Engineering osteoarthritic cartilage model through differentiating senescent human mesenchymal stem cells for testing disease-modifying drugs.
Engineering osteoarthritic cartilage model through differentiating senescent human mesenchymal stem cells for testing disease-modifying drugs.
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DOI：
10.1007/s11427-021-1933-7
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发表时间：
2022-02
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在从骨关节炎（OA）患者采集的软骨中已经观察到显著的细胞衰老。在这项研究中，我们的目标是开发一个衰老相关的OA样软骨模型，用于开发疾病修饰OA药物（DMOAD）。具体地，将人骨髓来源的间充质基质细胞（MSC）在体外扩增至第10代（P10-MSC）。在其衰老表型形成后，P10-MSC在软骨形成培养基中进行沉淀培养。来自qRT-PCR、组织学和免疫染色的结果表明，当与来自正常第4代（P4）-MSC的软骨相比时，在不使用其他OA诱导剂的情况下，来自P10-MSC的软骨显示衰老和OA样表型。有趣的是，在P4-MSC和P10-MSC衍生的软骨组织之间观察到的相同基因表达差异也在取自人样本的保存和受损OA软骨区域之间观察到，如RNA测序数据和其他分析方法所证明的。最后，通过测试几种潜在的DMOAD和senolytics来评估这种衰老引发的OA样软骨模型在药物开发中的效用。结果表明，预先存在的细胞衰老可以诱导软骨中OA样变化的产生。P4-和P10-MSC衍生的软骨模型也代表了用于预测潜在DMOAD对人类保存和受损软骨的功效和毒性的新平台。
Significant cellular senescence has been observed in cartilage harvested from patients with osteoarthritis (OA). In this study, we aim to develop a senescence-relevant OA-like cartilage model for developing disease-modifying OA drugs (DMOADs). Specifically, human bone marrow-derived mesenchymal stromal cells (MSCs) were expanded in vitro up to passage 10 (P10-MSCs). Following their senescent phenotype formation, P10-MSCs were subjected to pellet culture in chondrogenic medium. Results from qRT-PCR, histology and immunostaining indicated that cartilage generated from P10-MSCs displayed both senescent and OA-like phenotypes without using other OA-inducing agents, when compared to that from normal passage 4 (P4)-MSCs. Interestingly, the same gene expression differences observed between P4-MSCs and P10-MSC-derived cartilage tissues were also observed between the preserved and damaged OA cartilage regions taken from human samples, as demonstrated by RNA Sequencing data and other analysis methods. Lastly, the utility of this senescence-initiated OA-like cartilage model in drug development was assessed by testing several potential DMOADs and senolytics. The results suggest that pre-existing cellular senescence can induce the generation of OA-like changes in cartilage. The P4- and P10-MSCs derived cartilage models also represent a novel platform for predicting the efficacy and toxicity of potential DMOADs on both preserved and damaged cartilage in humans.