TLR4 expression in mouse embryonic stem cells and in stem cell-derived vascular cells is regulated by epigenetic modifications

AbstractMicroRNAs (miRNAs) are a class of non-coding small RNAs that function in almost every known cellular activity. MiRNAs play an important role in gene regulation that controls embryonic stem cell (ESC) pluripotency and differentiation, as well as induced pluripotent stem cell (iPSC) reprogramming. In this study, we identified nine novel miRNAs by mining the deep sequencing dataset from mouse embryonic stem cells, mouse embryonic fibroblasts (MEF) and three kinds of reprogrammed pluripotent cells. Most of them are non-conserved but species-specific and cell-specific miRNAs. Eight miRNAs are derived from gene introns, including a “mirtron” miRNA, miR-novel-41. We also showed that miR-novel-27 is a mouse-specific miRNA and the 5′ arm of its precursor hairpin, embedding the mature miR-novel-27, uniquely exists in mouse species but not in any other Placentalia animals. Notably, the 5′ arm of the pre-miR-novel-27 hairpin shows nearly perfect palindrome to the 3′ arm suggesting that it was generated by inverted duplication of the 3′ arm. By this mechanism, the pre-miR-novel-27 hairpin was de novo gained in the mouse genome. This is a new type of de novo miRNA emergence mechanism in animals, which we called “inverted local half hairpin duplication” here. In addition, very limited nucleotide mutants accumulated on the newly emerged 5′ arm since its birth suggesting an especially young evolutionary history of the miR-novel-27 gene.

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Assessment of Stem Cell Markers During Long-Term Culture of Mouse Embryonic Stem Cells

Cytotechnology ◽

10.1023/b:cyto.0000043414.90681.c2 ◽

2004 ◽

Vol 44 (1/2) ◽

pp. 77-91 ◽

Cited By ~ 24

Author(s):

A. Berrill ◽

H.L. Tan ◽

S.C. Wuang ◽

W.J. Fong ◽

Andre B.H. Choo ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Embryonic Stem Cells ◽

Embryonic Stem ◽

Mouse Embryonic Stem Cells ◽

Stem Cell Markers ◽

Cell Markers ◽

Long Term Culture

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Histone Acetyltransferase KAT2A Stabilises Pluripotency with Control of Transcriptional Heterogeneity

10.1101/347476 ◽

2018 ◽

Cited By ~ 1

Author(s):

Naomi Moris ◽

Shlomit Edri ◽

Denis Seyres ◽

Rashmi Kulkarni ◽

Ana Filipa Domingues ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Embryonic Stem Cells ◽

Cell Fate ◽

Regulatory Networks ◽

Environmental Changes ◽

Embryonic Stem ◽

Mouse Embryonic Stem Cells ◽

Histone Acetyl Transferase ◽

Unicellular Organisms

ABSTRACTCell fate transitions in mammalian stem cell systems have often been associated with transcriptional heterogeneity, however existing data have failed to establish a functional or mechanistic link between the two phenomena. Experiments in unicellular organisms support the notion that transcriptional heterogeneity can be used to facilitate adaptability to environmental changes and have identified conserved chromatin-associated factors that modulate levels of transcriptional noise. Herein, we show destabilisation of pluripotency-associated gene regulatory networks through increased transcriptional heterogeneity of mouse embryonic stem cells in which paradigmatic histone acetyl-transferase, and candidate noise modulator, Kat2a (yeast orthologue Gcn5) has been inhibited. Functionally, network destabilisation associates with reduced pluripotency and accelerated mesendodermal differentiation, with increased probability of transitions into lineage commitment. Thus, we functionally link transcriptional heterogeneity to cell fate transitions through manipulation of the histone acetylation landscape of mouse embryonic stem cells and establish a general paradigm that could be exploited in other normal and malignant stem cell fate transitions.

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BMP Signaling Is Crucial for Regulation Vascular Progenitor Development in Human Embryonic Stem Cells.

Blood ◽

10.1182/blood.v114.22.3037.3037 ◽

2009 ◽

Vol 114 (22) ◽

pp. 3037-3037

Author(s):

Zack Z. Wang ◽

Hao Bai ◽

YongXing Gao ◽

Melanie Arzigian ◽

Don M. Wojchowski ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Embryonic Stem Cells ◽

