RNA-Seq Based Transcriptome Analysis of Endothelial Differentiation of Bone Marrow Mesenchymal Stem Cells

Chengen Wang; Huihui Liu; Min Yang; Yun Bai; Hanyun Ren; Yinghua Zou; Ziping Yao; Bihui Zhang; Yuan Li

doi:10.1016/j.ejvs.2019.11.003

RNA-Seq Based Transcriptome Analysis of Endothelial Differentiation of Bone Marrow Mesenchymal Stem Cells

Eur J Vasc Endovasc Surg. 2020 May;59(5):834-842. doi: 10.1016/j.ejvs.2019.11.003. Epub 2019 Dec 23.

Authors

Chengen Wang¹, Huihui Liu², Min Yang³, Yun Bai⁴, Hanyun Ren², Yinghua Zou³, Ziping Yao³, Bihui Zhang³, Yuan Li⁵

Affiliations

¹ Department of Interventional Radiology and Vascular Surgery, Peking University First Hospital, Beijing, China; Department of Minimally Invasive Tumour Therapies Centre, Beijing Hospital, National Centre of Gerontology, Beijing, China.
² Department of Haematology, Peking University First Hospital, Beijing, China.
³ Department of Interventional Radiology and Vascular Surgery, Peking University First Hospital, Beijing, China.
⁴ Department of Cell Biology, School of Basic Medical Sciences, Peking University Health Science Centre, Beijing, China.
⁵ Department of Haematology, Peking University First Hospital, Beijing, China. Electronic address: drliyuan75@163.com.

PMID: 31874808
DOI: 10.1016/j.ejvs.2019.11.003

Abstract

Objective: The aim was to identify the change in gene expression between mesenchymal stem cells (MSCs) and induced endothelial cells (ECs) and to investigate the potential mechanism of endothelial differentiation based on ribonucleic acid sequencing (RNA-Seq) analysis.

Methods: MSCs were isolated from bone marrow and exposed to inducing medium. The dynamic transcription profiles of MSCs were identified and ECs were induced through RNA-seq. Differentially expressed genes (DEGs) were identified. Enrichment of functions and signalling pathways analysis were performed based on Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. Quantitative real time polymerase chain reaction (qRT-PCR) was used to validate the genes selected from RNA-Seq.

Results: In total, 2769 DEGs were identified, of which 1117 genes were upregulated and 1652 genes were downregulated. GO and KEGG pathway analyses identified significantly enriched pathways in DEGs, including extracellular matrix organisation, blood vessel morphogenesis, angiogenesis, extracellular matrix binding, growth factor binding and glycosaminoglycan binding extracellular matrix-receptor interaction pathway, cytokine-receptor interaction pathway and transforming growth factor (TGF)-β signalling pathway. All genes found to be associated with the TGF-β pathway were significantly downregulated. Eleven novel genes were also identified that most likely are involved in endothelial differentiation and were upregulated with more than 10 fold change, which were further validated by qRT-PCR.

Conclusion: The GO and KEGG analysis revealed that extracellular matrix, cytokines and the TGF-β pathway play an important role in the process of endothelial differentiation. Furthermore, 11 genes were found that may be involved in the differentiation of MSCs into ECs and contribute to current understanding of the differentiation mechanism.

Keywords: Endothelial cells; Endothelial differentiation; Gene expression; Stem cells.

MeSH terms

Cell Differentiation / genetics*
Endothelial Cells / cytology*
Humans
Mesenchymal Stem Cells / cytology*
RNA-Seq*