DZNep promotes mouse bone defect healing via enhancing both osteogenesis and osteoclastogenesis

Stem Cell Research & Therapy

Shenggui Xu¹  Yuwei Liang²  Shuai Han³  Wenxin He⁴  Kewei Rong⁴  An Qin⁴  Xiankun Cao⁴  Xiao Yang⁴  Hui Ma⁴  Zhiqian Chen⁴  Jie Zhao⁴  Yifan Zhou⁴ 

[1] Department of Orthopaedics, Mindong Hospital Affiliated to Fujian Medical University, 355000, Fuan, Fujian Province, People’s Republic of China;Department of Orthopedics, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, 510120, Guangzhou, People’s Republic of China;Guangxi Key Laboratory of Regenerative Medicine, Guangxi Collaborative Innovation Center for Biomedicine, GuangxiASEAN Collaborative Innovation Center for Major Disease Prevention and Treatment, Guangxi Medical University, 530021, Nanning, Guangxi, People’s Republic of China;Shanghai Key Laboratory of Orthopedic Implants, Department of Orthopaedics Surgery, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, No. 639, Zhizaoju Road, 200011, Shanghai, People’s Republic of China;

关键词: DZNep;    Osteoclast;    Osteoblast;    Bone defect;    EZH2-H3K27me3-Wnt signaling pathway;    EZH2-H3K27me3-Foxc1-NF-κB signaling pathway;   

DOI  :  10.1186/s13287-021-02670-6

来源: Springer

【 摘 要 】

BackgroundEnhancer of zeste homolog 2 (EZH2) is a novel oncogene that can specifically trimethylate the histone H3 lysine 27 (H3K27me3) to transcriptionally inhibit the expression of downstream tumor-suppressing genes. As a small molecular inhibitor of EZH2, 3-Deazaneplanocin (DZNep) has been widely studied due to the role of tumor suppression. With the roles of epigenetic regulation of bone cells emerged in past decades, the property and molecular mechanism of DZNep on enhancing osteogenesis had been reported and attracted a great deal of attention recently. This study aims to elucidate the role of DZNep on EZH2-H3K27me3 axis and downstream factors during both osteoclasts and osteoblasts formation and the therapeutic possibility of DZNep on bone defect healing.MethodsBone marrow-derived macrophages (BMMs) cells were cultured, and their responsiveness to DZNep was evaluated by cell counting kit-8, TRAP staining assay, bone resorption assay, podosome actin belt. Bone marrow-derived mesenchymal stem cells (BMSC) were cultured and their responsiveness to DZNep was evaluated by cell counting kit-8, ALP and AR staining assay. The expression of nuclear factor-κB (NF-κB), mitogen-activated protein kinase (MAPK), Wnt signaling pathway was determined by qPCR and western blotting. Mouse bone defect models were created, rescued by DZNep injection, and the effectiveness was evaluated by X-ray and micro-CT and histological staining.ResultsConsistent with the previous study that DZNep enhances osteogenesis via Wnt family member 1(Wnt1), Wnt6, and Wnt10a, our results showed that DZNep also promotes osteoblasts differentiation and mineralization through the EZH2-H3K27me3-Wnt4 axis. Furthermore, we identified that DZNep promoted the receptor activator of nuclear factor-κB (NF-κB) ligand (RANKL)-induced osteoclast formation via facilitating the phosphorylation of IKKα/β, IκB, and subsequently NF-κB nuclear translocation, which credit to the EZH2-H3K27me3-Foxc1 axis. More importantly, the enhanced osteogenesis and osteoclastogenesis result in accelerated mice bone defect healing in vivo.ConclusionDZNep targeting EZH2-H3K27me3 axis facilitated the healing of mice bone defect via simultaneously enhancing osteoclastic bone resorption and promoting osteoblastic bone formation.

【 授权许可】

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