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Competitive blocking of LRP4–sclerostin binding interface strongly promotes bone anabolic functions

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Abstract

Induction of bone formation by Wnt ligands is inhibited when sclerostin (Scl), an osteocyte-produced antagonist, binds to its receptors, the low-density lipoprotein receptor-related proteins 5 or 6 (LRP5/6). Recently, it was shown that enhanced inhibition is achieved by Scl binding to the co-receptor LRP4. However, it is not clear if the binding of Scl to LRP4 facilitates Scl binding to LRP5/6 or inhibits the Wnt pathway in an LRP5/6-independent manner. Here, using the yeast display system, we demonstrate that Scl exhibits a stronger binding affinity for LRP4 than for LRP6. Moreover, we found stronger Scl binding to LRP6 in the presence of LRP4. We further show that a Scl mutant (SclN93A), which tightly binds LRP4 but not LRP6, does not inhibit the Wnt pathway on its own. We demonstrate that SclN93A competes with Scl for a common binding site on LRP4 and antagonizes Scl inhibition of the Wnt signaling pathway in osteoblasts in vitro. Finally, we demonstrate that 2 weeks of bi-weekly subcutaneous injections of SclN93A fused to the fragment crystallizable (Fc) domain of immunoglobulin (SclN93AFc), which retains the antagonistic activity of the mutant, significantly increases bone formation rate and enhances trabecular volumetric bone fraction, trabecular number, and bone length in developing mice. Our data show that LRP4 serves as an anchor that facilitates Scl–LRP6 binding and that inhibition of the Wnt pathway by Scl depends on its prior binding to LRP4. We further provide evidence that compounds that inhibit Scl–LRP4 interactions offer a potential strategy to promote anabolic bone functions.

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Acknowledgements

This work was supported by grants from the ISRAEL SCIENCE FOUNDATION no. 357/18 (NL) and no. 1615/19 (NP).

Funding

This work was supported by grants from the ISRAEL SCIENCE FOUNDATION no. 357/18 (NL) and no. 1615/19 (NP).

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Authors and Affiliations

  1. Department of Physiology and Cell Biology, Faculty of Health Sciences, Ben-Gurion University of the Negev, 8410501, Beer-Sheva, Israel

    Svetlana Katchkovsky, Biplab Chatterjee, Chen-Viki Abramovitch-Dahan & Noam Levaot

  2. Avram and Stella Goldstein-Goren Department of Biotechnology Engineering and the National Institute of Biotechnology in the Negev, Ben-Gurion University of the Negev, 8410501, Beer-Sheva, Israel

    Niv Papo

  3. Regenerative Medicine and Stem Cell Research Center, Ben-Gurion University of the Negev, 8410501, Beer-Sheva, Israel

    Noam Levaot

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  1. Svetlana Katchkovsky
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SK, conceptualization, data curation, investigation, methodology, validation, visualization, writing—original draft, writing—review and editing. BC, methodology. CA-D, methodology. NP, conceptualization, formal analysis, funding acquisition, investigation, methodology, resources, supervision, validation, visualization, writing—original draft, writing—review and editing. NL, conceptualization, formal analysis, funding acquisition, investigation, methodology, resources, supervision, validation, visualization, writing—original draft, writing—review and editing.

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Correspondence to Niv Papo or Noam Levaot.

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Katchkovsky, S., Chatterjee, B., Abramovitch-Dahan, CV. et al. Competitive blocking of LRP4–sclerostin binding interface strongly promotes bone anabolic functions. Cell. Mol. Life Sci. 79, 113 (2022). https://doi.org/10.1007/s00018-022-04127-2

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