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Inhibition of Delta-induced Notch signaling using fucose analogs

Nature Chemical Biology volume 14, pages 65–71 (2018)Cite this article

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Abstract

Notch is a cell-surface receptor that controls cell-fate decisions and is regulated by O-glycans attached to epidermal growth factor-like (EGF) repeats in its extracellular domain. Protein O-fucosyltransferase 1 (Pofut1) modifies EGF repeats with O-fucose and is essential for Notch signaling. Constitutive activation of Notch signaling has been associated with a variety of human malignancies. Therefore, tools that inhibit Notch activity are being developed as cancer therapeutics. To this end, we screened L-fucose analogs for their effects on Notch signaling. Two analogs, 6-alkynyl and 6-alkenyl fucose, were substrates of Pofut1 and were incorporated directly into Notch EGF repeats in cells. Both analogs were potent inhibitors of binding to and activation of Notch1 by Notch ligands Dll1 and Dll4, but not by Jag1. Mutagenesis and modeling studies suggest that incorporation of the analogs into EGF8 of Notch1 markedly reduces the ability of Delta ligands to bind and activate Notch1.

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Figure 1: Effects of fucose analogs on Notch signaling in Zebrafish embryos.
Figure 2: Peracetylated fucose analogs are efficiently incorporated into Notch EGF repeats and elongated by Lfng.
Figure 3: Peracetylated fucose analogs inhibit Dll1- and Dll4- but not Jag1-induced Notch signaling.
Figure 4: Compounds 10 and 11 inhibit Notch–Dll ligand binding.
Figure 5: Incorporation of fucose analogs at EGF8 of Notch1 plays an important role in Dll1- and Dll4-mediated Notch activation.
Figure 6: Fucose analog 10 inhibits the development of T-cell progenitors.

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Acknowledgements

This work was supported by NIH grants R01GM061126 (R.S.H.), R01GM093282 (P.W.), R01GM106417 (P.S.) and K99CA204738 (V.C.L.). M.S. was partially supported by T32GM00844. We thank the following for providing materials: J. Nye (Northwestern University Medical School), S. Chiba (University of Tokyo), G. Bornkamm (Helmholtz Zentrum Munchen), G. Weinmater (UCLA), S. Blacklow (Harvard), S. Kakuda (Stony Brook University), and C. Guidos (University of Toronto).

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Author notes

  1. Vincent C Luca

    Present address: Department of Drug Discovery, H. Lee Moffitt Cancer Center and Research Institute, Tampa, Florida, USA

  2. Vivek Kumar and Lars Ulrik Nordstrøm: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biochemistry and Cell Biology, Stony Brook University, Stony Brook, New York, USA

    Michael Schneider, Hideyuki Takeuchi & Robert S Haltiwanger

  2. Department of Cell Biology, Albert Einstein College of Medicine, New York, USA

    Vivek Kumar & Pamela Stanley

  3. Department of Biochemistry, Albert Einstein College of Medicine, New York, USA

    Lars Ulrik Nordstrøm, Lei Feng & Peng Wu

  4. Complex Carbohydrate Research Center, University of Georgia, Athens, Georgia, USA

    Hideyuki Takeuchi, Huilin Hao & Robert S Haltiwanger

  5. Departments of Molecular and Cellular Physiology and Structural Biology, Stanford University School of Medicine, Stanford, California, USA

    Vincent C Luca & K Christopher Garcia

  6. Howard Hughes Medical Institute, Stanford, California, USA

    Vincent C Luca & K Christopher Garcia

  7. Department of Molecular Medicine, The Scripps Research Institute, La Jolla, California, USA

    Peng Wu

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Contributions

M.S., P.W. and R.S.H. conceived experiments, analyzed and interpreted data and wrote the paper. L.U.N. synthesized fucose analogs. M.S., V.K., L.F., H.T. and H.H. designed and performed experiments. V.C.L. and K.C.G. constructed structural models. P.S. and P.W. helped design experiments, interpret data and revise the manuscript.

Corresponding authors

Correspondence to Peng Wu or Robert S Haltiwanger.

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The authors declare no competing financial interests.

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Schneider, M., Kumar, V., Nordstrøm, L. et al. Inhibition of Delta-induced Notch signaling using fucose analogs. Nat Chem Biol 14, 65–71 (2018). https://doi.org/10.1038/nchembio.2520

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