Wang Shan, Wang Ziming, Su Hongjun, Chen Fenglei, Ma Mengjun, Yu Wenhui, Ye Guiwen, Cen Shuizhong, Mi Rujia, Wu Xiaohua, Deng Wen, Feng Pei, Zeng Chenying, Shen Huiyong, Wu Yanfeng
Center for Biotherapy, Eighth Affiliated Hospital of Sun Yat-sen University, Shenzhen 518033, P.R. China.
Department of Orthopedics, Eighth Affiliated Hospital of Sun Yat-sen University, Shenzhen 518033, P.R. China.
Mol Ther Nucleic Acids. 2021 Aug 19;26:557-574. doi: 10.1016/j.omtn.2021.08.013. eCollection 2021 Dec 3.
Expansion prior to mesenchymal stem cells (MSCs) application is a necessary process. Functional and genomic stability has a crucial role in stem-cell-based therapies. However, the exact expression and co-expressed profiles of coding and non-coding RNAs in human bone marrow (BM)-MSCs aging are still lacking. In the present studies, the change of morphology, immunophenotype, and capacity of proliferation, differentiation, and immunoregulation of MSCs at passage (P) 4, P6, P8, P10, and P12 were investigated. RNA sequencing identified that 439 mRNAs, 65 long noncoding RNAs (lncRNAs), 59 microRNAs (miRNAs), and 229 circular RNAs (circRNAs) were differentially expressed (DE) in P12 compared with P4, with a similar trend in P6. Gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and gene set enrichment analysis (GSEA) identified several significant biological processes and pathways, including binding, ossification, and Wnt and PPAR signaling pathways. Interaction and co-expression/localization analyses were performed for DE mRNAs and lncRNAs, and several key lncRNAs, circRNAs, and important pathways like autophagy and mitophagy were identified in the competing endogenous RNA (ceRNA) network. Some key RNAs found in the bioinformatics analysis were validated. Our studies indicate that replicative senescence of MSCs is a continuous process, including widespread alterations in biological characteristics and global gene expression patterns that need to be considered before therapeutic applications of MSCs.
间充质干细胞（MSCs）应用前的扩增是一个必要过程。功能和基因组稳定性在基于干细胞的治疗中起着关键作用。然而，人类骨髓（BM）-MSCs衰老过程中编码和非编码RNA的确切表达及共表达谱仍不清楚。在本研究中，对第4代、第6代、第8代、第10代和第12代MSCs的形态、免疫表型以及增殖、分化和免疫调节能力的变化进行了研究。RNA测序确定，与第4代相比，第12代中有439个mRNA、65个长链非编码RNA（lncRNA）、59个微小RNA（miRNA）和229个环状RNA（circRNA）差异表达（DE），第6代也有类似趋势。基因本体论（GO）、京都基因与基因组百科全书（KEGG）和基因集富集分析（GSEA）确定了几个重要的生物学过程和途径，包括结合、骨化以及Wnt和PPAR信号通路。对DE mRNA和lncRNA进行了相互作用和共表达/定位分析，并在竞争性内源RNA（ceRNA）网络中鉴定了几个关键的lncRNA、circRNA以及自噬和线粒体自噬等重要途径。对生物信息学分析中发现的一些关键RNA进行了验证。我们的研究表明，MSCs的复制性衰老为一个连续过程，包括生物学特性和整体基因表达模式的广泛改变，这些在MSCs治疗应用前均需加以考虑。
