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Biphasic activation of WNT signaling enhances the derivation of midbrain dopamine neurons for translational use.

ABSTRACT: Biphasic activation of WNT signaling enhances the derivation of midbrain dopamine neurons for translational use.

PROVIDER: PRJNA683629 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR13220400_1.fastq.gz	Fastqsanger.gz
	SRR13220400_2.fastq.gz	Fastqsanger.gz
	SRR13220401_1.fastq.gz	Fastqsanger.gz
	SRR13220401_2.fastq.gz	Fastqsanger.gz
	SRR13220402_1.fastq.gz	Fastqsanger.gz

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Biphasic activation of WNT signaling enhances the derivation of midbrain dopamine neurons for translational use [ChIP-seq]

Project description:We report a novel midbrain dopamine neuron derivation protocol from human pluripotent stem cells (hPSC)s based on a two-step WNT signaling activation strategy. We perform genome-wide chromatin immunoprecipitation (ChIP-seq) for histone modifications including H3K4me3 and H3K27me3 with midbrain dopamine neuron differentiated cells from hPSC in Low-Chir and Boost-Chir (7.5 uM) conditions at day 11. Those results indicate that CHIR-boost promotes a more open chromatin structure at midbrain regions such as an EN1 locus based on increased H3K4me3 and decreased H3K27me3 levels. In contrast, analysis of the contaminating posterior (hindbrain) and anterior (diencephalic) lineage including NKX2-2 and PAX6 loci showed the opposite pattern with decreased H3K4me3 and increased H3K27me3 levels under CHIR-boost conditions. The protocol presented here is the basis for clinical grade dopamine neuron production and preclinical safety and efficacy studies.

2020-12-18 | GSE162882 | GEO

Biphasic activation of WNT signaling enhances the derivation of midbrain dopamine neurons for translational use [RNA-seq]

Project description:We report a novel midbrain dopamine neuron derivation protocol from human pluripotent stem cells (hPSC)s based on a two-step WNT signaling activation strategy. By comparing next-generation sequencing (NGS) derived transcriptome profiling (RNA-seq) of dopamine neuron differentiated cells derived from hPSC with different concentration of Chir-boosting (day4 – day9) measured at day 11 including hPSC, we find that CHIR-boosting improves expression of midbrain markers such as EN1 while minimizing expression of contaminating posterior (hindbrain) and anterior (diencephalic) lineage markers. Furthermore, gene expression analysis from RNA-seq study confirmed the progressive differentiation and maturation of Boost CHIR treated precursors into mDA neurons by day 30 of differentiation. The protocol presented here is the basis for clinical grade dopamine neuron production and preclinical safety and efficacy studies.

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Biphasic activation of WNT signaling enhances the derivation of midbrain dopamine neurons for translational use [ChIP-seq]

Project description:Biphasic activation of WNT signaling enhances the derivation of midbrain dopamine neurons for translational use [ChIP-seq]

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Biphasic activation of WNT signaling enhances the derivation of midbrain dopamine neurons for translational use [RNA-seq]

Project description:Biphasic activation of WNT signaling enhances the derivation of midbrain dopamine neurons for translational use [RNA-seq]

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Biphasic activation of WNT signaling enhances the derivation of midbrain dopamine neurons for translational use.

Project description:This SuperSeries is composed of the SubSeries listed below.

2020-12-18 | GSE162884 | GEO

Biphasic Activation of WNT Signaling Facilitates the Derivation of Midbrain Dopamine Neurons from hESCs for Translational Use.

Project description:Human pluripotent stem cells show considerable promise for applications in regenerative medicine, including the development of cell replacement paradigms for the treatment of Parkinson's disease. Protocols have been developed to generate authentic midbrain dopamine (mDA) neurons capable of reversing dopamine-related deficits in animal models of Parkinson's disease. However, the generation of mDA neurons at clinical scale suitable for human application remains an important challenge. Here, we present an mDA neuron derivation protocol based on a two-step WNT signaling activation strategy that improves expression of midbrain markers, such as Engrailed-1 (EN1), while minimizing expression of contaminating posterior (hindbrain) and anterior (diencephalic) lineage markers. The resulting neurons exhibit molecular, biochemical, and electrophysiological properties of mDA neurons. Cryopreserved mDA neuron precursors can be successfully transplanted into 6-hydroxydopamine (6OHDA) lesioned rats to induce recovery of amphetamine-induced rotation behavior. The protocol presented here is the basis for clinical-grade mDA neuron production and preclinical safety and efficacy studies.

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Chronic hyperactivation of midbrain dopamine neurons causes preferential dopamine neuron degeneration

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