Cen Shuizhong, Cai Mingxi, Wang Yihan, Lu Xiuyi, Chen Zhipeng, Chen Haobo, Fang Yingdong, Wu Changping, Qiu Sujun, Liu Zhenhua
Department of Spine Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou, China.
Department of Dermatology, The Fourth Affiliated Hospital of Guangzhou Medical University, Guangzhou, China.
Front Genet. 2022 Oct 3;13:991875. doi: 10.3389/fgene.2022.991875. eCollection 2022.
We have already demonstrated that mesenchymal stem cells from patients with ankylosing spondylitis (ASMSCs) exhibited greater adipogenic differentiation potential than those from healthy donors (HDMSCs). Here, we further investigated the expression profile of long noncoding RNA (lncRNA) and mRNA, aiming to explore the underlying mechanism of abnormal adipogenic differentiation in ASMSCs. HDMSCs and ASMSCs were separately isolated and induced with adipogenic differentiation medium for 10 days. Thereafter, lncRNAs and mRNAs that were differentially expressed (DE) between HDMSCs and ASMSCs were identified high-throughput sequencing and confirmed by quantitative real-time PCR (qRT-PCR) assays. Then, the DE genes were annotated and enriched by GO analysis. In addition, protein interaction network was constructed to evaluate the interactions between DE mRNAs and to find hub nodes and study cliques. Besides, co-expression network analysis was carried out to assess the co-expressions between DE mRNA and DE lncRNAs, and competing endogenous RNA (ceRNA) network analysis were conducted to predict the relationships among lncRNAs, mRNAs and miRNAs. The signaling pathways based on the DE genes and the predicted DE genes were enriched by KEGG analysis. A total of 263 DE lncRNAs and 1376 DE mRNAs were found during adipogenesis in ASMSCs. qRT-PCR indicated that the expression of the top 20 mRNAs and the top 10 lncRNAs was consistent with the high-throughput sequencing data. Several lncRNAs (NR_125386.1, NR_046473.1 and NR_038937.1) and their target genes (SPN and OR1AIP2), together with the significantly co-expressed pairs of DE lncRNAs and DE mRNAs (SLC38A5-ENST00000429588.1, TMEM61-ENST00000400755.3 and C5orf46-ENST00000512300.1), were closely related to the enhanced adipogenesis of ASMSCs by modulating the PPAR signaling pathway. Our study analyzed the expression profiles of DE lncRNAs and DE mRNAs during adipogenesis in ASMSCs and HDMSCs. Several DE lncRNAs, DE mRNAs and signaling pathways that probably participate in the aberrant adipogenesis of ASMSCs were selected for future study. These results will likely provide potential targets for our intervention on fat metaplasia and subsequent new bone formation in patients with AS in the future.
我们已经证明，强直性脊柱炎患者的间充质干细胞（ASMSCs）比健康供体的间充质干细胞（HDMSCs）表现出更大的成脂分化潜能。在此，我们进一步研究长链非编码RNA（lncRNA）和mRNA的表达谱，旨在探索ASMSCs中异常成脂分化的潜在机制。分别分离HDMSCs和ASMSCs，并用成脂分化培养基诱导10天。此后，通过高通量测序鉴定HDMSCs和ASMSCs之间差异表达（DE）的lncRNAs和mRNAs，并通过定量实时PCR（qRT-PCR）分析进行验证。然后，通过GO分析对DE基因进行注释和富集。此外，构建蛋白质相互作用网络以评估DE mRNAs之间的相互作用，并找到枢纽节点和研究团块。此外，进行共表达网络分析以评估DE mRNA和DE lncRNAs之间的共表达，并进行竞争性内源RNA（ceRNA）网络分析以预测lncRNAs、mRNAs和miRNAs之间的关系。基于DE基因和预测的DE基因的信号通路通过KEGG分析进行富集。在ASMSCs成脂过程中总共发现了263个DE lncRNAs和1376个DE mRNAs。qRT-PCR表明前20个mRNAs和前10个lncRNAs的表达与高通量测序数据一致。几个lncRNAs（NR_125386.1、NR_046473.1和NR_038937.1）及其靶基因（SPN和OR1AIP2），以及DE lncRNAs和DE mRNAs的显著共表达对（SLC38A5-ENST00000429588.1、TMEM61-ENST00000400755.3和C5orf46-ENST00000512300.1），通过调节PPAR信号通路与ASMSCs增强的成脂密切相关。我们的研究分析了ASMSCs和HDMSCs成脂过程中DE lncRNAs和DE mRNAs的表达谱。选择了几个可能参与ASMSCs异常成脂的DE lncRNAs、DE mRNAs和信号通路以供未来研究。这些结果可能为我们未来干预AS患者的脂肪化生和随后的新骨形成提供潜在靶点。
