Department of Spine Surgery, Central Hospital of Dalian University of Technology, Dalian, China.
The First Affiliated Hospital of Dalian Medical University, Dalian, China.
Sci Rep. 2024 Sep 14;14(1):21489. doi: 10.1038/s41598-024-72474-0.
Age-related osteoporosis is a prevalent bone metabolic disorder distinguished by an aberration in the equilibrium between bone formation and resorption. The reduction in the stemness of Bone Marrow Mesenchymal Stem Cells (BMSCs) plays a pivotal role in the onset of this ailment. Comprehending the molecular pathways that govern BMSCs stemness is imperative for delineating the etiology of age-related osteoporosis and devising efficacious treatment modalities. The study utilized single-cell RNA sequencing and miRNA sequencing to investigate the cellular heterogeneity and stemness of BMSCs. Through dual-luciferase reporter assays and functional experiments, the regulatory effect of miR-183 on CTNNB1 (β-catenin) was confirmed. Overexpression and knockdown studies were conducted to explore the impact of miR-183 and β-catenin on stemness-related transcription factors Oct4, Nanog, and Sox2. Cell proliferation assays and osteogenic differentiation experiments were carried out to validate the influence of miR-183 and β-catenin on the stemness properties of BMSCs. Single-cell analysis revealed that β-catenin is highly expressed in both high stemness clusters and terminal differentiation clusters of BMSCs. Overexpression of β-catenin upregulated stemness transcription factors, while its suppression had the opposite effect, indicating a dual regulatory role of β-catenin in maintaining BMSCs stemness and promoting bone differentiation. Furthermore, the confluence of miRNA sequencing analyses and predictions from online databases revealed miR-183 as a potential modulator of BMSCs stemness and a novel upstream regulator of β-catenin. The overexpression of miR-183 effectively diminished the stemness characteristics of BMSCs by suppressing β-catenin, whereas the inhibition of miR-183 augmented stemness. These outcomes align with the observed alterations in the expression levels and functional assessments of transcription factors associated with stemness. This study provides evidence for the essential involvement of β-catenin in preserving the stemness of BMSCs, as well as elucidating the molecular mechanism through which miR-183 selectively targets β-catenin to modulate stemness. These results underscore the potential of miR-183 and β-catenin as molecular targets for augmenting the stemness of BMSCs. This strategy is anticipated to facilitate the restoration of bone microarchitecture and facilitate bone tissue regeneration by addressing potential cellular dysfunctions, thereby presenting novel targets and perspectives for the management of age-related osteoporosis.
与年龄相关的骨质疏松症是一种常见的骨骼代谢紊乱疾病，其特征是骨形成和吸收之间的平衡发生了偏差。骨髓间充质干细胞（BMSCs）干性的降低在这种疾病的发生中起着关键作用。理解控制 BMSCs 干性的分子途径对于阐明与年龄相关的骨质疏松症的病因和设计有效的治疗方法至关重要。本研究利用单细胞 RNA 测序和 miRNA 测序技术研究了 BMSCs 的细胞异质性和干性。通过双荧光素酶报告基因检测和功能实验，证实了 miR-183 对 CTNNB1（β-catenin）的调控作用。通过过表达和敲低研究，探讨了 miR-183 和β-catenin 对干性相关转录因子 Oct4、Nanog 和 Sox2 的影响。细胞增殖实验和成骨分化实验验证了 miR-183 和β-catenin 对 BMSCs 干性特性的影响。单细胞分析表明，β-catenin 在 BMSCs 的高干性簇和终末分化簇中均高度表达。β-catenin 的过表达上调了干性转录因子，而其抑制则产生相反的效果，表明β-catenin 在维持 BMSCs 干性和促进骨分化方面具有双重调控作用。此外，miRNA 测序分析与在线数据库预测结果的结合揭示了 miR-183 作为 BMSCs 干性的潜在调节剂和β-catenin 的新型上游调节剂。miR-183 的过表达通过抑制β-catenin 有效降低了 BMSCs 的干性特征，而 miR-183 的抑制则增强了干性。这些结果与与干性相关的转录因子的表达水平和功能评估的变化一致。本研究为β-catenin 在维持 BMSCs 干性中的重要作用提供了证据，并阐明了 miR-183 通过选择性靶向β-catenin 来调节干性的分子机制。这些结果强调了 miR-183 和β-catenin 作为增强 BMSCs 干性的分子靶标的潜力。该策略有望通过解决潜在的细胞功能障碍来促进骨微结构的恢复和促进骨组织再生，为与年龄相关的骨质疏松症的治疗提供新的靶点和思路。