Progenitor Cells ◽

Human Embryonic Stem Cells ◽

Embryonic Stem ◽

Vascular Cells ◽

Cd34 Cells ◽

Vascular Progenitor Cells ◽

Tgfbeta Signaling

Abstract Abstract 3037 Poster Board II-1013 The generation of vascular cells from pluripotent stem cells, including human embryonic stem cells (hESCs) and induced pluripotent stem (iPS) cells, may facilitate tissue transplantation, reperfusion of ischemic tissues, and treatment of pathologies in which endothelial cell dysfunction exists. The signals that direct pluripotent stem cell differentiation into lineage-specific cells remain largely unknown. To identify vascular progenitor cells during hESC differentiation, we characterized various subpopulations that may differentiate into endothelial cells (ECs) and smooth muscle cells (SMCs). In our newly established serum-free medium, hESCs sequentially differentiated into CD34+CD31-, CD34+CD31+, and then CD34-CD31+ cells. Real-time PCR analysis indicated that EC- and SMC-specific genes, including VEGFR2, Tie2, VE-Cad, vWF, SMA, SM22-alpha, calponin, and caldesmon, were expressed significantly high in CD34+CD31+ cells. Furthermore, the CD34+CD31+ cells that were cultured in EC or SMC growth medium had high differentiation potential to become ECs and SMCs. These data suggested that CD34+CD31+ cells contain vascular progenitor cells. By treating hESCs with various factors at different time points, we found that BMP4 was critical to initiate hESC differentiation into CD34+CD31+ progenitor cells, whereas VEGF and FGF2 facilitated CD34+CD31+ cell development in the later stage. Conversely, TGFbeta promoted CD34+CD31- cells that were unable to give rise to ECs and SMCs. In addition, TGFbeta blocked the development of CD34+CD31+ cells induced by BMP4, suggesting that TGFbeta signaling negatively regulates the development of CD34+CD31+ cells during hESC differentiation. DNA microarray analysis indicated that a number of mesodermal genes were increased in BMP4-induced CD34+ cells, and a number of ectodermal genes were increased in TGFbeta-induced CD34+ cells. Therefore, the roles of BMP and TGFbeta signals in lineage differentiation may be inversely reflected in mesoderm and ectoderm. In addition, several pluripotent genes were expressed in TGFbeta-induced CD34+ cells, suggesting that TGFbeta signaling affects maintenance of stem cell pluripotency. The BMP-Smad inhibitor, dorsomorphin, inhibited phosphorylation of Smad1/5/8, and blocked hESC differentiation to CD34+CD31+ progenitor cells, suggesting that BMP Smad-dependent signaling is critical for CD34+CD31+ vascular progenitor development. Our findings provide new insight into how pluripotent hESCs differentiate to generate vascular cells. Disclosures No relevant conflicts of interest to declare.

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Slc25a36 modulates pluripotency of mouse embryonic stem cells by regulating mitochondrial function and glutathione level

Biochemical Journal ◽

10.1042/bcj20190057 ◽

2019 ◽

Vol 476 (11) ◽

pp. 1585-1604 ◽

Cited By ~ 1

Author(s):

Yanli Xin ◽

Yanliang Wang ◽

Liang Zhong ◽

Bingbo Shi ◽

Hui Liang ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Embryonic Stem Cells ◽

Mitochondrial Dysfunction ◽

Cell Fate ◽

Mitochondrial Function ◽

Embryonic Stem ◽

Mouse Embryonic Stem Cells ◽

Glutathione Metabolism ◽

Pluripotency Maintenance

Abstract Mitochondria play a central role in the maintenance of the naive state of embryonic stem cells. Many details of the mechanism remain to be fully elucidated. Solute carrier family 25 member 36 (Slc25a36) might regulate mitochondrial function through transporting pyrimidine nucleotides for mtDNA/RNA synthesis. Its physical role in this process remains unknown; however, Slc25a36 was recently found to be highly expressed in naive mouse embryonic stem cells (mESCs). Here, the function of Slc25a36 was characterized as a maintenance factor of mESCs pluripotency. Slc25a36 deficiency (via knockdown) has been demonstrated to result in mitochondrial dysfunction, which induces the differentiation of mESCs. The expression of key pluripotency markers (Pou5f1, Sox2, Nanog, and Utf1) decreased, while that of key TE genes (Cdx2, Gata3, and Hand1) increased. Cdx2-positive cells emerged in Slc25a36-deficient colonies under trophoblast stem cell culture conditions. As a result of Slc25a36 deficiency, mtDNA of knockdown cells declined, leading to impaired mitochondria with swollen morphology, decreased mitochondrial membrane potential, and low numbers. The key transcription regulators of mitochondrial biogenesis also decreased. These results indicate that mitochondrial dysfunction leads to an inability to support the pluripotency maintenance. Moreover, down-regulated glutathione metabolism and up-regulated focal adhesion reinforced and stabilized the process of differentiation by separately enhancing OCT4 degradation and promoting cell spread. This study improves the understanding of the function of Slc25a36, as well as the relationship of mitochondrial function with naive pluripotency maintenance and stem cell fate decision.

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