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Mycn regulates intestinal development through ribosomal biogenesis in a zebrafish model of Feingold syndrome 1 [single-cell RNA-seq]

ABSTRACT: Purpose: To systematically investigate the phenotypes resulting from Mycn loss-of-function at a higher resolution. Methods: Approximately 30 mycn mutant or WT zebrafish embryos at 72 hpf (hours post fertilization) were harvested . Libraries were prepared using Chromium Controller and Chromium Single Cell 3’Library & Gel Bead Kit v3 (10x Genomics, PN-1000074) according to the manufacturer’s protocol for 10000 cells recovery. Results: A total of 25,886 single-cell transcriptomes were collected after stringent quality control measures. Conclusions: 27 Cell types were identified by intergrating two datasets and 9 subclusters were obtained by reclustering intestinal cells.

ORGANISM(S): Danio rerio

PROVIDER: GSE191002 | GEO | 2021/12/17

REPOSITORIES: GEO

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Mycn regulates intestinal development through ribosomal biogenesis in a zebrafish model of Feingold syndrome 1 (RNA-seq and Ribo-seq)

Project description:Purpose: To systematically investigate the molecular mechanisms resulting from Mycn loss-of-function. Methods: Approximately 30 mycn mutant or WT zebrafish embryos at 72 hpf (hours post fertilization) were harvested . Libraries were prepared using Chromium Controller and Chromium Single Cell 3’Library & Gel Bead Kit v3 (10x Genomics, PN-1000074) according to the manufacturer’s protocol for 10000 cells recovery. Results: More than 3,000 genes showed reduced tranlation efficiency in Mycn mutant. Conclusions: mTOR signaling was reduced in Mycn mutant.

2022-08-26 | GSE211652 | GEO

Nodal coordinates the anterior-posterior patterning of germ layers and induces head formation in zebrafish explants.

Project description:Purpose: To investingate cell types in wild-type zebrafish explants. Methods: Wild-type explants (injected with phenol red) were harvested at 10hpf. Libraries were prepared using Chromium Controller and Chromium Single Cell 3’Library & Gel Bead Kit v3 (10x Genomics, PN-1000075) according to the manufacturer’s protocol for 10000 cells recovery. Results: A total of 9,968 single-cell transcriptomes were collected after stringent quality control measures. Conclusions: 6 Cell types were identified, wild-type explants mainly contains the anterior neural ectoderm and epidermis at 10 hpf.

2022-12-22 | GSE221220 | GEO

Differential Nodal level promotes mesendoderm cell fate segregation mediated by chromatin organization

Project description:Purpose: To investigate the mechanism of prechordal plate and anterior endoderm separation . Methods: Nodal injected explants (injected with 10pg ndr2 mRNA) constructed from lft1 mutants and ndr1 morphants were harvested at 6hpf. Libraries were prepared using Chromium Controller and Chromium Single Cell 3’Library & Gel Bead Kit v3 (10x Genomics, PN-1000075) according to the manufacturer’s protocol for 10000 cells recovery. For single-cell multiomics, zebrafish embryos at 6 hpf were harvested. Libraries were prepared using Chromium Next GEM Single Cell Multiome ATAC + Gene Expression Reagent Bundle (10x Genomics, 4 rxns PN-1000285) according to the manufacturer’s protocol for 10000 cells recovery. Results: A total of 10,614 single-cell transcriptomes and 4,335 multiomics were collected after stringent quality control measures. Conclusions: A slight bias in Nodal signaling promotes a differential chromatin structure between prechordal plate and endoderm, which drives a differential expression of those key regulators, such as gsc and ripply1 in these two cell lineages, and further regulates mesendoderm cell fate separation.

2023-01-28 | GSE223636 | GEO

Single cell response landscape of graded Nodal signaling in zebrafish explants [single-cell RNA-seq]

Project description:Purpose: To construct cell landscape of graded Nodal signaling in zebrafish explants. Methods: Nodal injected explants (injected with 10pg ndr2 mRNA) were harvested at 4hpf, 5hpf, 6hpf, 8hpf, 10hpf, 24hpf. Libraries were prepared using Chromium Controller and Chromium Single Cell 3’Library & Gel Bead Kit v2 (10x Genomics, PN-120237) according to the manufacturer’s protocol for 10000 cells recovery. Results: A total of 42,317 single-cell transcriptomes were collected after stringent quality control measures. Conclusions: 19 Cell types from 6 developmental stages were identified, Nodal explant mainly contains the axial mesoderm (prechordal plate and notochord) and anterior-posterior patterned head structures.

