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New Function of RUNX2 in Regulating Osteoclast Differentiation via the AKT/NFATc1/CTSK Axis

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Abstract

Cleidocranial dysplasia is an autosomal dominant skeletal disorder resulting from RUNX2 mutations. The influence of RUNX2 mutations on osteoclastogenesis and bone resorption have not been reported. To investigate the role of RUNX2 in osteoclast, RUNX2 expression in macrophages (RAW 264.7 cells) was detected. Stable RAW 264.7 cell lines expressing wild-type RUNX2 or mutated RUNX2 (c.514delT, p.172 fs) were established, and their functions in osteoclasts were investigated. Wild-type RUNX2 promoted osteoclast differentiation, formation of F-actin ring, and bone resorption, while mutant RUNX2 attenuated the positive differentiation effect. Wild-type RUNX2 increased the expression and activity of mTORC2. Subsequently, mTORC2 specifically promoted phosphorylation of AKT at the serine 473 residue. Activated AKT improved the nuclear translocation of NFATc1 and increased the expression of downstream genes, including CTSK. Inhibition of AKT phosphorylation abrogated the osteoclast formation of wild-type macrophages, whereas constitutively activated AKT rescued the osteoclast formation of mutant macrophages. The present study suggested that RUNX2 promotes osteoclastogenesis and bone resorption through the AKT/NFATc1/CTSK axis. Mutant RUNX2 lost the function of regulating osteoclast differentiation and bone remodeling, resulting in the defective formation of the tooth eruption pathway and impaction of permanent teeth in cleidocranial dysplasia. This study, for the first time, verifies the effect of RUNX2 on osteoclast differentiation and bone resorption and provides new insight for the explanation of cleidocranial dysplasia.

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Acknowledgments

This work was supported by the grants of National Natural Science Foundation of China (grant number 81771053, 81772873 and 81970920). The authors are grateful to all the participants in this study.

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Authors and Affiliations

  1. Department of Preventive Dentistry, Peking University School and Hospital of Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Laboratory for Digital and Material Technology of Stomatology & Beijing Key Laboratory of Digital Stomatology, Beijing, People’s Republic of China

    Yuejiao Xin, Yang Liu, Dandan Liu, Jie Li, Chenying Zhang & Shuguo Zheng

  2. Central Laboratory, Department of Oral and Maxillofacial Surgery, Peking University School and Hospital of Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Laboratory for Digital and Material Technology of Stomatology & Beijing Key Laboratory of Digital Stomatology, Beijing, People’s Republic of China

    Yixiang Wang

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  1. Yuejiao Xin
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Contributions

Conceptualization: YX, YW and SZ; Methodology: YX, DL, JL and CZ; Data Curation: YX and YL; Writing—Original Draft Preparation: YX; Writing—Review & Editing: YW and SZ.

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Correspondence to Yixiang Wang or Shuguo Zheng.

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Yuejiao Xin, Yang Liu, Dandan Liu, Jie Li, Chenying Zhang, Yixiang Wang, and Shuguo Zheng declare that they have no conflict of interest.

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This study was approved by the Ethical Committee of Peking University School and the Hospital of Stomatology (Approval No. PKUSSIRB-2012004) and conducted strictly in accordance with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. All participants signed an informed consent prior to study.

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Xin, Y., Liu, Y., Liu, D. et al. New Function of RUNX2 in Regulating Osteoclast Differentiation via the AKT/NFATc1/CTSK Axis. Calcif Tissue Int 106, 553–566 (2020). https://doi.org/10.1007/s00223-020-00666-7

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