Rational derivation of midbrain dopamine neuron progenitors from hPSCs using a selective FZD5 antibody agonist
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ABSTRACT:
PROVIDER: PRJNA1006285 | ENA |
REPOSITORIES: ENA
Dataset's files
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| SRR25661600.fastq.gz | Fastqsanger.gz | |||
| SRR25661601.fastq.gz | Fastqsanger.gz | |||
| SRR25661602.fastq.gz | Fastqsanger.gz | |||
| SRR25662151.fastq.gz | Fastqsanger.gz | |||
| SRR25662152.fastq.gz | Fastqsanger.gz |
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Project description:Rational derivation of midbrain dopamine neuron progenitors from hPSCs using a selective FZD5 antibody agonist [scRNA-Seq]
| PRJNA1006289 | ENA
Project description:Rational derivation of midbrain dopamine neuron progenitors from hPSCs using a selective FZD5 antibody agonist [RNA-Seq]
| PRJNA1006288 | ENA
Project description:Current ventral midbrain dopaminergic progenitor differentiation protocols utilize small molecule inhibitors targeting Glycogen synthase kinase-3 (GSK3) to activate Wnt signaling, a step required for the anterior-posterior patterning of the nervous system and acquisition of the midbrain fate. However, GSK3a and GSK3β are pleiotropic kinases involved in multiple signaling pathways and their inhibition is a known trigger of neurogenesis. We predicted that direct activation of specific Wnt receptors naturally involved during neural patterning would allow for a more precise spatiotemporal control of Wnt/β-catenin signaling and mimic endogenous cell-cell communication mechanisms to improve hPSC differentiation. Here, we characterized the expression of FZD receptors at the surface of neural progenitor cells with different regional identity. Our data shows that FZD5 expression is uniquely upregulated in anterior neural progenitors and is rapidly downregulated as cells adopt a posterior fate. This spatial regulation of Frizzled cell surface expression constitutes a novel regulatory mechanism adjusting the levels of β-catenin signaling along the anterior-posterior axis and possibly contribute to midbrain-hindbrain boundary formation. Using a tetravalent antibody that selectively triggers FZD5 and LRP6 clustering to activate Wnt/β-catenin signaling, we show that the resulting midbrain-patterned neural progenitors exhibit a gene expression program more closely aligned with the anatomical origin of dopaminergic neurons and could hence represent a more efficient source for cell transplant therapies.
2024-02-21 | GSE241042 | GEO
Project description:Current ventral midbrain dopaminergic progenitor differentiation protocols utilize small molecule inhibitors targeting Glycogen synthase kinase-3 (GSK3) to activate Wnt signaling, a step required for the anterior-posterior patterning of the nervous system and acquisition of the midbrain fate. However, GSK3a and GSK3β are pleiotropic kinases involved in multiple signaling pathways and their inhibition is a known trigger of neurogenesis. We predicted that direct activation of specific Wnt receptors naturally involved during neural patterning would allow for a more precise spatiotemporal control of Wnt/β-catenin signaling and mimic endogenous cell-cell communication mechanisms to improve hPSC differentiation. Here, we characterized the expression of FZD receptors at the surface of neural progenitor cells with different regional identity. Our data shows that FZD5 expression is uniquely upregulated in anterior neural progenitors and is rapidly downregulated as cells adopt a posterior fate. This spatial regulation of Frizzled cell surface expression constitutes a novel regulatory mechanism adjusting the levels of β-catenin signaling along the anterior-posterior axis and possibly contribute to midbrain-hindbrain boundary formation. Using a tetravalent antibody that selectively triggers FZD5 and LRP6 clustering to activate Wnt/β-catenin signaling, we show that the resulting midbrain-patterned neural progenitors exhibit a gene expression program more closely aligned with the anatomical origin of dopaminergic neurons and could hence represent a more efficient source for cell transplant therapies.
2024-02-21 | GSE241041 | GEO
Project description:This SuperSeries is composed of the SubSeries listed below.
2024-02-21 | GSE241043 | GEO
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.
