Intrinsic transition of embryonic stem-cell differentiation into neural progenitors

Kamiya, Daisuke; Banno, Satoe; Sasai, Noriaki; Ohgushi, Masatoshi; Inomata, Hajime; Watanabe, Kiichi; Kawada, Manabu; Yakura, Rieko; Kanazawa, Hiroshi; Nakao, Kazuki; Jakt, Lars Martin; Nishikawa, Shin‐Ichi; Sasai, Yoshiki

doi:10.1038/nature09726

Cited by 192 publications

(194 citation statements)

References 35 publications

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“…4A). In mouse ESC neural differentiation, the neural fate commitment begins when NPC-specific gene Sox1 starts to express (30). By comparing with the microarray data of in vitro mouse ESC neural differentiation, we noticed that day 8 of Module 3 was highly correlated with the day 3 Sox1 ϩ cells, indicating the initiation of mouse neural fate (Fig.…”

Section: Comparative Analysis Validates Module 3 Associated Day 8/10 mentioning

confidence: 99%

Transcriptome analysis reveals determinant stages controlling human embryonic stem cell commitment to neuronal cells

Wang

Qiao³

et al. 2017

Journal of Biological Chemistry

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Edited by Xiao-Fan WangProper neural commitment is essential for ensuring the appropriate development of the human brain and for preventing neurodevelopmental diseases such as autism spectrum disorders, schizophrenia, and intellectual disorders. However, the molecular mechanisms underlying the neural commitment in humans remain elusive. Here, we report the establishment of a neural differentiation system based on human embryonic stem cells (hESCs) and on comprehensive RNA sequencing analysis of transcriptome dynamics during early hESC differentiation. Using weighted gene co-expression network analysis, we reveal that the hESC neurodevelopmental trajectory has five stages: pluripotency (day 0); differentiation initiation (days 2, 4, and 6); neural commitment (days 8 -10); neural progenitor cell proliferation (days 12, 14, and 16); and neuronal differentiation (days 18, 20, and 22). These stages were characterized by unique module genes, which may recapitulate the early human cortical development. Moreover, a comparison of our RNA-sequencing data with several other transcriptome profiling datasets from mice and humans indicated that Module 3 associated with the day 8 -10 stage is a critical window of fate switch from the pluripotency to the neural lineage. Interestingly, at this stage, no key extrinsic signals were activated. In contrast, using CRISPR/ Cas9 -mediated gene knockouts, we also found that intrinsic hub transcription factors, including the schizophrenia-associated SIX3 gene and septo-optic dysplasia-related HESX1 gene, are required to program hESC neural determination. Our results improve the understanding of the mechanism of neural commitment in the human brain and may help elucidate the etiology of human mental disorders and advance therapies for managing these conditions. Brain development is one of the most complicated and hierarchical events in mammals. A series of temporal processes, including neural commitment, patterning and sub-regionalization, neurogenesis, and neuronal network formation, are required to generate a functional brain (1-4). Defects in any of these processes will lead to neurodevelopmental diseases such as autism spectrum disorders, schizophrenia, depression, and epilepsy. Neural commitment occurs after gastrulation and initiates the program to form neural epithelium (5-7). Many studies on amphibians and rodents reveal that the inhibition of BMP 3 signaling is an evolutionally conserved mechanism for neural induction; in addition, intrinsic factors play pivotal roles to direct fate transition from pluripotency to neural lineage (8 -11). Nonetheless, how the neural commitment is ensured in human brain has not been clearly elucidated. Systematically understanding the critical role of signaling pathways and transcriptional factors in modulating and shaping human brain development is hindered by limited accessibility to early embryos and inadequate amounts of stage-specific and cell type-specific materials. These problems may now be solved by the use of human embryonic stem cells (h...

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Section: Comparative Analysis Validates Module 3 Associated Day 8/10 mentioning

confidence: 99%

Transcriptome analysis reveals determinant stages controlling human embryonic stem cell commitment to neuronal cells

Wang

Qiao³

et al. 2017

Journal of Biological Chemistry

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“…Population fate maps at early somite stages have identified two regions containing NMPs: the dorsal layer of the node-streak border (NSB) and the caudal lateral epiblast (CLE) on either side of the primitive streak (PS) (24)(25)(26)(27). In vitro, the induction of the neuroectoderm in ES cell (ESC) cultures is often referred to as the "default" pathway, because neuroectoderm readily develops in cultures that contain no serum or lack signaling through the Wnt, bone morphogenetic protein 4 (Bmp4), or activin signaling pathways (28)(29)(30).…”

