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RNA-seq reveals tight junction-relevant erythropoietic fate induced by OCT4 in human hair follicle mesenchymal stem cells.
Stem Cell Res Ther. 2020 Oct 27; 11(1):454.SC

Abstract

BACKGROUND

Human hair follicle mesenchymal stem cells (hHFMSCs) isolated from hair follicles possess multilineage differentiation potential. OCT4 is a gene critically associated with pluripotency properties. The cell morphology and adhesion of hHFMSCs significantly changed after transduction of OCT4 and two subpopulations emerged, including adherent cells and floating cell. Floating cells cultured in hematopoietic induction medium and stimulated with erythropoetic growth factors could transdifferentiate into mature erythrocytes, whereas adherent cells formed negligible hematopoietic colonies. The aim of this study was to reveal the role of cell morphology and adhesion on erythropoiesis induced by OCT4 in hHFMSCs and to characterize the molecular mechanisms involved.

METHODS

Floating cell was separated from adherent cell by centrifugation of the upper medium during cell culture. Cell size was observed through flow cytometry and cell adhesion was tested by disassociation and adhesion assays. RNA sequencing was performed to detect genome-wide transcriptomes and identify differentially expressed genes. GO enrichment analysis and KEGG pathway analysis were performed to analysis the functions and pathways enriched by differentially expressed genes. The expression of tight junction core members was verified by qPCR and Western blot. A regulatory network was constructed to figure out the relationship between cell adhesin, cytoskeleton, pluripotency, and hematopoiesis.

RESULTS

The overexpression of OCT4 influenced the morphology and adhesion of hHFMSCs. Transcripts in floating cells and adherent cells are quite different. Data analysis showed that upregulated genes in floating cells were mainly related to pluripotency, germ layer development (including hematopoiesis lineage development), and downregulated genes were mainly related to cell adhesion, cell junctions, and the cytoskeleton. Most molecules of the tight junction (TJ) pathway were downregulated and molecular homeostasis of the TJ was disturbed, as CLDNs were disrupted, and JAMs and TJPs were upregulated. The dynamic expression of cell adhesion-related gene E-cadherin and cytoskeleton-related gene ACTN2 might cause different morphology and adhesion. Finally, a regulatory network centered to OCT4 was constructed, which elucidated that he TJ pathway critically bridges pluripotency and hematopoiesis in a TJP1-dependent way.

CONCLUSIONS

Regulations of cell morphology and adhesion via the TJ pathway conducted by OCT4 might modulate hematopoiesis in hHFMSCs, thus developing potential mechanism of erythropoiesis in vitro.

Authors+Show Affiliations

Department of Pathology, College of Basic Medical Sciences, Qingdao University, 308 Ningxia Road, Qingdao, 266071, Shandong, China.Department of Critical Care Medicine, Qingdao Center Hospital, affiliated with Qingdao University, 127 Siliunan Road, Qingdao, 266042, Shandong, China.Department of Pathology, Qingdao Municipal Hospital, affiliated with Qingdao University, 1 Jiaozhou Road, Qingdao, 266011, Shandong, China.Department of Pathology, College of Basic Medical Sciences, Qingdao University, 308 Ningxia Road, Qingdao, 266071, Shandong, China. Department of Pathology, Qingdao Municipal Hospital, affiliated with Qingdao University, 1 Jiaozhou Road, Qingdao, 266011, Shandong, China.Department of Pathology, Qingdao Municipal Hospital, affiliated with Qingdao University, 1 Jiaozhou Road, Qingdao, 266011, Shandong, China.Department of Pathology, College of Basic Medical Sciences, Qingdao University, 308 Ningxia Road, Qingdao, 266071, Shandong, China.Department of Pathology, Qingdao Municipal Hospital, affiliated with Qingdao University, 1 Jiaozhou Road, Qingdao, 266011, Shandong, China.Department of Pathology, College of Basic Medical Sciences, Qingdao University, 308 Ningxia Road, Qingdao, 266071, Shandong, China.Department of Pathology, Qingdao Municipal Hospital, affiliated with Qingdao University, 1 Jiaozhou Road, Qingdao, 266011, Shandong, China. xiaojing_906@163.com.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

