Downregulation of E-cadherin in pluripotent stem cells triggers partial EMT

Epithelial to mesenchymal transition (EMT) is a critical cellular process that has been well characterized during embryonic development and cancer metastasis and it also is implicated in several physiological and pathological events including embryonic stem cell differentiation. During early stages of differentiation, human embryonic stem cells pass through EMT where deeper morphological, molecular and biochemical changes occur. Though initially considered as a decision between two states, EMT process is now regarded as a fluid transition where cells exist on a spectrum of intermediate states. In this work, using a CRISPR interference system in human embryonic stem cells, we describe a molecular characterization of the effects of downregulation of E-cadherin, one of the main initiation events of EMT, as a unique start signal. Our results suggest that the decrease and delocalization of E-cadherin causes an incomplete EMT where cells retain their undifferentiated state while expressing several characteristics of a mesenchymal-like pheno-type. Namely, we found that E-cadherin downregulation induces SNAI1 and SNAI2 upregulation, promotes MALAT1 and LINC-ROR downregulation, modulates the expression of tight junction occludin 1 and gap junction connexin 43, increases human embryonic stem cells migratory capacity and delocalize b-catenin. Altogether, we believe our results provide a useful tool to model the molecular events of an unstable intermediate state and further identify multiple layers of molecular changes that occur during partial EMT.

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Downregulation of E-cadherin in pluripotent stem cells triggers partial EMT

Scientific Reports ◽

10.1038/s41598-021-81735-1 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

C. E. Aban ◽

A. Lombardi ◽

G. Neiman ◽

M. C. Biani ◽

A. La Greca ◽

...

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Human Embryonic Stem Cells ◽

Connexin 43 ◽

Cancer Metastasis ◽

Embryonic Stem ◽

Stem Cell Differentiation ◽

Embryonic Stem Cell Differentiation ◽

Mesenchymal Transition ◽

E Cadherin

AbstractEpithelial to mesenchymal transition (EMT) is a critical cellular process that has been well characterized during embryonic development and cancer metastasis and it also is implicated in several physiological and pathological events including embryonic stem cell differentiation. During early stages of differentiation, human embryonic stem cells pass through EMT where deeper morphological, molecular and biochemical changes occur. Though initially considered as a decision between two states, EMT process is now regarded as a fluid transition where cells exist on a spectrum of intermediate states. In this work, using a CRISPR interference system in human embryonic stem cells, we describe a molecular characterization of the effects of downregulation of E-cadherin, one of the main initiation events of EMT, as a unique start signal. Our results suggest that the decrease and delocalization of E-cadherin causes an incomplete EMT where cells retain their undifferentiated state while expressing several characteristics of a mesenchymal-like phenotype. Namely, we found that E-cadherin downregulation induces SNAI1 and SNAI2 upregulation, promotes MALAT1 and LINC-ROR downregulation, modulates the expression of tight junction occludin 1 and gap junction connexin 43, increases human embryonic stem cells migratory capacity and delocalize β-catenin. Altogether, we believe our results provide a useful tool to model the molecular events of an unstable intermediate state and further identify multiple layers of molecular changes that occur during partial EMT.

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Impact of AHR Ligand TCDD on Human Embryonic Stem Cells and Early Differentiation

International Journal of Molecular Sciences ◽

10.3390/ijms21239052 ◽

2020 ◽

Vol 21 (23) ◽

pp. 9052

Author(s):

Indrek Teino ◽

Antti Matvere ◽

Martin Pook ◽

Inge Varik ◽

Laura Pajusaar ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Embryonic Stem Cells ◽

Human Embryonic Stem Cells ◽

Target Genes ◽

Expression Patterns ◽

Embryonic Stem ◽

Stem Cell Differentiation ◽

Cell Types ◽

Early Differentiation

Aryl hydrocarbon receptor (AHR) is a ligand-activated transcription factor, which mediates the effects of a variety of environmental stimuli in multiple tissues. Recent advances in AHR biology have underlined its importance in cells with high developmental potency, including pluripotent stem cells. Nonetheless, there is little data on AHR expression and its role during the initial stages of stem cell differentiation. The purpose of this study was to investigate the temporal pattern of AHR expression during directed differentiation of human embryonic stem cells (hESC) into neural progenitor, early mesoderm and definitive endoderm cells. Additionally, we investigated the effect of the AHR agonist 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on the gene expression profile in hESCs and differentiated cells by RNA-seq, accompanied by identification of AHR binding sites by ChIP-seq and epigenetic landscape analysis by ATAC-seq. We showed that AHR is differentially regulated in distinct lineages. We provided evidence that TCDD alters gene expression patterns in hESCs and during early differentiation. Additionally, we identified novel potential AHR target genes, which expand our understanding on the role of this protein in different cell types.

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Connexin 43 is involved in early differentiation of human embryonic stem cells

Differentiation ◽

10.1016/j.diff.2018.12.003 ◽

2019 ◽

Vol 105 ◽

pp. 33-44 ◽

Cited By ~ 6

Author(s):

Qian Peng ◽

Chaomin Yue ◽

Andy Chun Hang Chen ◽

Kai Chuen Lee ◽

Sze Wan Fong ◽

...

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Human Embryonic Stem Cells ◽

Connexin 43 ◽

Embryonic Stem ◽

Early Differentiation

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Regulated expression of the small nuclear ribonucleoprotein particle protein SmN in embryonic stem cell differentiation.

