Qi Lingbin, Wang Jian, Chen Xian, Ding Yanhui, Ling Bin, Wang Wenjun, Xu Jun, Xue Zhigang
Tongji Hospital, School of Medicine, Tongji University, Shanghai 200092, China.
Shanghai East Hospital, Tongji University School of Medicine, Shanghai 200120, China.
Heliyon. 2022 Oct 18;8(11):e11148. doi: 10.1016/j.heliyon.2022.e11148. eCollection 2022 Nov.
Osteoarthritis (OA) is characterised by cartilage destruction; however, there are no specific drugs available for its treatment. Cartilage-derived stem/progenitor cells (CSPCs) are multipotent cells that play an essential role in cartilage renewal and may provide critical insights into the medical needs for OA treatment. However, alterations in cell function and fate of CSPCs during OA progression have seldom been analysed, especially at the single-cell level. Additionally, it has been reported that CSPCs can migrate to the cartilage injury area, although the mechanism of migration remains elusive. Thus, understanding the changing patterns of CSPCs in the pathological process of OA is important in the effort to develop stem cell therapy for OA. Here, we downloaded single-cell transcriptomic data of patients with OA from the Gene Expression Omnibus (GEO) database and performed unbiased clustering of the cells based on gene expression patterns using the Seurat package. Using common stem cell markers and chondrogenic transcription factors, we traced CSPCs throughout all stages of OA. We further explored the dynamics of CSPCs in OA progression and validated the single-cell RNA sequencing data using qPCR, immunofluorescence, and western blotting. Specifically, we primarily explored the heterogeneity of CSPCs at the single-cell level and found that it was closely associated with OA progression. Our results indicate significantly reduced chondrogenic differentiation capacity in CSPCs during the late stage of OA, while their proliferation capacity tended to increase. We also found that genes implicated in fibrosis, cell motility, and extracellular matrix remodelling were upregulated in CSPCs during the progression of OA. Our study revealed the dynamics of stem cells in OA progression and may inform the development of stem cell therapy for OA.
骨关节炎（OA）的特征是软骨破坏；然而，目前尚无用于治疗该病的特效药物。软骨来源的干/祖细胞（CSPCs）是多能细胞，在软骨更新中起重要作用，可能为OA治疗的医学需求提供关键见解。然而，很少有人分析OA进展过程中CSPCs的细胞功能和命运变化，尤其是在单细胞水平。此外，据报道CSPCs可以迁移到软骨损伤区域，但其迁移机制仍不清楚。因此，了解OA病理过程中CSPCs的变化模式对于开发OA的干细胞治疗方法很重要。在这里，我们从基因表达综合数据库（GEO）下载了OA患者的单细胞转录组数据，并使用Seurat软件包根据基因表达模式对细胞进行无偏聚类。我们使用常见的干细胞标记物和成软骨转录因子，追踪了OA各个阶段的CSPCs。我们进一步探讨了OA进展过程中CSPCs的动态变化，并通过qPCR、免疫荧光和蛋白质印迹法验证了单细胞RNA测序数据。具体而言，我们主要在单细胞水平上探索了CSPCs的异质性，发现其与OA进展密切相关。我们的结果表明，在OA晚期，CSPCs的成软骨分化能力显著降低，而其增殖能力则趋于增加。我们还发现，在OA进展过程中，与纤维化、细胞运动和细胞外基质重塑相关的基因在CSPCs中上调。我们的研究揭示了OA进展过程中干细胞的动态变化，可能为OA的干细胞治疗发展提供参考。