Section: Significancementioning

confidence: 99%

Section: Tet3 Regulates the Balance Between Neuroectoderm And Cardiacmentioning

confidence: 99%

“…To investigate the role of Tet3 during neuroectoderm differentiation, we used an efficient system for in vitro neural differentiation of mESCs: serum-free floating cultures of embryoid-body like aggregates, referred to as "SFEB" (29,31). In SFEB cultures, mESCs undergo neuroectoderm differentiation by default, without the induction of mesodermal and endodermal differentiation (29,31). Consistent with our previous observation during retinoic acid-induced mESC differentiation (32), the expression levels of Tet1-3 displayed distinct patterns upon neural differentiation: Tet1 mRNA declined rapidly; Tet2 mRNA was initially maintained at a relatively steady level, then decreased on day 4, but recovered on day 6; and Tet3 mRNA showed a progressive increase, with >40-fold up-regulation on day 6 of SFEB culture compared with the starting mESCs (Fig.…”

Section: Tet3 Regulates the Balance Between Neuroectoderm And Cardiacmentioning

confidence: 99%

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Tet proteins influence the balance between neuroectodermal and mesodermal fate choice by inhibiting Wnt signaling

Yue

Pastor

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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TET-family dioxygenases catalyze conversion of 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC) and oxidized methylcytosines in DNA. Here, we show that mouse embryonic stem cells (mESCs), either lacking Tet3 alone or with triple deficiency of Tet1/2/3, displayed impaired adoption of neural cell fate and concomitantly skewed toward cardiac mesodermal fate. Conversely, ectopic expression of Tet3 enhanced neural differentiation and limited cardiac mesoderm specification. Genome-wide analyses showed that Tet3 mediates cell-fate decisions by inhibiting Wnt signaling, partly through promoter demethylation and transcriptional activation of the Wnt inhibitor secreted frizzled-related protein 4 (Sfrp4). Tet1/2/3-deficient embryos (embryonic day 8.0–8.5) showed hyperactivated Wnt signaling, as well as aberrant differentiation of bipotent neuromesodermal progenitors (NMPs) into mesoderm at the expense of neuroectoderm. Our data demonstrate a key role for TET proteins in modulating Wnt signaling and establishing the proper balance between neural and mesodermal cell fate determination in mouse embryos and ESCs.

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“…Although NKX2.1 7 and PAX8 8 are individually expressed in a variety of tissues and cell types, their co-expression is restricted to cells committed to differentiate into TFC. Induced overexpression of defined transcription factors has been shown to have a directing effect on the differentiation of Embryonic Stem Cells (ESCs) into specific cell types [9][10][11] . Despite the success of this experimental approach for cell differentiation or reprogramming, protocols promoting coordinated self-assembly of differentiated cells into distinct morphological units with functional properties reminiscent of organs and tissues in vivo [12][13][14] are still very sparse.…”

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confidence: 99%

Generation of Functional Thyroid from Embryonic Stem Cells

Antonica¹

2015

Thyroid Diseases in Childhood

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The primary function of thyroid gland is to metabolize iodide by synthesizing thyroid hormones that are critical regulators of growth, development and metabolism in virtually all tissues. To date, research on thyroid morphogenesis was missing an efficient stem-cell model system which allows to recapitulate in vitro the molecular and morphogenic events regulating thyroid follicular cells differentiation and subsequent assembly into functional thyroid follicles. Here we report that a transient overexpression of the transcription factors NKX2.1 and PAX8 is sufficient to direct mouse embryonic stem-cells (mESC) differentiation into thyroid follicular cells which organized into three-dimensional follicular structures when treated with thyrotropin. Those in vitro derived follicles showed significant iodide organification activity. Importantly, when grafted in vivo into athyreoid mice, these follicles rescued thyroid hormone plasma levels and promoted subsequent symptomatic recovery. Thus, mESC can be induced to differentiate into thyroid follicular cells in vitro and generate functional thyroid tissue.

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Intrinsic transition of embryonic stem-cell differentiation into neural progenitors

Cited by 192 publications

References 35 publications

Transcriptome analysis reveals determinant stages controlling human embryonic stem cell commitment to neuronal cells

Transcriptome analysis reveals determinant stages controlling human embryonic stem cell commitment to neuronal cells

Tet proteins influence the balance between neuroectodermal and mesodermal fate choice by inhibiting Wnt signaling

Generation of Functional Thyroid from Embryonic Stem Cells

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