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Selective activation of FZD7 in human pluripotent stem cells

ABSTRACT: Selective activation of FZD7 in human pluripotent stem cells

PROVIDER: PRJNA664040 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR12659730_1.fastq.gz	Fastqsanger.gz
	SRR12659730_2.fastq.gz	Fastqsanger.gz
	SRR12659731_1.fastq.gz	Fastqsanger.gz
	SRR12659731_2.fastq.gz	Fastqsanger.gz
	SRR12659732_1.fastq.gz	Fastqsanger.gz

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Selective activation of FZD7 in human pluripotent stem cells

Project description:Human pluripotent stem cells ( (H1/WA01/NIH stem cell registry number 0043) were treated with WNT3a protein, a FZD7-specific WNT mimetic, and a GSK3 inihibitor in time series to determine genes differentially upregulated and downregulated across all conditions or within one condition only.

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Selective activation of FZD2 and FZD7 reveals non-redundant function during mesoderm differentiation

Project description:Wnt-ßcatenin signalling controls stem cell self-renewal and cell fate decisions and therefore its precise control could influence differentiation protocols efficiency and cell therapy efforts. During gastrulation, Wnt-ßcatenin signalling dictates lineage bifurcation choices leading from pluripotency to distinct primitive streak patterning, ultimately resulting in various mesendoderm cell types. However, the nature of the Wnt receptors involved in this process and how their signaling dynamics impact mesoderm specification remain elusive. Here, we used selective Frizzled and LRP5/6 antibody-based agonists (FLAgs) to investigate the function and activation kinetics of Frizzled receptor complexes during directed mesoderm differentiation of hPSCs. We found that FZD2 and FZD7 are expressed at the surface of hPSCs and that their activation triggers ßcatenin signaling with different kinetics thereby influencing mesoderm patterning choices. Using bulk and single-cell RNA sequencing, we showed that FZD7 activation promotes lineage commitment towards both paraxial and lateral mesoderm whereas FZD2 engagement preferentially induces paraxial mesoderm. Mechanistically, our results demonstrate that although the short-term response following FZD2 and FZD7 activation is similar, ßcatenin signaling kinetics differs. FZD2 activation results in a sustained Wnt activation profile that drives hPSCs differentiation to paraxial mesoderm while blocking lateral mesoderm. In contrast, FZD7 activation kinetics consists of a similar initial activation followed by dampening of the response to a low level of ßcatenin signaling that is permissive for lateral mesoderm induction in addition to paraxial mesoderm specification. Our results suggest that the function of FZD2 and FZD7 during mesoderm specification is non-redundant and that their selective activation to temporally modulate Wnt-ßcatenin signaling can be leveraged for precise directed differentiation outcomes.

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Metabolomics Workbench ST000003 - GNPS - Metabolomic analysis of mouse embryonic fibroblasts, embryonic stem cells, and induced pluripotent stem cells

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Selective activation of FZD2 and FZD7 reveals non-redundant function during mesoderm differentiation

Project description:Selective activation of FZD2 and FZD7 reveals non-redundant function during mesoderm differentiation

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Expression data from human pluripotent stem cells and differentiated cells treated with pluripotent-specific inhibitors

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Project description:Conventional reprogramming methods rely on the ectopic expression of transcription factors to reprogram somatic cells into induced pluripotent stem cells (iPSC). Forced expression of transcription factors might lead to off-target gene activation and heterogeneous reprogramming, resulting in the emergence of alternative cell types and aberrant iPSC. Activation of endogenous pluripotency factors by CRISPR activation (CRISPRa) can potentially reduce this heterogeneity. Here we describe a high efficiency reprogramming of human somatic cells into iPSC using optimized CRISPRa. Efficient reprogramming was dependent on the additional targeting of the embryo genome activation-enriched Alu-motif and the miR-302/367 locus. Single-cell transcriptome analysis revealed that the optimized CRISPRa reprogrammed cells with more directly and specifically into the pluripotent state when compared to the conventional reprogramming method. These findings support the use of CRISPRa for high quality reprogramming of human iPSC. This SuperSeries is composed of the SubSeries listed below.

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