persistent signaling of FTY720P-like agonists.
Figure 3: Effects of the S1P1 antagonist W146 on persistent signaling and internalization in CHO-S1P1.
Figure 4: Internalization and persistent signaling of endogenous S1P1 receptors in HUVECs.
Figure 5: Trafficking of S1P1 and migration of HUVECs.

Change history

  • 13 November 2009

    In the version of this article initially published, the chemical structures in Fig. 2a and in the graphical abstract were incorrect. The error has been corrected in the HTML and PDF versions of the article.

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Acknowledgements

We thank B. Wilmering-Wetter and S. Vidal for excellent technical assistance. We are also indebted to K. Hinterding and B. Bollbuck (Novartis Pharma AG) for the synthesis of tool compounds.

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Contributions

F.M. conceived, designed and performed experiments and wrote the manuscript. K.S. conceived and designed experiments and wrote the manuscript. F.Z. synthesized and analyzed chemical compounds. A.B. designed and performed experiments. C.C. designed and performed image analyses. D.G. designed and performed experiments.

Corresponding authors

Correspondence to Florian Mullershausen or Klaus Seuwen.

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Competing interests

F.M., F.Z., A.B., D.G. and K.S. are employees of Novartis Pharma AG.

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Mullershausen, F., Zecri, F., Cetin, C. et al. Persistent signaling induced by FTY720-phosphate is mediated by internalized S1P1 receptors. Nat Chem Biol 5, 428–434 (2009). https://doi.org/10.1038/nchembio.173

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