Fukui Tatsuyuki, Terashima Asuka, Omata Yasunori, Chijimatsu Ryota, Okamoto Kazuo, Tsukasaki Masayuki, Fukuda Yukiko, Hayata Tadayoshi, Saitoh Akiyoshi, Toda Etsuko, Takayanagi Hiroshi, Tanaka Sakae, Terashima Yuya, Saito Taku
Sensory and Motor System Medicine, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8655, Japan.
Bone and Cartilage Regenerative Medicine, The University of Tokyo Hospital, Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8655, Japan.
J Bone Miner Metab. 2025 Mar;43(2):61-73. doi: 10.1007/s00774-024-01555-x. Epub 2024 Oct 7.
Disulfiram (DSF), known as an anti-alcoholism drug, has been reported to suppress osteoclast differentiation in vitro; however, it remains uncertain whether DSF is effective in preventing osteoclastogenesis in vivo. This study aimed to investigate the effect of DSF administration in osteoporotic mice and its contribution to osteoclastogenesis in vivo.
The bone phenotype of ovariectomized mice, both treated and untreated with DSF, was examined using microcomputed tomography analysis. Osteoclastic and osteoblastic parameters were assessed through bone morphometric analysis. The direct effect of DSF on osteoblastogenesis in vitro was evaluated via a primary osteoblast culture experiment. The expression of genes related to DSF targets (Nup85, Ccr2, and Ccr5) in osteoclast-lineage cells was examined using scRNA-seq analysis and flow cytometry analysis using the bone marrow cells from ovariectomized mice. The impact of DSF on osteoclast-lineage cells was assessed using primary cultures of osteoclasts.
DSF administration ameliorated ovariectomy-induced bone loss and mitigated the increase of osteoclasts without affecting osteoblastogenesis. The scRNA-seq data revealed that osteoclast precursor cells expressed Nup85, Ccr2, and Ccr5. CCR2 and CCR5-positive cells in osteoclast precursor cells within bone marrow increased following ovariectomy, and this increase was canceled by DSF administration. Finally, we found that DSF had a significant inhibitory effect on osteoclastogenesis in the early stage by suppressing Tnfrsf11a expression.
This study demonstrates that DSF could be a candidate for osteoporosis therapies because it suppresses osteoclastogenesis from an early stage in vivo.
双硫仑（DSF）作为一种戒酒药物，已被报道在体外可抑制破骨细胞分化；然而，DSF在体内预防破骨细胞生成是否有效仍不确定。本研究旨在探讨DSF给药对骨质疏松小鼠的影响及其在体内对破骨细胞生成的作用。
使用微型计算机断层扫描分析检查经DSF治疗和未治疗的去卵巢小鼠的骨表型。通过骨形态计量分析评估破骨细胞和成骨细胞参数。通过原代成骨细胞培养实验评估DSF对体外成骨细胞生成的直接作用。使用单细胞RNA测序（scRNA-seq）分析和流式细胞术分析，利用去卵巢小鼠的骨髓细胞检测破骨细胞谱系细胞中与DSF靶点（Nup85、Ccr2和Ccr5）相关的基因表达。使用破骨细胞原代培养评估DSF对破骨细胞谱系细胞的影响。
DSF给药改善了去卵巢诱导的骨质流失，并减轻了破骨细胞的增加，而不影响成骨细胞生成。scRNA-seq数据显示破骨细胞前体细胞表达Nup85、Ccr2和Ccr5。去卵巢后骨髓中破骨细胞前体细胞中CCR2和CCR5阳性细胞增加，而DSF给药可消除这种增加。最后，我们发现DSF通过抑制Tnfrsf11a表达在早期对破骨细胞生成具有显著抑制作用。
本研究表明DSF可能是骨质疏松症治疗的候选药物，因为它在体内从早期阶段就抑制破骨细胞生成。
