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Next Generation Sequencing of first trimester human cytotrophoblasts expressing a constitutively active mutant (5SA) of the transcriptional co-activator YAP.

ABSTRACT: Next Generation Sequencing of first trimester human cytotrophoblasts expressing a constitutively active mutant (5SA) of the transcriptional co-activator YAP.

PROVIDER: PRJNA601914 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR10907673.fastq.gz	Fastqsanger.gz
	SRR10907674.fastq.gz	Fastqsanger.gz
	SRR10907675.fastq.gz	Fastqsanger.gz
	SRR10907676.fastq.gz	Fastqsanger.gz
	SRR10907677.fastq.gz	Fastqsanger.gz

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Project description:Invasion of cytotrophoblasts into uterine tissues is essential for placental development. To identify molecules regulating trophoblast invasion, mRNA signatures of purified villous (CTB, poor invasiveness) and extravillous (EVT, high invasiveness) trophoblasts isolated from first trimester human placentae and villous explant cultures, respectively, were compared using GeneChip analyses yielding 991 invasion/migration related transcripts. Several genes involved in physiological and pathologic cell invasion, including ADAM-12,-19,-28 as well as Spondin-2, were upregulated in EVT. Pathway prediction analyses identified several functional modules associated with either the invasive or the non-invasive trophoblast phenotype. One of the genes which were downregulated in the invasive mRNA pool, heme oxygenase-1 (HO-1), was selected for functional analyses. Real-time PCR analyses, Western blottting, and immunofluorescene of first trimester placentae and differentiating villous explant cultures demonstrated downregulation of HO-1 in invasive EVT as compared to CTB. Modulation of HO-1 expression in loss-of as well as gain-of function cell models (BeWo and HTR8/SVneo, respectively) demonstrated an inverse relationship of HO-1 expression with trophoblast migration in transwell and wound healing assays. Importantly, HO-1 expression led to an increase in protein levels and activity of the nuclear hormone receptor PPARgamma. Pharmacological inhibition of PPARgamma abrogated the inhibitory effects of HO-1 on trophoblast migration. Collectively, our results demonstrate that gene expression profiling of EVT and CTB can be used to unravel novel regulators of cell invasion. Accordingly, we identify heme oxygenase-1 as a negative regulator of trophoblast motility acting via upregulation of PPARgamma. Experiment Overall Design: To identify genes potentially regulating cell invasion trophoblast cells of early human gestation with distinct invasive properties were profiled. Experiment Overall Design: Distinct gene expression signatures of highly invasive EVT (n = 6) and poorly invasive CTB (n = 5) of different first trimester placentae using Affymetrix U133A GeneChips interrogating >20,000 genes were determined.

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Next Generation Sequencing of first trimester human cytotrophoblasts expressing a constitutively active mutant (5SA) of the transcriptional co-activator YAP.

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