Convergence of Notch and β-catenin signaling induces arterial fate in vascular progenitors

Molecular mechanisms controlling arterial–venous specification have not been fully elucidated. Previously, we established an embryonic stem cell differentiation system and demonstrated that activation of cAMP signaling together with VEGF induces arterial endothelial cells (ECs) from Flk1+ vascular progenitor cells. Here, we show novel arterial specification machinery regulated by Notch and β-catenin signaling. Notch and GSK3β-mediated β-catenin signaling were activated downstream of cAMP through phosphatidylinositol-3 kinase. Forced activation of Notch and β-catenin with VEGF completely reconstituted cAMP-elicited arterial EC induction, and synergistically enhanced target gene promoter activity in vitro and arterial gene expression during in vivo angiogenesis. A protein complex with RBP-J, the intracellular domain of Notch, and β-catenin was formed on RBP-J binding sites of arterial genes in arterial, but not venous ECs. This molecular machinery for arterial specification leads to an integrated and more comprehensive understanding of vascular signaling.

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Extracellular Matrix and Integrins in Embryonic Stem Cell Differentiation

Biochemistry Insights ◽

10.4137/bci.s30377 ◽

2015 ◽

Vol 8s2 ◽

pp. BCI.S30377 ◽

Cited By ~ 18

Author(s):

Han Wang ◽

Xie Luo ◽

Jake Leighton

Keyword(s):

Extracellular Matrix ◽

Pancreatic Beta Cells ◽

Embryonic Stem ◽

Stem Cell Differentiation ◽

Embryonic Stem Cell Differentiation ◽

In Vitro Differentiation ◽

Integrin Receptors

Embryonic stem cells (ESCs) are pluripotent cells with great therapeutic potentials. The in vitro differentiation of ESC was designed by recapitulating embryogenesis. Significant progress has been made to improve the in vitro differentiation protocols by toning soluble maintenance factors. However, more robust methods for lineage-specific differentiation and maturation are still under development. Considering the complexity of in vivo embryogenesis environment, extracellular matrix (ECM) cues should be considered besides growth factor cues. ECM proteins bind to cells and act as ligands of integrin receptors on cell surfaces. Here, we summarize the role of the ECM and integrins in the formation of three germ layer progenies. Various ECM–integrin interactions were found, facilitating differentiation toward definitive endoderm, hepatocyte-like cells, pancreatic beta cells, early mesodermal progenitors, cardiomyocytes, neuroectoderm lineages, and epidermal cells, such as keratinocytes and melanocytes. In the future, ECM combinations for the optimal ESC differentiation environment will require substantial study.

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Mouse lysocardiolipin acyltransferase controls the development of hematopoietic and endothelial lineages during in vitro embryonic stem-cell differentiation

Blood ◽

10.1182/blood-2007-04-086827 ◽

2007 ◽

Vol 110 (10) ◽

pp. 3601-3609 ◽

Cited By ~ 25

Author(s):

Chengyan Wang ◽

Patrick W. Faloon ◽

Zhijia Tan ◽

Yaxin Lv ◽

Pengbo Zhang ◽

...

Keyword(s):

Cell Differentiation ◽

Messenger Rna ◽

Molecular Mechanisms ◽

Embryonic Stem ◽

Stem Cell Differentiation ◽

Embryoid Bodies ◽

Embryonic Stem Cell Differentiation ◽

Hematopoietic Stem ◽

Endothelial Genes

Abstract The blast colony-forming cell (BL-CFC) was identified as an equivalent to the hemangioblast during in vitro embryonic stem (ES) cell differentiation. However, the molecular mechanisms underlying the generation of the BL-CFC remain largely unknown. Here we report the isolation of mouse lysocardiolipin acyltransferase (Lycat) based on homology to zebrafish lycat, a candidate gene for the cloche locus. Mouse Lycat is expressed in hematopoietic organs and is enriched in the Lin−C-Kit+Sca-1+ hematopoietic stem cells in bone marrow and in the Flk1+/hCD4+(Scl+) hemangioblast population in embryoid bodies. The forced Lycat transgene leads to increased messenger RNA expression of hematopoietic and endothelial genes as well as increased blast colonies and their progenies, endothelial and hematopoietic lineages. The Lycat small interfering RNA transgene leads to a decrease expression of hematopoietic and endothelial genes. An unbiased genomewide microarray analysis further substantiates that the forced Lycat transgene specifically up-regulates a set of genes related to hemangioblasts and hematopoietic and endothelial lineages. Therefore, mouse Lycat plays an important role in the early specification of hematopoietic and endothelial cells, probably acting at the level of the hemangioblast.

