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Hedgehog signaling regulates bone homeostasis through orchestrating osteoclast differentiation and osteoclast–osteoblast coupling

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Abstract

Imbalance of bone homeostasis induces bone degenerative diseases such as osteoporosis. Hedgehog (Hh) signaling plays critical roles in regulating the development of limb and joint. However, its unique role in bone homeostasis remained largely unknown. Here, we found that canonical Hh signaling pathway was gradually augmented during osteoclast differentiation. Genetic inactivation of Hh signaling in osteoclasts, using Ctsk-Cre;Smof/f conditional knockout mice, disrupted both osteoclast formation and subsequent osteoclast–osteoblast coupling. Concordantly, either Hh signaling inhibitors or Smo/Gli2 knockdown stunted in vitro osteoclast formation. Mechanistically, Hh signaling positively regulated osteoclast differentiation via transactivation of Traf6 and stabilization of TRAF6 protein. Then, we identified connective tissue growth factor (CTGF) as an Hh-regulatory bone formation-stimulating factor derived from osteoclasts, whose loss played a causative role in osteopenia seen in CKO mice. In line with this, recombinant CTGF exerted mitigating effects against ovariectomy induced bone loss, supporting a potential extension of local rCTGF treatment to osteoporotic diseases. Collectively, our findings firstly demonstrate that Hh signaling, which dictates osteoclast differentiation and osteoclast–osteoblast coupling by regulating TRAF6 and CTGF, is crucial for maintaining bone homeostasis, shedding mechanistic and therapeutic insights into the realm of osteoporosis.

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Data availability

The data that support the findings of this study are available from the corresponding author LY upon reasonable request.

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Acknowledgements

We thank Prof. Baojie Li for providing Smoflox/flox mice. We also thank Prof. Minghao Zheng for a kind gift of bovine bone slices.

Funding

This research was financially supported by the National Natural Science Foundation of China (81772377, 81871743, 81902202, 82130070), Innovation Team Projects—Innovation Capability Support Program of Shaanxi Province (2020TD-036), and Clinical Medical Research Center Projects—Innovation Capability Support Program of Shaanxi Province (2020LCZX-03). Key Research and Development Program of Shaanxi (2021SF-024).

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Author notes

  1. Weiguang Lu, Chao Zheng and Hongyang Zhang contributed equally.

Authors and Affiliations

  1. Department of Orthopedics, Xijing Hospital, Fourth Military Medical University, Xi’an, China

    Weiguang Lu, Chao Zheng, Hongyang Zhang, Pengzhen Cheng, Sheng Miao, Huanbo Wang, Jing Fan, Yaqian Hu, He Liu, Zhuojing Luo & Liu Yang

  2. Pediatric Orthopaedic Hospital, Honghui Hospital, Xi’an Jiaotong University, Xi’an, China

    Pengzhen Cheng, Xue Hao & Qiang Jie

  3. Research Center for Skeletal Developmental Deformity and Injury Repair, College of Life Science and Medicine, Northwest University, Xi’an, China

    Pengzhen Cheng, Liyuan Jia & Qiang Jie

  4. Clinical Research Center for Pediatric Skeletal Deformity and Injury of Shaanxi Province, Xi’an, China

    Qiang Jie

  5. Institute of Medical Research, Northwestern Polytechnical University, Xi’an, China

    Ting He

  6. School of Biomedical Sciences, University of Western Australia, Perth, WA, 6009, Australia

    Jiake Xu

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  1. Weiguang Lu
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Contributions

WL, QJ and LY designed the study and wrote and prepared the manuscript and figures. HZ and HW performed experiments on CKO mice. WL and CZ assisted experiments including western blot and qPCR in vitro. PC and CZ provided support about histomorphology and microCT analysis. SM and TH contributed to the operation and treatment. JF and HL performed mouse feeding and genotyping. YH and XH provided statistical analysis. LJ drew a schematic picture. ZL and JX revised this manuscript.

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Correspondence to Qiang Jie or Liu Yang.

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Lu, W., Zheng, C., Zhang, H. et al. Hedgehog signaling regulates bone homeostasis through orchestrating osteoclast differentiation and osteoclast–osteoblast coupling. Cell. Mol. Life Sci. 80, 171 (2023). https://doi.org/10.1007/s00018-023-04821-9

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