A transcriptomic analysis of serial-cultured, tonsil-derived mesenchymal stem cells reveals decreased integrin α3 protein as a potential biomarker of senescent cells
A transcriptomic analysis of serial-cultured, tonsil-derived mesenchymal stem cells reveals decreased integrin α3 protein as a potential biomarker of senescent cells
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DOI：
10.1186/s13287-020-01860-y
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发表时间：
2020-08-17
影响因子：
7.5
通讯作者：
Park, Yoon Shin
中科院分区：
文献类型：
--
作者：
Choi, Da Hyeon;Oh, Se-Young;Park, Yoon Shin
背景资料：间充质干细胞（MSC）已被广泛用于干细胞治疗，并且通常需要连续传代干细胞以获得足够的细胞数量用于再生医学中的实际应用。长期连续细胞扩增可潜在地诱导复制性衰老，这导致干细胞功能和干细胞性的进行性下降，失去多能性特征。为了提高干细胞治疗的疗效，识别衰老细胞的特异性生物标志物将是重要的。方法：将20-25代的扁桃体来源的间充质干细胞（TMSCs）指定为培养老化的TMSCs，并将其中胚层分化潜能以及衰老和干细胞标志物与最多传代8次的对照TMSCs（指定为年轻）进行比较。全基因组分析被用来确定新的调节因子，区分培养年龄和控制TMSC。使用Western blot analysis.Results：培养老化的TMSCs显示较长的倍增时间相比，对照TMSCs，并有较高的表达衰老相关（SA）-β-半乳糖染色，但较低的表达的干性蛋白标志物，包括Nanog，Oct 4，和Sox 2的脂肪，成骨和软骨分化潜能降低。微阵列分析确定了培养老化和对照TMSC之间总共18，614个差异表达基因。利用KEGG（京都基因和基因组百科全书）途径分析将差异表达基因分为细胞组分（CC）、功能组分（FC）和生物过程（BP）三类。该分析显示，与CC和BP相关的那些基因在培养老化的TMSC和对照TMSC之间显示出最显著的差异。与细胞外基质-受体相互作用相关的基因也显示出显著差异（p < 0.001）。我们还发现，培养老化TMSCs的整合素α 3（ITGA 3）和磷酸化AKT蛋白（p-AKT-Ser（473））的表达下降相比，控制TMSCs.Conclusions：我们的数据表明，ECM受体信号的激活，特别是参与整合素家族介导的细胞内细胞生存信号分子AKT的激活，可以调节干细胞衰老TMSCs。在这些鉴定的因子中，发现ITGA 3是衰老TMSC的代表性生物标志物。在本研究中排除具有衰老TMSC标志物的TMSC可能潜在地增加TMSC在临床应用中的治疗功效。
Background: Mesenchymal stem cells (MSCs) have been widely used for stem cell therapy, and serial passage of stem cells is often required to obtain sufficient cell numbers for practical applications in regenerative medicine. A long-term serial cell expansion can potentially induce replicative senescence, which leads to a progressive decline in stem cell function and stemness, losing multipotent characteristics. To improve the therapeutic efficiency of stem cell therapy, it would be important to identify specific biomarkers for senescent cells.Methods: Tonsil-derived mesenchymal stem cells (TMSCs) with 20-25 passages were designated as culture-aged TMSCs, and their mesodermal differentiation potentials as well as markers of senescence and stemness were compared with the control TMSCs passaged up to 8 times at the most (designated as young). A whole-genome analysis was used to identify novel regulatory factors that distinguish between the culture-aged and control TMSCs. The identified markers of replicative senescence were validated using Western blot analyses.Results: The culture-aged TMSCs showed longer doubling time compared to control TMSCs and had higher expression of senescence-associated (SA)-beta-gal staining but lower expression of the stemness protein markers, including Nanog, Oct4, and Sox2 with decreased adipogenic, osteogenic, and chondrogenic differentiation potentials. Microarray analyses identified a total of 18,614 differentially expressed genes between the culture-aged and control TMSCs. The differentially expressed genes were classified into the Gene Ontology categories of cellular component (CC), functional component (FC), and biological process (BP) using KEGG (Kyoto encyclopedia of genes and genomes) pathway analysis. This analysis revealed that those genes associated with CC and BP showed the most significant difference between the culture-aged and control TMSCs. The genes related to extracellular matrix-receptor interactions were also shown to be significantly different (p < 0.001). We also found that culture-aged TMSCs had decreased expressions of integrin alpha 3 (ITGA3) and phosphorylated AKT protein (p-AKT-Ser(473)) compared to the control TMSCs.Conclusions: Our data suggest that activation of ECM-receptor signaling, specifically involved with integrin family-mediated activation of the intracellular cell survival-signaling molecule AKT, can regulate stem cell senescence in TMSCs. Among these identified factors, ITGA3 was found to be a representative biomarker of the senescent TMSCs. Exclusion of the TMSCs with the senescent TMSC markers in this study could potentially increase the therapeutic efficacy of TMSCs in clinical applications.