Bub1 suppresses inflammatory arthritis-associated bone loss in mice through inhibition of TNFα-mediated osteoclastogenesis.

Rheumatoid arthritis (RA) is an inflammatory autoimmune disease characterized by synovitis, bone and cartilage destruction, and increased fracture risk with bone loss. Although disease-modifying antirheumatic drugs have dramatically improved clinical outcomes, these therapies are not universally effective in all patients because of the heterogeneity of RA pathogenesis. Therefore, it is necessary to elucidate the molecular mechanisms underlying RA pathogenesis, including associated bone loss, in order to identify novel therapeutic targets. In this study, we found that Budding uninhibited by benzimidazoles 1 (BUB1) was highly expressed in RA patients' synovium and murine ankle tissue with arthritis. As CD45+CD11b+ myeloid cells are a Bub1 highly expressing population among synovial cells in mice, myeloid cell-specific Bub1 conditional knockout (Bub1ΔLysM) mice were generated. Bub1ΔLysM mice exhibited reduced femoral bone mineral density when compared with control (Ctrl) mice under K/BxN serum-transfer arthritis, with no significant differences in joint inflammation or bone erosion based on a semi-quantitative erosion score and histological analysis. Bone histomorphometry revealed that femoral bone mass of Bub1ΔLysM under arthritis was reduced by increased osteoclastic bone resorption. RNA-seq and subsequent Gene Set Enrichment Analysis demonstrated a significantly enriched nuclear factor-kappa B pathway among upregulated genes in receptor activator of nuclear factor kappa B ligand (RANKL)-stimulated bone marrow-derived macrophages (BMMs) obtained from Bub1ΔLysM mice. Indeed, osteoclastogenesis using BMMs derived from Bub1ΔLysM was enhanced by RANKL and tumor necrosis factor-α or RANKL and IL-1ß treatment compared with Ctrl. Finally, osteoclastogenesis was increased by Bub1 inhibitor BAY1816032 treatment in BMMs derived from wildtype mice. These data suggest that Bub1 expressed in macrophages plays a protective role against inflammatory arthritis-associated bone loss through inhibition of inflammation-mediated osteoclastogenesis.

Rheumatoid arthritis (RA) is a disease caused by an abnormal immune system, resulting in inflammation, swelling, and bone destruction in the joints, along with systemic bone loss. While new medications have dramatically improved treatment efficacy, these therapies are not universally effective for all patients. Therefore, we need to understand the regulatory mechanisms behind RA, including associated bone loss, to develop better therapies. In this study, we found that Budding uninhibited by benzimidazoles 1 (Bub1) was highly expressed in inflamed joints, especially in myeloid cells, which are a type of immune cells. To explore its role, we created myeloid cell­specific Bub1 conditional knockout (cKO) mice and induced arthritis to analyze its role during arthritis. The cKO mice exhibited lower bone mineral density when compared with control mice under inflammatory arthritis because of increased osteoclastic bone resorption, without significant differences in joint inflammation or bone erosion. Further investigation showed that Bub1 prevents excessive osteoclast differentiation induced by inflammation in bone marrow macrophages. These data suggest that Bub1 in macrophages protects against bone loss caused by inflammatory arthritis, offering potential insights for developing treatments that focus on bone health.

The effects of resistance training on bone mineral density and bone quality in type 2 diabetic rats.

Resistance training (RT) has been known to be effective in maintaining and improving bone strength, which is based on bone mineral density (BMD) and bone quality. However, it is not clear whether RT is effective in improving bone strength in patients with type-2 diabetes mellitus (T2DM), who have a high risk of fracture. Therefore, we tested the effects of a 6-week RT regimen using percutaneous electrical stimulation in T2DM model rats, male Otsuka Long-Evans Tokushima Fatty (OLETF), and its control, Long-Evans Tokushima Otsuka (LETO). After 6 weeks of RT, tibial BMD in RT legs was significantly higher than that in control (CON) legs in both groups. In diaphyseal cortical bone, bone area/tissue area, and cortical thickness was significantly increased in RT legs compared with CON legs in both groups. Cortical porosity was highly observed in OLETF compared with LETO, but RT improved cortical porosity in both groups. Interestingly, trabecular number, trabecular thickness and trabecular space as well as BMD and bone volume/tissue volume in proximal tibial metaphyseal trabecular bone were significantly improved in RT legs compared with CON legs in both groups. In contrast, connectivity density and structural model index were not affected by RT. These results indicate that the 6-week RT regimen effectively increased BMD and improved bone quality in T2DM model rats as well as control rats. Therefore, RT may have the potential to improve bone strength and reduce fracture risk, even in patients with T2DM.

The Effect of Ongoing Vitamin D and Low-Fat Milk Intake on Bone Metabolism in Female High-School Endurance Runners.

BACKGROUND: Vitamin D and calcium are essential nutrients for bone health. In addition, vitamin D suppresses inflammatory cytokines and increases bone resorption. Therefore, improvements in bone health by calcium and vitamin D supplementation have the potential to not only improve calcium metabolism but also suppress inflammation associated with exercise training. The purpose of this study was to determine whether ongoing vitamin D supplementation and low-fat milk intake by female high-school endurance runners would improve bone metabolism by suppressing inflammatory cytokines and the parathyroid hormone (PTH). METHODS: Twenty female high-school runners were assigned to a vitamin D supplement and low-fat milk intake group (MKD) or a control group (CON). Participants in the MKD group consumed a vitamin D supplement (1,000 IU/day) and low-fat milk (Ca 315 mg/day) for 6 months. Bone mineral density measurements, blood samples, and questionnaires (regarding menses and diet) were carried out. The UMIN Clinical Trials Registry number is UMIN000027854. RESULTS: The 25-hydroxyvitamin D (25(OH)D) concentration in MKD was sustained and PTH concentration was decreased regardless of the state of menses. The correlation coefficients of 25(OH)D or PTH concentrations and bone metabolism markers were analyzed by partial correlation coefficient via adjusting the model for frequency of menses. CTX and 25(OH)D concentration were significantly and inversely correlated at baseline (r = -0.61, P < 0.01), 3 months (r = -0.54, P = 0.02), and 6 months (r = -0.53, P = 0.02). CTX and PTH were significantly and positively correlated at 3 months (r = 0.63, P < 0.01) and 6 months (r = 0.52, P = 0.02). The bone alkaline phosphatase (BAP)/CTX ratio and 25(OH)D concentration were significantly and positively correlated at 3 months (r = 0.59, P = 0.01) and 6 months (r = 0.56, P = 0.01). The BAP/CTX ratio and PTH were significantly and inversely correlated at 3 months (r = -0.59, P = 0.01) and 6 months (r = -0.58, P < 0.01). CONCLUSIONS: This study suggested that vitamin D and low-fat milk supplementation improves bone metabolism by sustaining the 25(OH)D concentration and decreasing the PTH concentration in female high-school endurance runners regardless of the state of menses.