Govarthanan Kavitha, Gupta Piyush Kumar, Patra Bamadeb, Ramasamy Deepa, E Binita Zipporah, Sharma Vineeta, Yadav Rajesh, Kumar Pavitra, Sathish Dayakshini, Verma Rama Shanker
Stem Cell and Molecular Biology Laboratory, Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras, Chennai, 600036 Tamilnadu India.
Department of Life Sciences, School of Basic Sciences and Research, Sharda University, Greater Noida, 201310 India.
3 Biotech. 2022 Jan;12(1):12. doi: 10.1007/s13205-021-03058-2. Epub 2021 Dec 9.
Mesenchymal stem cells (MSCs) differentiation toward cardiovascular lineage prediction using the global methylome profile will highlight its prospective utility in regenerative medicine. We examined the propensity prediction to cardiovascular lineage using 5-Aza, a well-known cardiac lineage inducer. The customized 180 K microarray was performed and further analysis of global differentially methylated regions by Ingenuity pathway analysis (IPA) in both MSCs and 5-AC-treated MSCs. The cluster enrichment tools sorted differentially enriched genes and further annotated to construct the interactive networks. Prediction analysis revealed pathways pertaining to the cardiovascular lineage found active in the native MSCs, suggesting its higher propensity to undergo cardiac, smooth muscle cell, and endothelial lineages in vitro. Interestingly, gene interaction network also proposed majorly stemness gene network and , cardiac-specific transcription factors , and were upregulated in the native MSCs. Furthermore, the expression of cardiovascular lineage specific markers such as , and various forms of (cardiac, sarcomeric, smooth muscle) were validated in native MSCs using real time PCR and immunostaining and blotting analysis. In 5-AC-treated MSCs, mosaic interactive networks were observed to persuade towards osteogenesis and cardiac lineage, indicating that 5-AC treatment resulted in nonspecific lineage induction in MSCs, while MSCs by default have a higher propensity to undergo cardiovascular lineage.
The online version contains supplementary material available at 10.1007/s13205-021-03058-2.
利用全基因组甲基化谱预测间充质干细胞（MSCs）向心血管谱系的分化将突出其在再生医学中的潜在效用。我们使用一种著名的心脏谱系诱导剂5-氮杂胞苷（5-Aza）来检测MSCs向心血管谱系的倾向预测。进行了定制的180K微阵列分析，并通过Ingenuity通路分析（IPA）对MSCs和5-氮杂胞苷处理的MSCs中的全基因组差异甲基化区域进行了进一步分析。聚类富集工具对差异富集基因进行分类，并进一步注释以构建交互网络。预测分析揭示了在天然MSCs中发现的与心血管谱系相关的通路，表明其在体外向心脏、平滑肌细胞和内皮谱系分化的倾向更高。有趣的是，基因相互作用网络还主要提出了干性基因网络，并且在天然MSCs中，心脏特异性转录因子被上调。此外，使用实时PCR、免疫染色和印迹分析在天然MSCs中验证了心血管谱系特异性标志物如、和各种形式的（心脏、肌节、平滑肌）的表达。在5-氮杂胞苷处理的MSCs中，观察到镶嵌式交互网络促使其向成骨和心脏谱系分化，表明5-氮杂胞苷处理导致MSCs发生非特异性谱系诱导，而MSCs默认情况下具有更高的向心血管谱系分化的倾向。
在线版本包含可在10.1007/s13205-021-03058-2获取的补充材料。
