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Homo sapiens

ABSTRACT: Pluripotent stem cell disease modeling of GATA6 haploinsufficiency in human pancreatic development

PROVIDER: PRJNA358077 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR5120780_1.fastq.gz	Fastqsanger.gz
	SRR5120780_2.fastq.gz	Fastqsanger.gz
	SRR5120781_1.fastq.gz	Fastqsanger.gz
	SRR5120781_2.fastq.gz	Fastqsanger.gz
	SRR5120774_1.fastq.gz	Fastqsanger.gz

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Project description:Pluripotent stem cell disease modeling of GATA6 haploinsufficiency in human pancreatic development

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Pluripotent stem cell disease modeling of GATA6 haploinsufficiency in human pancreatic development

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GATA6 regulates WNT and BMP programs to pattern precardiac mesoderm during the earliest stages of human cardiogenesis (RNA-Seq)

Project description:Haploinsufficiency for GATA6 is associated with congenital heart disease (CHD) with variable comorbidity of pancreatic or diaphragm defects, although the etiology of disease is not well understood. Here, we used cardiac directed differentiation from human embryonic stem cells (hESCs) as a platform to study GATA6 function during early cardiogenesis. GATA6 loss-of-function hESCs had a profound impairment in cardiac progenitor cell (CPC) specification and cardiomyocyte (CM) generation due to early defects during the mesendoderm and lateral mesoderm patterning stages. Profiling by RNA-seq and CUT&RUN identified genes of the WNT and BMP programs regulated by GATA6 during early mesoderm patterning. Furthermore, interactome analysis detected GATA6 binding with developmental transcription factors and chromatin remodelers suggesting cooperative regulation of cardiac lineage gene accessibility. We show that modulating WNT and BMP inputs during the first 48 hours of cardiac differentiation is sufficient to partially rescue CPC and CM defects in GATA6 heterozygous and homozygous mutant hESCs. This study provides evidence of the regulatory functions for GATA6 directing human precardiac mesoderm patterning during the earliest stages of cardiogenesis to further our understanding of haploinsufficiency causing CHD and the co-occurrence of cardiac and other organ defects caused by human GATA6 mutations.

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Project description:To better understand molecular mechanisms of human pancreatic development associated with GATA6 haploinsufficiency, we performed RNA-seq analysis to characterize the biological processes affected by GATA6 heterozygous mutation. We identified 542 differential expression genes in the mutants, of which 310 were downregulated. Gene ontology analysis showed that top downregulated transcription factors were associated with developmental process.

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Project description:The mouse is often the model of choice for genetic analysis of neurological disorders, but the introduction of disease mutations into a particular inbred strain sometimes fails to yield phenotypes relevant to human disease. Interrogating genetically diverse panels of mice can identify better models of human sensitivity and resistance to candidate disease variants. We developed an in vitro methodology for modeling multiple stages of central nervous system development using a panel of genetically diverse mouse embryonic stem cell lines. Chemical knockdown of the neurodevelopmental gene Dyrk1a demonstrated profound strain differences in the cellular response to the ablation of DYRK1A activity throughout development in vitro. Responsive strains showed in vitro developmental defects consistent with observations in vivo on Dyrk1a knockout mice, and transcriptomic analysis of sensitive and resistant cell strain backgrounds successfully identified key molecular pathways in neural development known to be associated with Dyrk1a haploinsufficiency in vivo. Thus we demonstrate that high throughput comparative phenotype analysis of differentiated cells from human and genetically diverse mouse pluripotent stem cells bearing disease mutations can provide a facile route to identification of optimal mouse strains for precision disease modeling in vivo.

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