2020-12-18 | GSE162883 | GEO
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
Project description:Naïve human pluripotent stem cells (hPSCs) provide a unique experimental platform of cell fate decisions during pre-implantation development, but their lineage potential remains incompletely characterized. As naïve hPSCs share transcriptional and epigenomic signatures with trophoblast cells, it has been proposed that the naïve state may have enhanced predisposition for differentiation along this extraembryonic lineage. Here we examined the trophoblast potential of isogenic naïve and primed hPSCs. We found that naïve hPSCs can directly give rise to human trophoblast stem cells (hTSCs) and undergo further differentiation into both extravillous and syncytiotrophoblast. In contrast, primed hPSCs do not support hTSC derivation, but give rise to non-self-renewing cytotrophoblasts in response to BMP4. Global transcriptome and chromatin accessibility analyses indicate that hTSCs derived from naïve hPSCs acquire features of pre-implantation trophectoderm. The derivation of hTSCs from naïve hPSCs will enable elucidation of early mechanisms that govern normal human trophoblast development and associated pathologies.
2020-02-13 | GSE138761 | GEO
Project description:Naïve human pluripotent stem cells (hPSCs) provide a unique experimental platform of cell fate decisions during pre-implantation development, but their lineage potential remains incompletely characterized. As naïve hPSCs share transcriptional and epigenomic signatures with trophoblast cells, it has been proposed that the naïve state may have enhanced predisposition for differentiation along this extraembryonic lineage. Here we examined the trophoblast potential of isogenic naïve and primed hPSCs. We found that naïve hPSCs can directly give rise to human trophoblast stem cells (hTSCs) and undergo further differentiation into both extravillous and syncytiotrophoblast. In contrast, primed hPSCs do not support hTSC derivation, but give rise to non-self-renewing cytotrophoblasts in response to BMP4. Global transcriptome and chromatin accessibility analyses indicate that hTSCs derived from naïve hPSCs acquire features of pre-implantation trophectoderm. The derivation of hTSCs from naïve hPSCs will enable elucidation of early mechanisms that govern normal human trophoblast development and associated pathologies.
2020-02-13 | GSE138688 | GEO
Project description:Human pluripotent stem cells (hPSCs) are a promising source of cells for applications in regenerative medicine. Directed differentiation of hPSCs into specialized cells such as spinal motoneurons or midbrain dopamine (DA) neurons has been achieved. However the effective use of hPSCs for cell therapy has lagged far behind. While mouse PSC-derived DA neurons have shown efficacy in models of Parkinson’s disease, DA neurons derived from human PSCs generally display poor in vivo performance. There are also considerable safety concerns for hPSCs related to their potential for teratoma formation or neural overgrowth. Here we present a novel floor plate-based strategy for the derivation of human DA neurons that efficiently engraft, suggesting that past failures were due to incomplete specification rather than a specific vulnerability of the cells. Midbrain floor plate precursors are derived from hPSCs in days following exposure to small molecule activators of sonic hedgehog (SHH) and canonical WNT signaling. Engraftable midbrain DA neurons are obtained by day 25 and can be maintained in vitro for several months. Extensive in vitro molecular profiling, biochemical and electrophysiological data define developmental progression and confirm identity of hPSC-derived midbrain DA neurons. In vivo survival and function is demonstrated in PD animal models in three host species. Long-term engraftment in 6-OHDA-lesioned mouse and rats demonstrates robust survival of midbrain DA neurons, complete restoration of amphetamine-induced rotation behavior and improvements in tests of forelimb use and akinesia. Finally, scalability is demonstrated by transplantation into Parkinsonian monkeys. Excellent DA neuron survival, function and lack of neural overgrowth in the three animal models tested indicate considerable promise for the development of cell based therapies in PD. Differentiated hESC with three conditions (LSB, LSB/S/F8, LSB/S/F8/CHIR) were subjected to RNA extraction in specific timepoint (day 0, 1, 3, 5, 7, 11, 13, 25) and hybridization on Illumina microarrays. Each sample has 3 or 4 biological repeats. Based on previous study* of dual SMAD inhibition neural induction, we developed new midbrain dopamine neuron protocol. It depends on time specific treatment of below factors (LSB/S/F8/CHIR): L (LDN193189 (BMP inhibitor) , day 0-11), SB (SB431542 (TGF-b signal inhibitor), day 0-5), S (SHH + Purmorphamine (Smo agonist), day 1-7), F8 (FGF8, day 1-7) and CHIR (CHIR99021 (GSK3b inhibitor), day 3-13) LSB and LSB/S/F8 are limited control conditions of dual SMAD only (LSB) or traditional patterning with Sonic and FGF (LSB/S/F8) *Chambers,S.M. et al. Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling. Nat. Biotechnol. 27, 275-280 (2009).
2011-11-07 | E-GEOD-32658 | biostudies-arrayexpress