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Defining the earliest step of cardiovascular progenitor specification during embryonic stem cell differentiation

The Journal of Cell Biology ◽

10.1083/jcb.201007063 ◽

2011 ◽

Vol 192 (5) ◽

pp. 751-765 ◽

Cited By ~ 93

Author(s):

Antoine Bondue ◽

Simon Tännler ◽

Giuseppe Chiapparo ◽

Samira Chabab ◽

Mirana Ramialison ◽

...

Keyword(s):

Stem Cell ◽

Embryonic Stem Cell ◽

Fluorescent Protein ◽

Transcriptional Profiling ◽

Embryonic Stem ◽

Stem Cell Differentiation ◽

Embryonic Stem Cell Differentiation ◽

Heart Fields

During embryonic development and embryonic stem cell (ESC) differentiation, the different cell lineages of the mature heart arise from two types of multipotent cardiovascular progenitors (MCPs), the first and second heart fields. A key question is whether these two MCP populations arise from differentiation of a common progenitor. In this paper, we engineered Mesp1–green fluorescent protein (GFP) ESCs to isolate early MCPs during ESC differentiation. Mesp1-GFP cells are strongly enriched for MCPs, presenting the ability to differentiate into multiple cardiovascular lineages from both heart fields in vitro and in vivo. Transcriptional profiling of Mesp1-GFP cells uncovered cell surface markers expressed by MCPs allowing their prospective isolation. Mesp1 is required for MCP specification and the expression of key cardiovascular transcription factors. Isl1 is expressed in a subset of early Mesp1-expressing cells independently of Mesp1 and acts together with Mesp1 to promote cardiovascular differentiation. Our study identifies the early MCPs residing at the top of the cellular hierarchy of cardiovascular lineages during ESC differentiation.

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The Lysine Methylase SMYD3 Modulates Mesendodermal Commitment during Development

Cells ◽

10.3390/cells10051233 ◽

2021 ◽

Vol 10 (5) ◽

pp. 1233

Author(s):

Raffaella Fittipaldi ◽

Pamela Floris ◽

Valentina Proserpio ◽

Franco Cotelli ◽

Monica Beltrame ◽

...

Keyword(s):

Embryonic Stem ◽

Stem Cell Differentiation ◽

Model Systems ◽

Embryonic Stem Cell Differentiation ◽

In Vitro Differentiation ◽

Cancer Models ◽

Lineage Markers

SMYD3 (SET and MYND domain containing protein 3) is a methylase over-expressed in cancer cells and involved in oncogenesis. While several studies uncovered key functions for SMYD3 in cancer models, the SMYD3 role in physiological conditions has not been fully elucidated yet. Here, we dissect the role of SMYD3 at early stages of development, employing mouse embryonic stem cells (ESCs) and zebrafish as model systems. We report that SMYD3 depletion promotes the induction of the mesodermal pattern during in vitro differentiation of ESCs and is linked to an upregulation of cardiovascular lineage markers at later stages. In vivo, smyd3 knockdown in zebrafish favors the upregulation of mesendodermal markers during zebrafish gastrulation. Overall, our study reveals that SMYD3 modulates levels of mesendodermal markers, both in development and in embryonic stem cell differentiation.

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Sox7-sustained expression alters the balance between proliferation and differentiation of hematopoietic progenitors at the onset of blood specification

Blood ◽

10.1182/blood-2009-06-226290 ◽

2009 ◽

Vol 114 (23) ◽

pp. 4813-4822 ◽

Cited By ~ 42

Author(s):

Arnaud Gandillet ◽

Alicia G. Serrano ◽

Stella Pearson ◽

Michael Lie-A-Ling ◽

Georges Lacaud ◽

...

