TMEM175 Plays a Crucial Role in Osteoblast Differentiation by Regulating Lysosomal Function and Autophagy

Bone provides structural support, enables movement, protects internal organs, regulates calcium and phosphorus levels, and contains bone marrow essential for hematopoiesis. Osteoblasts are specialized cells responsible for bone formation through the secretion of extracellular matrix components. Transmembrane protein 175 (TMEM175), which functions as an endosomal/lysosomal K⁺ channel and a lysosomal H⁺ channel, regulates lysosomal function and autophagy. Despite the recognized importance of lysosomes and autophagy in osteoblast differentiation, the specific role of TMEM175 in osteoblast differentiation has not been revealed. In this study, we investigated whether TMEM175 is associated with human bone mineral density (BMD) and fracture, and examined the role of TMEM175 in osteoblast differentiation. In analyses of single nucleotide polymorphisms (SNPs) of pore ion channel genes using the mouse2human database, a significant correlation between TMEM175 SNPs and human BMD and fracture was identified. TMEM175 expression levels were found to increase during osteoblast differentiation from bone chip-derived mesenchymal stem cells (BMSCs). Knockdown of TMEM175 in BMSCs suppressed osteoblast differentiation, as evidenced by decreased matrix mineralization and lower expression levels of osteoblast marker genes. Further analysis indicated that TMEM175 deficiency leads to lysosomal dysfunction and partially impairs autophagic clearance during osteoblast differentiation. Moreover, the TMEM175 inhibitor 4-aminopyridine (4-AP) decreased osteoblast differentiation of BMSCs. Taken together, this study reveals that TMEM175 plays an important role for osteoblast differentiation by regulating lysosomal function and autophagic clearance.