当前位置: X-MOL 学术Stem Cells Dev. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
microRNA Sequencing of CD34+ Sorted Adipose Stem Cells Undergoing Endotheliogenesis
Stem Cells and Development ( IF 2.5 ) Pub Date : 2021-03-03 , DOI: 10.1089/scd.2020.0173
Shahensha Shaik 1 , Elizabeth Martin 2 , Daniel Hayes 3 , Jeffrey Gimble 4 , Ram Devireddy 1
Affiliation  

While several microRNAs (miRNAs) that regulate the endotheliogenesis and further promote angiogenesis have been identified in various cancers, the identification of miRNAs that can drive the differentiation of adipose derived stromal/stem cells (ASCs) into the endothelial lineage has been largely unexplored. In this study, CD34+ ASCs sorted using magnetic bead separation were induced to differentiate along the endothelial pathway. miRNA sequencing of ASCs at day 3, 9, and 14 of endothelial differentiation was performed on Ion Proton sequencing system. The data obtained by this high-throughput method were aligned to the human genome HG38, and the differentially expressed miRNAs during endothelial differentiation at various time points (day 3, 9, and 14) were identified. The gene targets of the identified miRNAs were obtained through miRWalk database. The network-pathway analysis of miRNAs and their targets was performed using the Database for Annotation, Visualization and Integrated Discovery (DAVID) bioinformatic tools to determine the potential candidate miRNAs that promote endothelial differentiation. Based on these analyses, six upregulated miRNAs (miR-181a-5p, miR-330-5p, miR-335-3p, miR-15b-5p, miR-99a-5p, and miR-199a-5p) and six downregulated miRNAs (miR-145-5p, miR-155-5p, miR-193a-3p, miR-125a-5p, miR-221-5p, and miR-222-3p) were chosen for further studies. In vitro evaluation of these miRNAs to induce endothelial differentiation when transfected into CD34+ sorted ASCs was studied using Von Willebrand Factor (VWF) staining and quantitative real time–polymerase chain reaction (qRT-PCR). Our results suggest that miRNAs: 335-5p, 330-5p, 181a-5p and anti-miRNAs: 125a-5p, 145-5p can likely induce endothelial differentiation in CD34+ sorted ASCs. Further studies are clearly required to elucidate the specific mechanisms on how miRNAs or anti-miRNAs identified through bioinformatics approach can induce the endotheliogenesis in ASCs.

中文翻译:


进行内皮生成的 CD34+ 分选脂肪干细胞的 microRNA 测序



虽然在各种癌症中已经鉴定出几种调节内皮生成并进一步促进血管生成的 microRNA (miRNA),但能够驱动脂肪源性基质/干细胞 (ASC) 分化为内皮谱系的 miRNA 的鉴定在很大程度上尚未被探索。在本研究中,利用磁珠分离分选的 CD34+ ASC 被诱导沿内皮途径分化。在 Ion Proton 测序系统上对内皮分化第 3、9 和 14 天的 ASC 进行 miRNA 测序。通过这种高通量方法获得的数据与人类基因组HG38进行比对,并鉴定了内皮分化过程中不同时间点(第3、9和14天)差异表达的miRNA。通过miRWalk数据库获得所鉴定的miRNA的基因靶标。使用注释、可视化和集成发现数据库 (DAVID) 生物信息学工具对 miRNA 及其靶标进行网络通路分析,以确定促进内皮分化的潜在候选 miRNA。基于这些分析,六种上调的 miRNA(miR-181a-5p、miR-330-5p、miR-335-3p、miR-15b-5p、miR-99a-5p 和 miR-199a-5p)和六种下调的 miRNA选择 (miR-145-5p、miR-155-5p、miR-193a-3p、miR-125a-5p、miR-221-5p 和 miR-222-3p) 进行进一步研究。使用冯维勒布兰德因子 (VWF) 染色和定量实时聚合酶链反应 (qRT-PCR) 对这些 miRNA 转染至 CD34+ 分选的 ASC 时诱导内皮分化的体外评估进行了研究。我们的结果表明,miRNA:335-5p、330-5p、181a-5p 和抗 miRNA:125a-5p、145-5p 可能会诱导 CD34+ 分选的 ASC 中的内皮分化。 显然需要进一步的研究来阐明通过生物信息学方法识别的 miRNA 或抗 miRNA 如何诱导 ASC 内皮细胞生成的具体机制。
更新日期:2021-03-05
down
wechat
bug