Keyword(s):

Molecular Mechanisms ◽

Embryonic Stem ◽

Global Gene Expression ◽

Stem Cell Differentiation ◽

Embryonic Stem Cell Differentiation ◽

Potential Candidate ◽

Hematopoietic Progenitors ◽

Potential Candidate Gene ◽

Proliferation And Differentiation

Abstract The molecular mechanisms that regulate the balance between proliferation and differentiation of precursors at the onset of hematopoiesis specification are poorly understood. By using a global gene expression profiling approach during the course of embryonic stem cell differentiation, we identified Sox7 as a potential candidate gene involved in the regulation of blood lineage formation from the mesoderm germ layer. In the present study, we show that Sox7 is transiently expressed in mesodermal precursors as they undergo specification to the hematopoietic program. Sox7 knockdown in vitro significantly decreases the formation of both primitive erythroid and definitive hematopoietic progenitors as well as endothelial progenitors. In contrast, Sox7-sustained expression in the earliest committed hematopoietic precursors promotes the maintenance of their multipotent and self-renewing status. Removal of this differentiation block driven by Sox7-enforced expression leads to the efficient differentiation of hematopoietic progenitors to all erythroid and myeloid lineages. This study identifies Sox7 as a novel and important player in the molecular regulation of the first committed blood precursors. Furthermore, our data demonstrate that the mere sustained expression of Sox7 is sufficient to completely alter the balance between proliferation and differentiation at the onset of hematopoiesis.

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The murine H19 gene is activated during embryonic stem cell differentiation in vitro and at the time of implantation in the developing embryo

Development ◽

10.1242/dev.113.4.1105 ◽

1991 ◽

Vol 113 (4) ◽

pp. 1105-1114 ◽

Cited By ~ 20

Author(s):

F. Poirier ◽

C.T. Chan ◽

P.M. Timmons ◽

E.J. Robertson ◽

M.J. Evans ◽

...

Keyword(s):

Embryonic Stem ◽

Stem Cell Differentiation ◽

Cell Types ◽

Embryoid Bodies ◽

Embryonic Stem Cell Differentiation ◽

Cdna Clones ◽

Stage Of Development ◽

Before And After

The differentiation in vitro of murine embryonic stem cells to embryoid bodies mimics events that occur in vivo shortly before and after embryonic implantation. We have used this system, together with differential cDNA cloning, to identify genes the expression of which is regulated during early embryogenesis. Here we describe the isolation of several such cDNA clones, one of which corresponds to the gene H19. This gene is activated in extraembryonic cell types at the time of implantation, suggesting that it may play a role at this stage of development, and is subsequently expressed in all of the cells of the mid-gestation embryo with the striking exception of most of those of the developing central and peripheral nervous systems. After birth, expression of this gene ceases or is dramatically reduced in all tissues.

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Diverse but unique astrocytic phenotypes during embryonic stem cell differentiation, culturing and aging

10.1101/2021.08.02.454573 ◽

2021 ◽

Author(s):

Kiara Freitag ◽

Pascale Eede ◽

Andranik Ivanov ◽

Shirin Schneeberger ◽

Tatiana Borodina ◽

...

Keyword(s):

Stem Cell ◽

Embryonic Stem Cell ◽

Embryonic Stem ◽

Calcium Signalling ◽

Stem Cell Differentiation ◽

Embryonic Stem Cell Differentiation ◽

The Central Nervous System ◽

Transcriptomic Level

Astrocytes are resident glia cells of the central nervous system (CNS) that play complex and heterogeneous roles in brain development, homeostasis and disease. Since their vast involvement in health and disease is becoming increasingly recognized, suitable and reliable tools for studying these cells in vivo and in vitro are of utmost importance. One of the key challenges hereby is to adequately mimic their context-dependent in vivo phenotypes and functions in vitro. To better understand the spectrum of astrocytic variations in defined settings we performed a side-by-side-comparison of embryonic stem cell (ESC)-derived astrocytes as well as primary neonatal and adult astrocytes, revealing major differences on a functional and transcriptomic level, specifically on proliferation, migration, calcium signalling and cilium activity. Our results highlight the need to carefully consider the choice of astrocyte origin and phenotype with respect to age, isolation and culture protocols based on the respective biological question.

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