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Comparative analyses of erythroid cells from ES cells and cord blood CD34+ cells reveal mechanisms for defective expansion and enucleation of ES cell-derived erythroid cells

ABSTRACT: Comparative analyses of erythroid cells from ES cells and cord blood CD34+ cells reveal mechanisms for defective expansion and enucleation of ES cell-derived erythroid cells

PROVIDER: PRJNA744932 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR15076230_1.fastq.gz	Fastqsanger.gz
	SRR15076230_2.fastq.gz	Fastqsanger.gz
	SRR15076231_1.fastq.gz	Fastqsanger.gz
	SRR15076231_2.fastq.gz	Fastqsanger.gz
	SRR15076232_1.fastq.gz	Fastqsanger.gz

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Comparative analyses of erythroid cells from ES cells and cord blood CD34+ cells reveal mechanisms for defective expansion and enucleation of ES cell-derived erythroid cells

Project description:We used both RNA-seq and ATAC-seq analyses to uncovered mechanisms for the limited expansion and impaired enucleation in ES CD34+ cell-derived orthochromatic erythroblasts

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Alpha 4 integrin deficiency in human CD34+ cells engenders a precocious erythroid differentiation ,but inhibits enucleation.

Project description:The functional impact of integrin expression in erythropoiesis has been previously emphasized through its decisive influence on erythroid cell-microenvironmental (matrix and cellular) interactions especially under conditions of stress. Beyond that in several in vitro studies the relationship between the two erythroid integrins, a4 and a5, has been incongruous in terms of a proliferative support, either synergistic or antagonistic, whereas a dominant influence of a4 integrin on terminal erythropoiesis in vitro and in vivo has been consistently emphasized. However, the specific cellular and molecular details of this effect have not been defined, especially for human cells. In the present study we have cultured human CD34+ progenitor cells with induced deficiency of a4 integrin (shRNAa4) under erythroid differentiation conditions, in which expanded erythroid progenitor cells are directed to terminal erythroid maturation stages in the absence of any microenvironmental influence. Our data documented that early proliferative expansion in cells lacking a4 expression is significantly limited, but, although erythroid differentiation can proceed normally to terminal stages, their enucleation is drastically impaired. This novel aspect of a4 integrin participation in the enucleation process in vitro resonates on the lack of in vivo enucleation of primitive erythroid cells lacking any integrin expression but affecting adult cells only under stress conditions.

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Effects of miRNA-30a 5p knock down in bone marrow CD71-positive erythroid precursor cells (imBMEP-A)

Project description:Ex vivo production of red blood cells (RBCs) represents a promising alternative for transfusion medicine. Several strategies were described to generate erythroid cell lines from different sources, including embryonic, induced pluripotent and hematopoietic stem cells. With imBMEP-A, we have previously generated the first erythroid cell line directly derived from reticulocyte progenitors. However, the translation of cultured RBCs into clinical use is currently impeded by the inability to scale up to clinically meaningful doses primarily due to impaired in vitro enucleation of RBC progenitor cell lines in general. To identify factors contributing to the inefficient enucleation in immortalized RBC progenitor cell lines, we conducted a proteome analysis of the imBMEP cell line in comparison to hematopoietic stem cells (HSCs). For a detailed description of the cell culture condition, please see linked publication.

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Downregulation of Myc is essential for terminal erythroid maturation

Project description:Terminal differentiation of mammalian erythroid progenitors involves 4-5 cell divisions and induction of many erythroid important genes, followed by chromatin and nuclear condensation and enucleation. The protein levels of c-myc (Myc) are reduced dramatically during late stage erythroid maturation, coinciding with cell cycle arrest in G1-phase and enucleation, suggesting possible roles for c-myc in either or both of these processes. Here we demonstrate that ectopic Myc expression affects terminal erythroid maturation in a dose-dependent manner. Expression of Myc at physiological levels did not affect erythroid differentiation or cell cycle shutdown, but specifically blocked erythroid nuclear condensation and enucleation. Myc prevented deacetylation of several lysine residues in histones H3 and H4 that are normally deacetylated during erythroid maturation. When over-expressed at levels higher than the physiological range, Myc blocked erythroid differentiation and the cells continued to proliferate in cytokine-free, serum-containing culture medium with an early erythroblast morphology. These studies reveal an important dose-dependent function of Myc in regulating terminal maturation in mammalian erythroid cells. Our findings further support the emerging notion that Myc regulates chromatin structure by regulating global histone acetylation states. Five groups with three biological replicates in each.

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Downregulation of Myc is essential for terminal erythroid maturation

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Alpha 4 integrin deficiency in human CD34+ cells engenders a precocious erythroid differentiation ,but inhibits enucleation.

Project description:Alpha 4 integrin deficiency in human CD34+ cells engenders a precocious erythroid differentiation ,but inhibits enucleation.

| PRJNA797452 | ENA

GATA-1 in proliferating and differentiating murine ES cell derived erythroid progenitors

Project description:This SuperSeries is composed of the following subset Series: GSE35379: Genome-wide occupancy map of GATA-1 in proliferating and differentiating murine ES cell derived erythroid progenitors (ES-EP) GSE35384: Transcriptome analysis of differentiating normal and leukemic erythroid progenitors Refer to individual Series

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Gene expression alterations due to defective Fbw7-dependent cyclin E ubiquitination in bone marrow erythroid cells

Project description:A novel knock-in mouse model was reported (Minella et al., Genes Devel 2008) to have a mixed hyper-proliferative and defective maturation phenotype in the erythroid series. Microarray analyses were performed to identify gene expression alterations resulting from dysregulated cyclin E control. Analysis of gene expression changes in cyclin E (T74A T393A) knock-in erythroblasts (Ter119-positive, CD71-high) isolated by FACS sorting. Comparison is to cells similarly isolated from littermate controls.

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Transcriptome analysis of normal erythroid progenitors and erythroleukemia cells

Project description:We compared the transcriptomes of ES cell derived erythroid progentor cells (ES-EP) and murine erythroleukemia (MEL) cells stably transfected with Gata-1 fused to ER.

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