Genetic disruption of Ano5 leads to impaired osteoclastogenesis for gnathodiaphyseal dysplasia

• Published in: Oral diseases Vol. 30; no. 3; pp. 1403 - 1415
• Main Authors: Liu, Xiu, Wang, Xiaoyu, Ma, Xinrong, Li, Hongyu, Miao, Congcong, Tian, Zhenchuan, Hu, Ying
• Format: Journal Article
• Language: English
• Published: Denmark Wiley Subscription Services, Inc 01.04.2024
• Subjects: Actin; Animals; Ano5; Anoctamins - genetics; Bone resorption; Bone Resorption - genetics; Bone Resorption - pathology; Bone turnover; Cathepsin K - genetics; Cell Differentiation - genetics; Dysplasia; Gelatinase B; Gene expression; genetic animal model; Genetic disorders; gnathodiaphyseal dysplasia; Homeostasis; Immunohistochemistry; Macrophages; Membrane Proteins - genetics; Membrane Proteins - metabolism; Mice; Mice, Knockout; Multiple sclerosis; Nerve Tissue Proteins; NFATC Transcription Factors - genetics; NFATC Transcription Factors - metabolism; Osteoblastogenesis; osteoclast; Osteoclastogenesis; Osteoclasts; Osteoclasts - metabolism; Osteogenesis - genetics; Osteogenesis Imperfecta - genetics; Osteogenesis Imperfecta - pathology; Phalloidin; Phenotypes; Polymerase chain reaction; Proto-Oncogene Proteins c-fos - genetics; Proto-Oncogene Proteins c-fos - metabolism; RANK Ligand - genetics; RANK Ligand - metabolism; Sclerosis; Signal Transduction; signalling pathway; Tartrate-Resistant Acid Phosphatase - genetics; Tartrate-Resistant Acid Phosphatase - metabolism; genetic animal model; signalling pathway; Ano5; osteoclast; gnathodiaphyseal dysplasia
• ISSN: 1354-523X; 1601-0825; 1601-0825
• DOI: 10.1111/odi.14562

Objectives Gnathodiaphyseal dysplasia (GDD; OMIM#166260) is a rare skeletal genetic disorder characterized by sclerosis of tubular bones and cemento‐osseous lesions in mandibles. TMEM16E/ANO5 gene mutations have been identified in patients with GDD. Here, Ano5 knockout (Ano5−/−) mice with enhanced osteoblastogenesis were used to investigate whether Ano5 disruption affects osteoclastogenesis. Subjects and Methods The maturation of osteoclasts, formation of F‐actin ring and bone resorption were detected by immunohistochemistry, TRAP, phalloidin staining and Coming Osteo assays. The expression of osteoclast‐related factors was measured by qRT‐PCR. Early signaling pathways were verified by western blot. Results Ano5−/− mice exhibited inhibitory formation of multinucleated osteoclasts with a reduction of TRAP activity. The expression of Nfatc1, c‐Fos, Trap, Ctsk, Mmp9, Rank and Dc‐stamp was significantly decreased in bone tissues and bone marrow‐derived macrophages (BMMs) of Ano5−/− mice. Ano5−/− osteoclasts manifested disrupted actin ring and less mineral resorption. RANKL‐induced early signaling pathways were suppressed in Ano5−/− osteoclasts and Ano5 knockdown RAW264.7 cells. Moreover, the inhibitory effects of NF‐κB signalling pathway on osteoclastogenesis were partially attenuated with NF‐κB signalling activator. Conclusions Ano5 deficiency impairs osteoclastogenesis, which leads to enhanced osteogenic phenotypes mediated by bone homeostasis dysregulation.