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  • A Correction to this article was published on 04 May 2018

Abstract

Regeneration of the adult intestinal epithelium is mediated by a pool of cycling stem cells, which are located at the base of the crypt, that express leucine-rich-repeat-containing G-protein-coupled receptor 5 (LGR5). The Frizzled (FZD) 7 receptor (FZD7) is enriched in LGR5+ intestinal stem cells and plays a critical role in their self-renewal. Yet, drug discovery approaches and structural bases for targeting specific FZD isoforms remain poorly defined. FZD proteins interact with Wnt signaling proteins via, in part, a lipid-binding groove on the extracellular cysteine-rich domain (CRD) of the FZD receptor. Here we report the identification of a potent peptide that selectively binds to the FZD7 CRD at a previously uncharacterized site and alters the conformation of the CRD and the architecture of its lipid-binding groove. Treatment with the FZD7-binding peptide impaired Wnt signaling in cultured cells and stem cell function in intestinal organoids. Together, our data illustrate that targeting the lipid-binding groove holds promise as an approach for achieving isoform-selective FZD receptor inhibition.

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Change history

  • 04 May 2018

    The version of this article originally published contained older versions of the Life Sciences Reporting Summary and the Supplementary Text and Figures. The error has been corrected in the HTML and PDF versions of the article.

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Acknowledgements

We thank R. Ferrao for assistance with collecting the crystallographic dataset, S. Gierke for help with microscopy, A. Estevez, C. Ciferri, K. Mortara and the Baculovirus expression group for assistance with protein expression, R. Ferrao and D. Whalen for helpful discussions, and the Genentech FACS core for technical assistance. Molecular graphics and analyses were performed with the UCSF Chimera package. Chimera was developed by the Resource for Biocomputing, Visualization and Informatics at the University of California, San Francisco (supported by US National Institutes of Health (NIH)-NIGMS grant no. P41-GM103311).

Author information

Affiliations

  1. Department of Early Discovery Biochemistry, Genentech, South San Francisco, CA, USA

    • Aaron H. Nile
    • , Simon Hansen
    • , Lijuan Zhou
    • , Yingnan Zhang
    • , Yue Fu
    • , Emily B. Gogol
    • , Wayne J. Fairbrother
    •  & Rami N. Hannoush
  2. Department of Molecular Oncology, Genentech, South San Francisco, CA, USA

    • Felipe de Sousa e Melo
    •  & Frederic J. de Sauvage
  3. Department of Structural Biology, Genentech, South San Francisco, CA, USA

    • Susmith Mukund
    • , Christopher Koth
    •  & Weiru Wang
  4. Department of Bioinformatics and Computational Biology, Genentech, South San Francisco, CA, USA

    • Robert Piskol
  5. Department of Pathology, Genentech, South San Francisco, CA, USA

    • László G. Kömüves
  6. Department of Molecular Biology, Genentech, South San Francisco, CA, USA

    • Zora Modrusan
  7. Department of Pharmaceutical Sciences, University of Toronto, Toronto, Ontario, Canada

    • Stephane Angers
  8. Department of Biomolecular Resources, Genentech, South San Francisco, CA, USA

    • Yvonne Franke

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Contributions

A.H.N., F.d.S.e.M., S.M., R.P., S.H., L.Z., Y.Z., Y.F., E.B.G., Z.M., S.A., Y.F., C.K., W.J.F., W.W., F.J.d.S. and R.N.H. designed research; A.H.N. performed protein purification and characterization, and biochemical and cellular assays; F.d.S.e.M. performed organoid assays; S.M. performed crystallography experiments; S.H. performed SPR experiments; L.Z. performed phage panning; Y.F. designed expression constructs; E.B.G. and R.N.H. performed cellular assays; L.G.K. performed confocal microscopy; A.H.N. and W.W. solved the X-ray crystal structure; R.P. analyzed the RNA-seq data; A.H.N. and R.N.H. wrote the manuscript, with input from the other authors; and R.N.H. conceived and directed the study.

Competing interests

The authors are employees of Genentech, a member of the Roche group. A patent (PCT/US2017/050841; A.H.N., Y.Z., L.Z., and R.N.H.) covering this work is pending.

Corresponding author

Correspondence to Rami N. Hannoush.

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    Supplementary Tables 1–4 and Supplementary Figures 1–25

  2. Life Sciences Reporting Summary

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https://doi.org/10.1038/s41589-018-0035-2