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VWC2 Increases Bone Formation Through Inhibiting Activin Signaling

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Abstract

By a bioinformatics approach, we have identified a novel cysteine knot protein member, VWC2 (von Willebrand factor C domain containing 2) previously known as Brorin. Since Brorin has been proposed to function as a bone morphogenetic protein (BMP) antagonist, we investigated the binding of Brorin/VWC2 to several BMPs; however, none of the BMPs tested were bound to VWC2. Instead, the βA subunit of activin was found as a binding partner among transforming growth factor (TGF)-β superfamily members. Here, we show that Vwc2 gene expression is temporally upregulated early in osteoblast differentiation, VWC2 protein is present in bone matrix, and localized at osteoblasts/osteocytes. Activin A-induced Smad2 phosphorylation was inhibited in the presence of exogenous VWC2 in MC3T3-E1 osteoblast cell line and primary osteoblasts. The effect of VWC2 on ex vivo cranial bone organ cultures treated with activin A was investigated, and bone morphometric parameters decreased by activin A were restored with VWC2. When we further investigated the biological mechanism how VWC2 inhibited the effects of activin A on bone formation, we found that the effects of activin A on osteoblast cell growth, differentiation, and mineralization were reversed by VWC2. Taken together, a novel secretory protein, VWC2 promotes bone formation by inhibiting Activin-Smad2 signaling pathway.

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Acknowledgements

We thank Dr. Philip Trackman (Boston University) for his valuable comments on the manuscript. This study was supported by grants from the NIH (NIDCR; DE019527 and NIAMS; AR057451 to Y.Mo., NIDCR; DE020843 to Y.Mi.). The nucleotide sequences for mouse Vwc2 gene have been deposited in the GenBank database under GenBank Accession number; DQ421811.

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Authors and Affiliations

  1. Department of Molecular and Cell Biology, Boston University, Henry M. Goldman School of Dental Medicine, Boston, MA, USA

    Ahmad Almehmadi, Yoshio Ohyama, Ahmed Alamoudi, Dina Husein & Yoshiyuki Mochida

  2. Department of Periodontology and Oral Biology, Faculty of Dentistry, King Abdulaziz University, Jeddah, Saudi Arabia

    Ahmad Almehmadi

  3. Departoment of Maxillofacial Surgery, Tokyo Medical and Dental University Graduate School, Tokyo, Japan

    Yoshio Ohyama

  4. Division of Bio-Prosthodontics, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan

    Masaru Kaku

  5. Department of Oral Biology, Faculty of Dentistry, King Abdulaziz University, Jeddah, Saudi Arabia

    Ahmed Alamoudi

  6. Department of Oral Rehabilitation, Section of Fixed Prosthodontics, Fukuoka Dental College, Fukuoka, Japan

    Michitsuna Katafuchi

  7. Department of Biologic and Materials Sciences, School of Dentistry, University of Michigan, Ann Arbor, MI, USA

    Yuji Mishina

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  1. Ahmad Almehmadi
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Contributions

Conceived and designed the experiments: AA, YO, MK, YM. Performed the experiments: AA, YO, MK, AA, DH, MK, YM. Analyzed the data: AA, MK, YM, YM. Wrote the paper: AA, YO, YM, YM.

Corresponding author

Correspondence to Yoshiyuki Mochida.

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Conflict of interest

Ahmad Almehmadi, Yoshio Ohyama, Masaru Kaku, Ahmed Alamoudi, Dina Husein, Michitsuna Katafuchi, Yuji Mishina, Yoshiyuki Mochida declared that no competing financial interest.

Human and Animal Rights and Informed Consent

The use of animals and all animal procedures in this study were approved by the Institutional Animal Care and Use Committee (IACUC) of Boston University Medical campus (approved protocol number: AN-15053), and all efforts were made to minimize suffering animals. This study was carried out in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health.

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Almehmadi, A., Ohyama, Y., Kaku, M. et al. VWC2 Increases Bone Formation Through Inhibiting Activin Signaling. Calcif Tissue Int 103, 663–674 (2018). https://doi.org/10.1007/s00223-018-0462-9

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  • DOI: https://doi.org/10.1007/s00223-018-0462-9

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