2021-01-28 | GSE165653 | GEO

Mycn regulates intestinal development through ribosomal biogenesis in a zebrafish model of Feingold syndrome 1 [bulk RNA-seq]

Project description:Purpose: To identify differential expressed genes for loss of function of Mycn in zebrafish Methods: wild-type and mycn mutant zebrafish embryos were collected for RNA sequencing at 48 hpf (hours post fertilisation) and 72 hpf Results: We found 85 (overlapped) down-regulated genes in mycn mutant embryos at 48 hpf and 72 hpf (padj < 0.1, log2 fold change < 0) Conclusions: 85 down-regulated genes were identified

2021-12-17 | GSE191001 | GEO

in vitro construction of an embryonic caudal organizer

Project description:Purpose: Characterization of cell types in bmp4 explants and BMP4 induced human embryoids. Methods: Single cell transcriptomes of bmp4 explants were harvested at 6 hpf, 8 hpf,10 hpf, 12 hpf, 18 hpf and 24 hpf. Single cell transcriptomes of BMP4 induced human embryoids were harvested at Day 5 and Day 6. Libraries were prepared using Single Cell 3′Library & gel Bead kit v3.1 (10x Genomics, Cat# PN-1000121) according to the manufacturer’s protocol for 10000 cell recovery. Results: paraxial mesoderm, lateral plate mesoderm, and their derivatives were detected in bmp4 explants, and NMP, somite and neural cell fates were detected in bmp4 induced human embryoids. Conclusions: A well-organized caudal-like structure was induced in bmp4 explants and caudal cell fates were specified in BMP4 induced human embryoids.

2024-07-08 | GSE270989 | GEO

Single-cell RNAseq analysis of H1N1-induced lung injury

Project description:The lungs of mice inoculated with H1N1 were harvested at day 7. The DAPI- lung cells were loaded into a 10x Genomics microfluidics chip and encapsulated with barcoded oligo-dT-containing gel beads using the 10x Genomics Chromium controller using v3 kit according to the manufacturer's instructions.

2022-04-29 | GSE201541 | GEO

ScRNA-seq and scATAC-seq analysis of human pluripotent stem cell (hPSC) derived macrophages iMacs

Project description:Purpose: to depict the single-cell map and reveal the heterogeneity of hPSC derived macrophages Methods: Differentiation day 14 iMacs were dissociated to obtain a single-cell suspension. scRNA-seq and scATAC-seq library preparation and sequencing were done by CapitalBio Technology (https://www.capitalbiotech.com/). scRNA-seq, the cell suspension was immediately loaded onto the Chromium Single Cell Controller (10× Genomics) for droplet formation. cDNA and subsequent sequencing library were generated using Chromiun Single Cell 3’ Reagent Kit (v3 Chemistry) (10× Genomics) according to the instruction manual. For sc-ATAC-seq, cell nuclei were extracted with Chromiun Nuclei Isolation Kit (v2 Chemistry) (10x Genomics) and then loaded onto the Chromium Single Cell Controller for droplet formation. Results: ScRNA-seq and scATAC-seq data of differentiation day 14 iMacs from hPSCs Conclusions: day 14 differentiation culture, which consisted primarily of monocytes and macrophages.

2024-05-23 | GSE234577 | GEO

Neuroblastoma: MYCN normal vs. MYCN amplified

Project description:To evaluate the expression of genes associated with MYCN in NB, 10 tumors with MYCN amplification and 10 with normal MYCN copy number were subjected to oligonucleotide microarray using Agilent oligo microarray chips. Two-condition experiment, MYCN normal vs. MYCN amplified 10 NB patient tissues for each group.

2013-12-17 | E-GEOD-53371 | biostudies-arrayexpress

Metabolic dependencies in MYCN-driven neuroblastoma

Project description:Here we sought metabolic alterations specifically associated with amplified MYCN as nodes to indirectly target the MYCN oncogene. Liquid chromatography-mass spectrometry-based proteomics identified 7 proteins consistently correlated with MYCN in proteomes from 49 neuroblastoma biopsies and 13 cell lines. Among these were phosphoglycerate dehydrogenase (PHGDH), the rate-limiting enzyme in de novo serine synthesis. MYCN associated with two regions in the PHGDH promoter, supporting transcriptional PHGDH regulation by MYCN. Pulsed stable isotope-resolved metabolomics utilizing 13C-glucose labeling demonstrated higher de novo serine synthesis in MYCN-amplified cells compared to cells with diploid MYCN. An independence of MYCN-amplified cells from exogenous serine and glycine was demonstrated by serine and glycine starvation, which attenuated nucleotide pools and proliferation only in cells with diploid MYCN but did not diminish these endpoints in MYCN-amplified cells. Proliferation was attenuated in MYCN-amplified cells by CRISPR/Cas9-mediated PHGDH knockout or treatment with PHGDH small molecule inhibitors without affecting cell viability. PHGDH inhibitors administered as single-agent therapy to NMRI-Foxn1nu/nu mice harboring patient-derived MYCN-amplified neuroblastoma xenografts slowed tumor growth. However, combining a PHGDH inhibitor with the standard-of-care chemotherapy drug, cisplatin, revealed antagonism of chemotherapy efficacy in vivo. Emergence of chemotherapy resistance was confirmed in the genetic PHGDH knockout model in vitro. Altogether, PHDGH knockout and inhibition by small molecules consistently slows proliferation, but stops short of killing the cells, which then establish resistance to classical chemotherapy. Although PHGDH inhibition with small molecules has produced encouraging results in other preclinical cancer models, this approach must be considered with caution in patients with neuroblastoma.

2020-12-11 | PXD018300 | Pride

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