CORTECON: A Temporal Transcriptome Analysis of In Vitro Human Cerebral Cortex Development from Human Embryonic Stem Cells

Leemput, Joyce van de; Boles, Nathan C.; Kiehl, Thomas R.; Corneo, Barbara; Lederman, Patty; Menon, Vilas; Lee, Changkyu; Martinez, Refugio A.; Levi, Boaz P.; Thompson, Carol L.; Yao, Shuyuan; Kaykas, Ajamete; Temple, Sally; Fasano, Christopher A.

doi:10.1016/j.neuron.2014.05.013

Cited by 181 publications

(208 citation statements)

References 63 publications

(78 reference statements)

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“…Piezo1 also is identified in public gene-expression databases from human embryonic stem (hES) cells (45), mesenchymal stem cells (46), and neural stem cells (47,48). Of particular interest, van de Leemput et al (48) differentiated hES cells into brain organoids to study changes in gene expression accompanying human corticogenesis.…”

Section: Discussionmentioning

confidence: 99%

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Stretch-activated ion channel Piezo1 directs lineage choice in human neural stem cells

Pathak¹,

Nourse

Tran³

et al. 2014

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

483

509

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Neural stem cells are multipotent cells with the ability to differentiate into neurons, astrocytes, and oligodendrocytes. Lineage specification is strongly sensitive to the mechanical properties of the cellular environment. However, molecular pathways transducing matrix mechanical cues to intracellular signaling pathways linked to lineage specification remain unclear. We found that the mechanically gated ion channel Piezo1 is expressed by brainderived human neural stem/progenitor cells and is responsible for a mechanically induced ionic current. Piezo1 activity triggered by traction forces elicited influx of Ca 2+ , a known modulator of differentiation, in a substrate-stiffness-dependent manner. Inhibition of channel activity by the pharmacological inhibitor GsMTx-4 or by siRNA-mediated Piezo1 knockdown suppressed neurogenesis and enhanced astrogenesis. Piezo1 knockdown also reduced the nuclear localization of the mechanoreactive transcriptional coactivator Yes-associated protein. We propose that the mechanically gated ion channel Piezo1 is an important determinant of mechanosensitive lineage choice in neural stem cells and may play similar roles in other multipotent stem cells. (5) and that the mechanical properties of the culturing environment before transplantation can influence the outcome of in vivo stem cell transplants (6). Hence, a molecular and mechanistic understanding of how stem cells process mechanical cues and how this processing results in downstream signaling events and ultimately in fate decisions is needed for greater control over the fate of transplanted cells.Studies in mesenchymal and neural stem cells have revealed the involvement of focal adhesion zones and cytoskeletal proteins, such as integrins, nonmuscle myosin II (7), Rho GTPases (8-10), and vinculin (11), that participate in the generation of cellular traction forces. Recent work also has identified the nucleoskeletal protein lamin-A (12) and the transcriptional coactivators Yap (Yesassociated protein) and Taz (transcriptional coactivator with PDZbinding motif) (13) in mechanotransduction in mesenchymal stem cells. However, the mechanisms by which mechanical cues detected by cellular traction forces are transduced to downstream intracellular pathways of differentiation remain unclear.Ion channels are involved, directly or indirectly, in the transduction of all forms of physical stimuli-including sound, light, temperature, mechanical force, and even gravity-into intracellular signaling pathways. Hence, we wondered whether ion channels could be involved in transducing matrix mechanical cues to intracellular signaling pathways linked to lineage specification. In particular, we focused here on cationic stretch-activated channels (SACs) because they are known to detect mechanical forces with high sensitivity and broad dynamic range and because they are permeable to Ca 2+ , an important second messenger implicated in cell fate (14,15). We examined the role of SACs in neural stem cells, for which mechanical cues influence specification alo...

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Section: Discussionmentioning

confidence: 99%

“…Of particular interest, van de Leemput et al (48) differentiated hES cells into brain organoids to study changes in gene expression accompanying human corticogenesis. Their RNA-seq database (cortecon.…”

Section: Discussionmentioning

confidence: 99%

Stretch-activated ion channel Piezo1 directs lineage choice in human neural stem cells

Pathak¹,

Nourse

Tran³

et al. 2014

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

483

509

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“…The CORTECON system is generated by analyzing RNA-Seq data with nine samples from day 0 to day 77 of the differentiating hESCs (20). PCC assays revealed that the CORTECON system could not reveal the conversion from the pluripotent epiblast to the neural lineage, which progresses quickly during the early development of the human nervous system (Fig.…”

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

confidence: 99%

“…Given that intrinsic regulators, especially TFs, play essential roles in the neural commitment (20,33,34), we sought to identify key TF genes in each of the five modules. By overlapping the stagespecific genes with TF database (35) (www.transcriptionfactor.…”

Section: Regulation Network Of Transcription Factor Genes During Hescmentioning

confidence: 99%

“…Global gene expression profiling technologies such as microarray and RNA sequencing (RNA-Seq) enable highly sensitive analysis of transcriptome associated with the neurodevelopment of hESCs. By using RNA-Seq data with nine samples from days 0 to 77 of the differentiating hESCs, van de Leemput et al (20) establish a COR-TECON system to study human cerebral cortex development in vitro. Meanwhile, single-cell RNA-Seq is applied to identify various classes of neural progenitors (NPCs) and neurons, which is helpful for mapping the early human brain cells (21).…”

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confidence: 99%

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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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Engineered Perineural Vascular Plexus for Modeling Developmental Toxicity

Kaushik

Gupta

Harms

et al. 2020

Adv Healthcare Materials

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There is a vital need to develop in vitro models of the developing human brain to recapitulate the biological effects that toxic compounds have on the brain. To model perineural vascular plexus (PNVP) in vitro, which is a key stage in embryonic development, human embryonic stem cells (hESC)‐derived endothelial cells (ECs), neural progenitor cells, and microglia (MG) with primary pericytes (PCs) in synthetic hydrogels in a custom‐designed microfluidics device are cocultured. The formation of a vascular plexus that includes networks of ECs (CD31+, VE‐cadherin+), MG (IBA1+), and PCs (PDGFRβ+), and an overlying neuronal layer that includes differentiated neuronal cells (βIII Tubulin+, GFAP+) and radial glia (Nestin+, Notch2NL+), are characterized. Increased brain‐derived neurotrophic factor secretion and differential metabolite secretion by the vascular plexus and the neuronal cells over time are consistent with PNVP functionality. Multiple concentrations of developmental toxicants (teratogens, microglial disruptor, and vascular network disruptors) significantly reduce the migration of ECs and MG toward the neuronal layer, inhibit formation of the vascular network, and decrease vascular endothelial growth factor A (VEGFA) secretion. By quantifying 3D cell migration, metabolic activity, vascular network disruption, and cytotoxicity, the PNVP model may be a useful tool to make physiologically relevant predictions of developmental toxicity.

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CORTECON: A Temporal Transcriptome Analysis of In Vitro Human Cerebral Cortex Development from Human Embryonic Stem Cells

Cited by 181 publications

References 63 publications

Stretch-activated ion channel Piezo1 directs lineage choice in human neural stem cells

Stretch-activated ion channel Piezo1 directs lineage choice in human neural stem cells

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

Engineered Perineural Vascular Plexus for Modeling Developmental Toxicity

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