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WNT16 Requires Gα Subunits as Intracellular Partners for Both Its Canonical and Non-Canonical WNT Signalling Activity in Osteoblasts

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Abstract

In the past years, WNT16 became an interesting target in the field of skeletal research, as it was identified as an essential regulator of the cortical bone compartment, with the ability to increase both cortical and trabecular bone mass and strength in vivo. Even though there are indications that these advantageous effects are coming from canonical and non-canonical WNT-signalling activity, a clear model of WNT signalling by WNT16 is not yet depicted. We, therefore, investigated the modulation of canonical (WNT/β-catenin) and non-canonical [WNT/calcium, WNT/planar cell polarity (PCP)] signalling in human embryonic kidney (HEK) 293 T and SaOS2 cells. Here, we demonstrated that WNT16 activates all WNT-signalling pathways in osteoblasts, whereas only WNT/calcium signalling was activated in HEK293T cells. In osteoblasts, we therefore, additionally investigated the role of Gα subunits as intracellular partners in WNT16′s mechanism of action by performing knockdown of Gα12, Gα13 and Gαq. These studies point out that the above-mentioned Gα subunits might be involved in the WNT/β-catenin and WNT/calcium-signalling activity by WNT16 in osteoblasts, and for Gα12 in its WNT/PCP-signalling activity, illustrating a novel possible mechanism of interplay between the different WNT-signalling pathways in osteoblasts. Additional studies are needed to demonstrate whether this mechanism is specific for WNT16 signalling or relevant for all other WNT ligands as well. Altogether, we further defined WNT16′s mechanism of action in osteoblasts that might underlie the well-known beneficial effects of WNT16 on skeletal homeostasis. These findings on WNT16 and the activity of specific Gα subunits in osteoblasts could definitely contribute to the development of novel therapeutic approaches for fragility fractures in the future.

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Acknowledgements

This work was supported by the Systems Biology for the Functional Validation of Genetic Determinants of Skeletal Diseases (SYBIL, Project ID 602300) Project funded by the European Commission. G.H. holds a Rosa Blanckaert Legacy Grant for Young Investigators from the University of Antwerp (Project ID 32435; www.uantwerpen.be). E.B. holds a Post-doctoral Fellowship of the FWO Flanders (FWO Personal Grant 12A3814N). M.V. is supported by Donations from the “Siebe Van Reusel Fonds”.

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Author notes

  1. Gretl Hendrickx

    Present address: Department of Osteology and Biomechanics, University Medical Center Hamburg-Eppendorf, Hamburg, Germany

Authors and Affiliations

  1. Department of Medical Genetics, University of Antwerp, Antwerp, Belgium

    Gretl Hendrickx, Eveline Boudin, Marinus Verbeek, Erik Fransen, Geert Mortier & Wim Van Hul

  2. Department of Medical Genetics, University of Antwerp, Prins Boudewijnlaan 43, Edegem, 2650, Antwerp, Belgium

    Wim Van Hul

Authors
  1. Gretl Hendrickx
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  2. Eveline Boudin
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  3. Marinus Verbeek
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  4. Erik Fransen
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Contributions

Study design: GH, EB and WVH. Study conduct: GH. Data analysis: GH, EB, MV and EF. Data interpretation: all authors. Drafting manuscript: GH and MV. Revising manuscript: all authors. Approving manuscript for publication: all authors. GH takes responsibility for the integrity of the data analysis.

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Correspondence to Wim Van Hul.

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Gretl Hendrickx, Eveline Boudin, Marinus Verbeek, Erik Fransen, Geert Mortier and Wim Van Hul declare that they have no conflict of interest.

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Hendrickx, G., Boudin, E., Verbeek, M. et al. WNT16 Requires Gα Subunits as Intracellular Partners for Both Its Canonical and Non-Canonical WNT Signalling Activity in Osteoblasts. Calcif Tissue Int 106, 294–302 (2020). https://doi.org/10.1007/s00223-019-00633-x

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