Nupr1 deficiency downregulates HtrA1, enhances SMAD1 signaling, and suppresses age‐related bone loss in male mice

Nuclear protein 1 (NUPR1) is a stress‐induced protein activated by various stresses, such as inflammation and oxidative stress. We previously reported that Nupr1 deficiency increased bone volume by enhancing bone formation in 11‐week‐old mice. Analysis of differentially expressed genes between wild‐type (WT) and Nupr1‐knockout (Nupr1‐KO) osteocytes revealed that high temperature requirement A 1 (HTRA1), a serine protease implicated in osteogenesis and transforming growth factor‐β signaling was markedly downregulated in Nupr1‐KO osteocytes. Nupr1 deficiency also markedly reduced HtrA1 expression, but enhanced SMAD1 signaling in in vitro‐cultured primary osteoblasts. In contrast, Nupr1 overexpression enhanced HtrA1 expression in osteoblasts, suggesting that Nupr1 regulates HtrA1 expression, thereby suppressing osteoblastogenesis. Since HtrA1 is also involved in cellular senescence and age‐related diseases, we analyzed aging‐related bone loss in Nupr1‐KO mice. Significant spine trabecular bone loss was noted in WT male and female mice during 6−19 months of age, whereas aging‐related trabecular bone loss was attenuated, especially in Nupr1‐KO male mice. Moreover, cellular senescence‐related markers were upregulated in the osteocytes of 6−19‐month‐old WT male mice but markedly downregulated in the osteocytes of 19‐month‐old Nupr1‐KO male mice. Oxidative stress‐induced cellular senescence stimulated Nupr1 and HtrA1 expression in in vitro‐cultured primary osteoblasts, and Nupr1 overexpression enhanced p16ink4a expression in osteoblasts. Finally, NUPR1 expression in osteocytes isolated from the bones of patients with osteoarthritis was correlated with age. Collectively, these results indicate that Nupr1 regulates HtrA1‐mediated osteoblast differentiation and senescence. Our findings unveil a novel Nupr1/HtrA1 axis, which may play pivotal roles in bone formation and age‐related bone loss.