MiR‐664‐3p suppresses osteoblast differentiation and impairs bone formation via targeting Smad4 and Osterix

Osteoporosis is a metabolic disorder characterized by low bone mass and deteriorated microarchitecture, with an increased risk of fracture. Some miRNAs have been confirmed as potential modulators of osteoblast differentiation to maintain bone mass. Our miRNA sequencing results showed that miR‐664‐3p was significantly down‐regulated during the osteogenic differentiation of the preosteoblast MC3T3‐E1 cells. However, whether miR‐664‐3p has an impact on bone homeostasis remains unknown. In this study, we identified overexpression of miR‐664‐3p inhibited the osteoblast activity and matrix mineralization in vitro. Osteoblastic miR‐664‐3p transgenic mice exhibited reduced bone mass due to suppressed osteoblast function. Target prediction analysis and experimental validation confirmed Smad4 and Osterix (Osx) are the direct targets of miR‐664‐3p. Furthermore, specific inhibition of miR‐664‐3p by subperiosteal injection with miR‐664‐3p antagomir protected against ovariectomy‐induced bone loss. In addition, miR‐664‐3p expression was markedly higher in the serum from patients with osteoporosis compared to that from normal subjects. Taken together, this study revealed that miR‐664‐3p suppressed osteogenesis and bone formation via targeting Smad4 and Osx. It also highlights the potential of miR‐664‐3p as a novel diagnostic and therapeutic target for osteoporotic patients.