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Gene expression profiling of human neural crest cells and smooth muscle cells treated with DMSO or Nutlin3a

ABSTRACT: Gene expression profiling of human neural crest cells and smooth muscle cells treated with DMSO or Nutlin3a

PROVIDER: PRJNA526696 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR8715400_1.fastq.gz	Fastqsanger.gz
	SRR8715400_2.fastq.gz	Fastqsanger.gz
	SRR8715391_1.fastq.gz	Fastqsanger.gz
	SRR8715391_2.fastq.gz	Fastqsanger.gz
	SRR8715401_1.fastq.gz	Fastqsanger.gz

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Gene expression profiling of human neural crest cells and smooth muscle cells treated with DMSO or Nutlin3a

Project description:Activation of the p53 transcription factor triggers apoptosis only in certain cell types. Human neural crest cells (hNCCs), which can be generated from human embryonic stem cells, undergo apoptosis when treated with the p53 activating drug Nutlin3a. In contrast, when hNCCs are differentiated towards the smooth muscle cell lineage (early-hSMCS), they no longer undergo apoptosis in response to Nutlin3a treatment. Here, we sought to determine whether the transcriptional response to p53 activation differs between cells that are susceptible to p53-driven apoptosis and cells that are resistant to p53-driven apoptosis. To this end, we performed RNA-seq to assess the transcriptional response to Nutlin3a treatment in hNCCs and early-hSMCs.

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A Human Model of Cerebral Amyloid Angiopathy Using Neural Crest-Derived Vascular Smooth Muscle Cells

Project description:There is a growing recognition of cerebrovascular contribution to neurodegenerative diseases. Cerebral amyloid angiopathy (CAA), characterised by amyloid-beta (AM-NM-2) deposits in the walls of intracerebral and leptomeningeal arteries, is evident in a majority of AlzheimerM-bM-^@M-^Ys disease patients and aged people. Here, we leverage on human pluripotent stem cells to generate vascular smooth muscle cells (SMCs) from neural crest progenitors, recapitulating brain vasculature-specific attributes in AM-NM-2 metabolism. We confirm that the lipoprotein receptor, LRP1, functions in our neural crest-derived SMCs to mediate AM-NM-2 uptake and intracellular proteasomal degradation. Hypoxia significantly compromises the ability of SMCs in AM-NM-2 clearance by suppressing LRP1 expression. This enables us to develop an assay of AM-NM-2 uptake using the neural crest-derived SMCs with hypoxia as a stress paradigm. We then tested several vascular protective compounds in a high throughput format, demonstrating the value of stem cell-based phenotypic screening for novel CAA therapeutics and drug repurposing. We adopted our previous SMC differentiation protocol (Cheung et al., 2012) to differentiate this intermediate neural crest population using platelet-derived growth factor BB (PDGF-BB, 10 ng/ml) and transforming growth factor-beta 1 (TGF-M-NM-21, 2ng/ml) for another 12 days. The resultant neural crest-derived SMCs (NCSMC) were then characterised in comparison to neuroectoderm-derived SMCs (NESMC) (Cheung et al., 2012) and positive control, human brain vascular SMCs (BVSMC).

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Human ESC differentiation into cranial neural crest cells in the presence of PhenDC3

Project description:Human ESC differentiation into cranial neural crest cells in the presence of PhenDC4. Human embryonic stem cells (hESC) were differentiated into cranial neural crest cells in presence of the G-quadruplex stabilising ligand PhenDC3 or DMSO control. The case when no PhenDC3 or DMSO have been included in the differentiation protocol represent an additional control experiment.

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Embryonic and adult neural crest stem cells

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Project description:We utilized quantitative analyses of the proteome, transcriptome, and ubiquitinome to study how ubiquitination and NEDD4 control neural crest cell survival and stem-cell-like properties. We report 276 novel NEDD4 targets in neural crest cells and show that loss of NEDD4 leads to a striking global reduction in specific ubiquitin lysine linkages.

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Profiling of microRNA expression of mouse cardiac neural crest cells and non neural crest cells

Project description:Profiling of microRNA expression of mouse cardiac neural crest cells and non neural crest cells

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A Robust Method To Derive Functional Neural Crest Cells From Human Pluripotent Stem Cells

Project description:Neural crest (NC) cells contribute to the development of many complex tissues. The abnormal development of NC cells accounts for a number of congenital birth defects. Generating NC cells, and more specifically NC subpopulations such as cranial, cardiac, and trunk NC cells from human induced pluripotent stem (iPS) cells and human embryonic stem (ES) cells presents a valuable tool to model and study human NC development and disease. Here we provide a robust, efficient, and reproducible protocol for the differentiation of human iPS and ES cells into NC cells. The protocol has been validated in multiple human pluripotent stem cell lines and yields relatively pure NC cell populations in eight days. The resulting cells can be propagated and retain NC marker expression over multiple passages. The NC cells show proper cell specification and can develop into NC-derived cell lineages including smooth muscle cells, peripheral neurons, and Schwann cells. Additionally, the NC cells are functional and migrate to appropriate chemoattractants such as SDF-1, Fgf8b, BMP2, and Wnt3a. Importantly, this method generates all NC subpopulations (cranial, cardiac, and trunk) providing a great advantage to readily available NC differentiation methods. Neural crest cells derived from human induced pluripotent stem cells were profiled using Affymetrix Gene 1.0 arrays to identify differential gene expression changes and alternative exons from the open-source software AltAnalyze. An FDR adjusted emperical Bayes moderated t-test p < 0.05 was used to identify differentially expressed Ensembl genes and GO-Elite used to identify biologically relevant, Ontology, pathway and gene-set categories. Alternative exons were obtained using the FIRMA analysis option and default thresholds. Other array neural crest array and RNA-Seq dataset were compared to this to identify common and distinct regulatory mechanisms.

2013-08-16 | E-GEOD-44727 | biostudies-arrayexpress

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