Multiple roles of Activin/Nodal, bone morphogenetic protein, fibroblast growth factor and Wnt/β-catenin signalling in the anterior neural patterning of adherent human embryonic stem cell cultures

Lupo, Giuseppe; Novorol, Claire; Smith, Joseph R.; Vallier, Ludovic; Miranda, Elena; Alexander, Morgan R.; Biagioni, Stefano; Pedersen, Roger A.; Harris, William A.

doi:10.1098/rsob.120167

Cited by 34 publications

(55 citation statements)

References 57 publications

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“…For instance, induction of human neural tube cells posterior to the forebrain has been shown to require endogenous WNT ligand activity (Lupo et al, 2013). Also, retinoic acid/FGF activity can posteriorize human neural progenitors, consistent with the roles proposed for these signaling pathways in animal models.…”

Section: Discussionmentioning

confidence: 59%

WNT/β-catenin signaling mediates human neural crest induction via a pre-neural border intermediate

et al. 2016

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Neural crest (NC) cells arise early in vertebrate development, migrate extensively and contribute to a diverse array of ectodermal and mesenchymal derivatives. Previous models of NC formation suggested derivation from neuralized ectoderm, via meso-ectodermal, or neural-non-neural ectoderm interactions. Recent studies using bird and amphibian embryos suggest an earlier origin of NC, independent of neural and mesodermal tissues. Here, we set out to generate a model in which to decipher signaling and tissue interactions involved in human NC induction. Our novel human embryonic stem cell (ESC)-based model yields high proportions of multipotent NC cells (expressing SOX10, PAX7 and TFAP2A) in 5 days. We demonstrate a crucial role for WNT/β-catenin signaling in launching NC development, while blocking placodal and surface ectoderm fates. We provide evidence of the delicate temporal effects of BMP and FGF signaling, and find that NC development is separable from neural and/ or mesodermal contributions. We further substantiate the notion of a neural-independent origin of NC through PAX6 expression and knockdown studies. Finally, we identify a novel pre-neural border state characterized by early WNT/β-catenin signaling targets that displays distinct responses to BMP and FGF signaling from the traditional neural border genes. In summary, our work provides a fast and efficient protocol for human NC differentiation under signaling constraints similar to those identified in vivo in model organisms, and strengthens a framework for neural crest ontogeny that is separable from neural and mesodermal fates.

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Section: Discussionmentioning

confidence: 59%

WNT/β-catenin signaling mediates human neural crest induction via a pre-neural border intermediate

et al. 2016

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“…Recent reports have described activation of endogenous Wnt/β-catenin signaling during neural conversion of hESCs by TGFβ inhibition [52, 88]. As shown by treatments with Wnt/β-catenin inhibitors, this endogenous Wnt/β-catenin activation restrained induction of rostral forebrain fates in favor of caudal forebrain and/or midbrain fates [52, 88].…”

Section: Forebrain Specification In Neuralized Escs: Protecting Neuramentioning

confidence: 99%

From pluripotency to forebrain patterning: an in vitro journey astride embryonic stem cells

Lupo

Bertacchi

Carucci

et al. 2014

Cell. Mol. Life Sci.

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Embryonic stem cells (ESCs) have been used extensively as in vitro models of neural development and disease, with special efforts towards their conversion into forebrain progenitors and neurons. The forebrain is the most complex brain region, giving rise to several fundamental structures, such as the cerebral cortex, the hypothalamus, and the retina. Due to the multiplicity of signaling pathways playing different roles at distinct times of embryonic development, the specification and patterning of forebrain has been difficult to study in vivo. Research performed on ESCs in vitro has provided a large body of evidence to complement work in model organisms, but these studies have often been focused more on cell type production than on cell fate regulation. In this review, we systematically reassess the current literature in the field of forebrain development in mouse and human ESCs with a focus on the molecular mechanisms of early cell fate decisions, taking into consideration the specific culture conditions, exogenous and endogenous molecular cues as described in the original studies. The resulting model of early forebrain induction and patterning provides a useful framework for further studies aimed at reconstructing forebrain development in vitro for basic research or therapy.

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“…Although the effect of growth factors and morphogens on ES neural patterning can be anticipated by their known role in vivo, the use of ES cultures for studies aimed to steer their neural patterning toward distinct fates, or to investigate in vitro the signaling controlling neuronal fate acquisition, is complicated by the possible production of growth factors and morphogens inside the neuralizing culture. Many studies reported that natural or chemical inhibitors/ antagonists of Wnts (Bouhon et al, 2006;Kirkeby et al, 2012), BMPs (Surmacz et al, 2012;Lupo et al, 2013;Ozair et al, 2013), FGFs (Chambers et al, 2009;Kriks et al, 2011;Lupo et al, 2013), Activin/Nodal (Lupo et al, 2013), or Shh (Gaspard et al, 2008;Li et al, 2009;Nicoleau et al, 2013) affect the differentiation fate of neurons cultured in chemically defined media. These studies implicate that such factors were endogenously produced by ES differentiating cells.…”

Section: Introductionmentioning

confidence: 99%

The double inhibition of endogenously produced BMP and Wnt factors synergistically triggers dorsal telencephalic differentiation of mouse ES cells

Bertacchi

Pandolfini

D’Onofrio

et al. 2014

Developmental Neurobiology

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Embryonic stem (ES) cells are becoming a popular model of in vitro neurogenesis, as they display intrinsic capability to generate neural progenitors that undergo the known steps of in vivo neural development. These include the acquisition of distinct regional fates, which depend on growth factors and signals that are present in the culture medium. The control of the intracellular signaling that is active at different steps of ES cell neuralization, even when cells are cultured in chemically defined medium, is complicated by the endogenous production of growth factors. However, this endogenous production has been poorly investigated so far. To address this point, we performed a high-throughput analysis of the expression of morphogens during mouse ES cell neuralization in minimal medium. We found that during their neuralization, ES cells increased the expression of members of Wnt, Fibroblast Growth Factor (FGF), and BMP families. Conversely, the expression of Activin/Nodal and Shh ligands was low in early steps of neuralization. In this experimental condition, neural progenitors and neurons generated by ES cells expressed a gene expression profile that was consistent with a midbrain identity. We found that endogenous BMP and Wnt signaling, but not FGF signaling, synergistically affected ES cell neural patterning, by turning off a profile of dorsal/telencephalic gene expression. Double BMP and Wnt inhibition allowed neuralized ES cells to sequentially activate key genes of cortical differentiation. Our findings are consistent with a novel synergistic effect of Wnt and BMP endogenous signaling of ES cells in inhibiting a cortical differentiation program.

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Multiple roles of Activin/Nodal, bone morphogenetic protein, fibroblast growth factor and Wnt/β-catenin signalling in the anterior neural patterning of adherent human embryonic stem cell cultures

Cited by 34 publications

References 57 publications

WNT/β-catenin signaling mediates human neural crest induction via a pre-neural border intermediate

WNT/β-catenin signaling mediates human neural crest induction via a pre-neural border intermediate

From pluripotency to forebrain patterning: an in vitro journey astride embryonic stem cells

The double inhibition of endogenously produced BMP and Wnt factors synergistically triggers dorsal telencephalic differentiation of mouse ES cells

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