33109258

Citation

Yu, Xiaozhen, et al. "RNA-seq Reveals Tight Junction-relevant Erythropoietic Fate Induced By OCT4 in Human Hair Follicle Mesenchymal Stem Cells." Stem Cell Research & Therapy, vol. 11, no. 1, 2020, p. 454.
Yu X, Sun P, Huang X, et al. RNA-seq reveals tight junction-relevant erythropoietic fate induced by OCT4 in human hair follicle mesenchymal stem cells. Stem Cell Res Ther. 2020;11(1):454.
Yu, X., Sun, P., Huang, X., Chen, H., Huang, W., Ruan, Y., Jiang, W., Tan, X., & Liu, Z. (2020). RNA-seq reveals tight junction-relevant erythropoietic fate induced by OCT4 in human hair follicle mesenchymal stem cells. Stem Cell Research & Therapy, 11(1), 454. https://doi.org/10.1186/s13287-020-01976-1
Yu X, et al. RNA-seq Reveals Tight Junction-relevant Erythropoietic Fate Induced By OCT4 in Human Hair Follicle Mesenchymal Stem Cells. Stem Cell Res Ther. 2020 Oct 27;11(1):454. PubMed PMID: 33109258.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - RNA-seq reveals tight junction-relevant erythropoietic fate induced by OCT4 in human hair follicle mesenchymal stem cells. AU - Yu,Xiaozhen, AU - Sun,Pengpeng, AU - Huang,Xingang, AU - Chen,Hua, AU - Huang,Weiqing, AU - Ruan,Yingchun, AU - Jiang,Weina, AU - Tan,Xiaohua, AU - Liu,Zhijing, Y1 - 2020/10/27/ PY - 2020/07/23/received PY - 2020/10/14/accepted PY - 2020/10/28/entrez PY - 2020/10/29/pubmed PY - 2020/10/29/medline KW - Hematopoiesis KW - Human hair follicle mesenchymal stem cells KW - OCT4 KW - Tight junction pathway SP - 454 EP - 454 JF - Stem cell research & therapy JO - Stem Cell Res Ther VL - 11 IS - 1 N2 - BACKGROUND: Human hair follicle mesenchymal stem cells (hHFMSCs) isolated from hair follicles possess multilineage differentiation potential. OCT4 is a gene critically associated with pluripotency properties. The cell morphology and adhesion of hHFMSCs significantly changed after transduction of OCT4 and two subpopulations emerged, including adherent cells and floating cell. Floating cells cultured in hematopoietic induction medium and stimulated with erythropoetic growth factors could transdifferentiate into mature erythrocytes, whereas adherent cells formed negligible hematopoietic colonies. The aim of this study was to reveal the role of cell morphology and adhesion on erythropoiesis induced by OCT4 in hHFMSCs and to characterize the molecular mechanisms involved. METHODS: Floating cell was separated from adherent cell by centrifugation of the upper medium during cell culture. Cell size was observed through flow cytometry and cell adhesion was tested by disassociation and adhesion assays. RNA sequencing was performed to detect genome-wide transcriptomes and identify differentially expressed genes. GO enrichment analysis and KEGG pathway analysis were performed to analysis the functions and pathways enriched by differentially expressed genes. The expression of tight junction core members was verified by qPCR and Western blot. A regulatory network was constructed to figure out the relationship between cell adhesin, cytoskeleton, pluripotency, and hematopoiesis. RESULTS: The overexpression of OCT4 influenced the morphology and adhesion of hHFMSCs. Transcripts in floating cells and adherent cells are quite different. Data analysis showed that upregulated genes in floating cells were mainly related to pluripotency, germ layer development (including hematopoiesis lineage development), and downregulated genes were mainly related to cell adhesion, cell junctions, and the cytoskeleton. Most molecules of the tight junction (TJ) pathway were downregulated and molecular homeostasis of the TJ was disturbed, as CLDNs were disrupted, and JAMs and TJPs were upregulated. The dynamic expression of cell adhesion-related gene E-cadherin and cytoskeleton-related gene ACTN2 might cause different morphology and adhesion. Finally, a regulatory network centered to OCT4 was constructed, which elucidated that he TJ pathway critically bridges pluripotency and hematopoiesis in a TJP1-dependent way. CONCLUSIONS: Regulations of cell morphology and adhesion via the TJ pathway conducted by OCT4 might modulate hematopoiesis in hHFMSCs, thus developing potential mechanism of erythropoiesis in vitro. SN - 1757-6512 UR - https://www.unboundmedicine.com/medline/citation/33109258/RNA-seq_reveals_tight_junction-relevant_erythropoietic_fate_induced_by_OCT4_in_human_hair_follicle_mesenchymal_stem_cells L2 - https://stemcellres.biomedcentral.com/articles/10.1186/s13287-020-01976-1 DB - PRIME DP - Unbound Medicine ER -
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