Genetic disruption of Ano5 leads to impaired osteoclastogenesis for gnathodiaphyseal dysplasia

Abstract

AbstractObjectivesGnathodiaphyseal 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 MethodsThe 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.ResultsAno5−/− 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.ConclusionsAno5 deficiency impairs osteoclastogenesis, which leads to enhanced osteogenic phenotypes mediated by bone homeostasis dysregulation.