Molecular and Cellular Biology ◽

10.1128/mcb.10.12.6817 ◽

1990 ◽

Vol 10 (12) ◽

pp. 6817-6820 ◽

Cited By ~ 15

Author(s):

N G Sharpe ◽

D G Williams ◽

D S Latchman

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Cell Lines ◽

Embryonal Carcinoma ◽

Embryonic Stem ◽

Stem Cell Differentiation ◽

Embryonic Stem Cell Differentiation ◽

Embryonal Carcinoma Cell ◽

Small Nuclear Ribonucleoprotein ◽

Nuclear Ribonucleoprotein

The SmN protein is a component of small nuclear ribonucleoprotein particles and is closely related to the ubiquitous SmB and B' splicing proteins. It is expressed in a limited range of tissues and cell types, including several undifferentiated embryonal carcinoma cell lines and undifferentiated embryonic stem cells. The protein declines to undetectable levels when embryonal carcinoma or embryonic stem cells are induced to differentiate, producing primitive endoderm or parietal endoderm or yielding embryonal bodies. This decline is due to a corresponding decrease in the level of the SmN mRNA. The potential role of SmN in the regulation of alternative splicing in embryonic cell lines and early embryos is discussed.

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A Unique Interplay between Rap1 and E-Cadherin in the Endocytic Pathway Regulates Self-Renewal of Human Embryonic Stem Cells

Stem Cells ◽

10.1002/stem.289 ◽

2009 ◽

pp. N/A-N/A ◽

Cited By ~ 6

Author(s):

Li Li ◽

Shuai Wang ◽

Anna Jezierski ◽

Lilian Moalim-Nour ◽

Kanishka Mohib ◽

...

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Human Embryonic Stem Cells ◽

Embryonic Stem ◽

Endocytic Pathway ◽

Self Renewal ◽

E Cadherin

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Epithelial–mesenchymal transition process in human embryonic stem cells cultured in feeder-free conditions

MHR Basic science of reproductive medicine ◽

10.1093/molehr/gal091 ◽

2006 ◽

Vol 13 (1) ◽

pp. 21-32 ◽

Cited By ~ 87

Author(s):

U. Ullmann ◽

P. In’t Veld ◽

C. Gilles ◽

K. Sermon ◽

M. De Rycke ◽

...

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Human Embryonic Stem Cells ◽

Epithelial Mesenchymal Transition ◽

Embryonic Stem ◽

Transition Process ◽

Mesenchymal Transition ◽

Cells Cultured

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Human Embryonic Stem Cells Maintain Pluripotency after E‐Cadherin Expression Knockdown

The FASEB Journal ◽

10.1096/fasebj.24.1_supplement.lb172 ◽

2010 ◽

Vol 24 (S1) ◽

Author(s):

Shanel Nicole Byas ◽

Joseph Klim ◽

Laura Kiessling

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Human Embryonic Stem Cells ◽

Embryonic Stem ◽

E Cadherin

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323 ESTROGEN-INDUCED EPITHELIAL-MESENCHYMAL TRANSITION AND EXPRESSION OF PROAPOPTOTIC MARKERS IN PORCINE STEM CELLS

Reproduction Fertility and Development ◽

10.1071/rdv27n1ab323 ◽

2015 ◽

Vol 27 (1) ◽

pp. 250

Author(s):

Y.-S. Kim ◽

S.-H. Hyun ◽

C.-K. Lee ◽

K.-C. Choi

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Epithelial Mesenchymal Transition ◽

Steroid Hormone ◽

Embryonic Stem ◽

Transgenic Pig ◽

Rt Pcr ◽

Mesenchymal Transition ◽

Apoptotic Markers ◽

E Cadherin

In transgenic pig production for generating animal models of human diseases, apoptosis of early implantation embryo disturbs the transgenic pig production. In general, epithelial-mesenchymal transition (EMT) is considered important in embryo development and apoptosis. In addition, it was reported that 17β-oestradiol (E2), among hormones that participate in early implantation of embryo, could induce EMT and neural differentiation in mouse embryonic stem cells. Therefore, in this study, we examined the effects of the steroid hormone, E2, in the changes of EMT and apoptotic markers in porcine embryonic stem cells (pESC) and porcine induced pluripotent stem cells (piPSC). During the study, we cultured pESC and piPSC in pESC media containing basic fibroblast growth factor (b-FGF) and leukemia inhibitory factor (LIF) and performed RT-PCR and an alkaline phophatase (AP) test to measure pluripotent and undifferentiation markers of these porcine stem cells. The RT-PCR results showed that OCT4, NANOG, and SOX2 were expressed in these pESC and piPSC, indicating their pluripotency as stem cells. Also, these porcine stem cells showed positive AP activity, demonstrating undifferentiation. Additionally, we treated pESC and piPSC with E2 to examine effects of steroid hormone on the changes of EMT and apoptotic markers (i.e. bcl-2, bax, E-cad, and vimentin). The E2 treatment increased the expression of vimentin and bcl-2, while decreased the expression of E-cadherin and bax. By using immunocytochemistry (ICC), we examined the protein expression of EMT markers, which are vimentin and E-cadherin at the translational level, and found that expression of vimentin protein was increased while E-cadherin protein level was reduced at periphery of the colonies in pESC and piPSC. In conclusion, these results indicate that E2 can promote EMT process and reverse apoptosis in these pESC and iPSC. In a future study, we will further examine the effects of progesterone on the expressions of EMT and apoptotic markers in pESC and piPSC. Consequently, this study will contribute to elucidate the underlying mechanisms of EMT and apoptosis controlled by steroid hormones in porcine stem cells.

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