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Characterization of Wnt-Tcf functions during Ciona heart development

ABSTRACT: Characterization of Wnt-Tcf functions during Ciona heart development

PROVIDER: PRJNA508201 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR8274750.fastq.gz	Fastqsanger.gz
	SRR8274751.fastq.gz	Fastqsanger.gz
	SRR8274752.fastq.gz	Fastqsanger.gz
	SRR8274745.fastq.gz	Fastqsanger.gz
	SRR8274746.fastq.gz	Fastqsanger.gz

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Characterization of Wnt-Tcf functions during Ciona heart development

Project description:To assay Tcf functions during early cardiopharyngeal specification, bulk RNAseq were performed to profile the FACS-purified cardiopharyngeal lineage cells isolated from 15 and 18hpf Ciona larvae with CRISPR/Cas9-mediated Tcf mutagenesis.

2018-12-04 | GSE123295 | GEO

Initial characterization of Wnt-Tcf functions during Ciona heart development.

Project description:In vertebrate embryos, the cardiopharyngeal mesoderm gives rise to both cardiac and branchiomeric head muscles. The canonical Wnt signaling pathway regulates many aspects of cardiomyocyte specification, and modulates a balance between skeletal and cardiac myogenesis during vertebrate head muscle development. However, the role of Wnt signaling during ascidian cardiopharyngeal development remains elusive. Here, we documented the expression of Wnt pathway components during cardiopharyngeal development in Ciona, and generated tools to investigate potential roles for Wnt signaling, and its transcriptional effector Tcf, on heart vs. pharyngeal muscle fate specification. Neither focused functional analyses nor lineage-specific transcriptome profiling uncovered a significant role for Tcf during early cardiac vs. pharyngeal muscle fate choice. By contrast, Wnt gene expression patterns of Frizzled4 and Lrp4/8 and CRISPR/Cas9-mediated Tcf knock-down suggested a later requirement for Wnt signaling during heart morphogenesis and/or cardiomyocyte differentiation. This study provides a provisional set of reagents to study Wnt signaling function in Ciona, and promising insights for future analyses of Wnt functions during heart organogenesis.

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TCF/LEF dependent and independent regulation of Wnt/beta-catenin transcription

Project description:During canonical Wnt signalling the activity of nuclear beta-catenin is largely mediated by the TCF/LEF family of transcription factors. To challenge this view we used the CRISPR/Cas9 genome editing approach to generate HEK 293T cell clones simultaneously carrying loss-of-function alleles of all four TCF/LEF genes. Exploiting unbiased whole transcriptome sequencing studies, we found that a subset of beta-catenin transcriptional targets did not require TCF/LEF factors for their regulation. Consistent with this finding, we observed in a genome-wide analysis that beta-catenin occupied specific genomic regions in the absence of TCF/LEF. Finally, we revealed the existence of a transcriptional activity of beta-catenin that specifically appears when TCF/LEF factors are absent, and refer to this as beta-catenin-GHOST response. Collectively, this study uncovers a previously neglected modus operandi of beta-catenin that bypasses the TCF/LEF transcription factors.

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TCF/LEF dependent and independent regulation of Wnt/beta-catenin transcription

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Long-lasting WNT-TCF response blocking and epigenetic modifying activities of withanolide F in human cancer cells

Project description:The WNT-TCF signaling pathway participates in adult tissue homeostasis and repair, and is hyperactive in a number of human diseases including cancers of the colon. Whereas to date there are no antagonists approved for patient use, a potential problem for their sustained use is the blockade of WNT signaling in healthy tissues, thus provoking potentially serious co-lateral damage. Here we have screened a library of plant and microorganism small molecules for novel WNT signaling antagonists and describe withanolide F as a potent WNT-TCF response blocker. This steroidal lactone inhibits TCF-dependent colon cancer xenograft growth and mimics the effects of genetic blockade of TCF and of ivermectin, a previously reported WNT-TCF blocker. However, withanolide F is unique in that it imposes a long-lasting repression of tumor growth, WNT-TCF targets and cancer stem cell clonogenicity long after drug treatment

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