Next generation sequencing of chicken otocyst with pharmacologically induced loss-of-Wnt signalling
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA627935 | ENA |
REPOSITORIES: ENA
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Project description:Bulk RNA-sequencing of individual E2.5 chicken otocyst incubated in media enriched with Wnt signalling inhibitor IWR-1 or control condition (DMSO) for 24h. Transcripts abundance was mapped to a chicken reference genome using Kallisto package and differentially expressed genes were identify using Sleuth package (genes with p-value < 0.05 were considered as significant). Biological Function enrichment analysis (Toppgene online tool) showed that many of these genes were associated with neurogenesis.
2021-04-01 | GSE161638 | GEO
Project description:Bulk RNA-sequencing of individual E2.5 chicken otocyst incubated in media enriched with Wnt signalling inhibitor IWR-1 or control condition (DMSO) for 24h. Transcripts abundance was mapped to a chicken reference genome using Kallisto package and differentially expressed genes were identify using Sleuth package (genes with p-value < 0.05 were considered as significant). The expression of 1340 genes was significantly changed:733 genes were up-regulated and 607 were down-regulated. Biological Function enrichment analysis (Toppgene online tool) showed that many of these genes were associated with neurogenesis.
2020-12-01 | GSE149310 | GEO
Project description:Bulk RNA-sequencing of individual E3 chicken otocyst, in which Wnt signaling activity was reduced by genetic manipulation. The E2 otic placodes were in ovo transfected with a construct overexpressing truncated form of B-catenin reducing Wnt activity or a control plasmid expressing mCherry and collected 24h later. Transcripts abundance was mapped to a chicken reference genome using Kallisto package and differentially expressed genes were identify using Sleuth package (genes with p-value < 0.05 were considered as significant).
2023-11-13 | GSE230083 | GEO
Project description:Next generation sequencing of chicken otocyst with pharmacologicaly induced loss-of-Wnt signalling
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Project description:Bulk RNA-sequencing of individual chicken otocysts, in which Notch activity was either stimulated or blocked. To stimulate Notch activity, the E2 otic placodes were in ovo transfected with a construct overexpressing Notch1 Intracellular domain (NICD1) or a control plasmid expressing mRFP1 and collected 6h or 24h later. To pharmacologically block Notch activity, E2.5 chicken otocysts were incubated in media enriched with γ-secretase Inhibitor or control condition (DMSO) for 6h or 24h. Transcripts abundance was mapped to a chicken reference genome using Kallisto package.
2022-12-01 | GSE196999 | GEO
Project description:Hypothesis and proof of concept:
Our hypothesis is that increase in cellular iron import proteins (TfR1, DMT1) occur early in the adenoma-carcinoma sequence through mutations in APC and lead to cellular iron loading. As demonstrated in our previous work the effects of this iron loading is to mediate increased Wnt signalling resulting in c-myc induction. This in turn serves to increase the expression of iron import proteins (TfR1, DMT1) and decrease the expression of iron export (ferroportin [FPN]) and storage (ferritin) proteins. Such a hypothesis explains how Wnt signalling controls iron metabolism and ensures that there is adequate cellular iron for ATP generation and cellular proliferation.
Experimental design:
To test such a hypothesis we aim to prospectively collect the following colorectal tissue from patients attending for colonoscopy:
1. Normal colonic mucosa in patients with no colorectal pathology (n = 30)
2. Polyps and matched normal colon (n = 30)
3. Colorectal cancers and matched normal colon (n = 30)
We also intend to collect serum and urine from the following patient groups:
4. Normal colonoscopy (n = 30)
5. Colorectal adenomas (n = 30)
6. Colorectal cancers (n = 30)
Primary outcome(s): Measured at baseline, using the expression of proteins in the tissue and serum to detect the cellular and systemic iron transport proteins. The techniques used will include mass spectrometry, western blotting, real time PCR and immunohistochemistry.
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Project description:WNT signalling activity on epiblast cells
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Project description:The function of the FAM83F protein, like the functions of many members of the FAM83 family, is poorly understood. Here we show that injection of Fam83f mRNA into Xenopus embryos causes axis duplication, a phenotype indicative of enhanced Wnt signalling. Consistent with this, overexpression of FAM83F activates Wnt signalling, whilst ablation of FAM83F from human colorectal cancer (CRC) cells attenuates it. We demonstrate that FAM83F is farnesylated and interacts and co-localises with CK1α at the plasma membrane. This interaction with CK1α is essential for FAM83F to activate Wnt signalling, and FAM83F mutants that do not interact with CK1α fail to induce axis duplication in Xenopus embryos and to activate Wnt signalling in cells. FAM83F acts upstream of GSK-3β, because the attenuation of Wnt signalling caused by loss of FAM83F can be rescued by GSK-3 inhibition. Introduction of a farnesyl-deficient mutant of FAM83F in cells through CRISPR/Cas9 genome editing redirects the FAM83F-CK1α complex away from the plasma membrane and significantly attenuates Wnt signalling, indicating that FAM83F exerts its effects on Wnt signalling at the plasma membrane.
2020-12-23 | PXD023121 | Pride
Project description:Ciliary associated extracellular vesicles modulate WNT signalling
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Project description:Primary cilia are microtubule based sensory organelles important for receiving and processing cellular signals. Recent studies have shown that cilia also release extracellular vesicles (EVs) but little is known about their composition and function. Because EVs have been shown to exert various functions in physiology and pathology, these findings have the potential to fundamentally alter our understanding of how the primary cilium is able to regulate specific signalling pathways in development and disease. Compared to control, ciliary mutant mammalian cells demonstrated increased secretion of small EVs (smEVs) and a change in EV composition. Small RNA sequencing and mass spectrometry identified an enrichment for WNT signalling-associated miRNAs and proteins loaded into ciliary mutant EVs. Furthermore, we show that smEVs secreted from mammalian cells are biologically active and modulate the WNT response in recipient cells. These results highlight a possible new smEV-dependent ciliary signalling mechanism which could provide us with new insights into paracrine ciliary signalling as well as ciliopathy disease pathogenesis.
2021-07-19 | PXD020466 | Pride