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Extensively Self-Renewing Erythroblasts Derived from Transgenic β-Yac Mice are a Novel Model System for Studying Globin Switching and Erythroid Maturation

ABSTRACT: Extensively Self Renewing Erythroblasts (ESREs) are cultured erythroid cells derived from yolk sack or fetal liver. Transcriptome analyses were conducted on ESREs using RNA-seq and compared to published transcriptome analyses of uncultured erythroid cells.

ORGANISM(S): Mus musculus

PROVIDER: GSE54826 | GEO | 2014/10/08

SECONDARY ACCESSION(S): PRJNA237765

REPOSITORIES: GEO

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Extensively Self-Renewing Erythroblasts Derived from Transgenic β-Yac Mice are a Novel Model System for Studying Globin Switching and Erythroid Maturation

Project description:Extensively Self Renewing Erythroblasts (ESREs) are cultured erythroid cells derived from yolk sack or fetal liver. Transcriptome analyses were conducted on ESREs using RNA-seq and compared to published transcriptome analyses of uncultured erythroid cells. RNA-seq was conducted on proliferating Extensively Self Renewing Erythroblasts (ESREs) and ESREs after one day of maturation. Biologic duplicates were done at each time point.

2014-10-08 | E-GEOD-54826 | biostudies-arrayexpress

Extensively self-renewing erythroblasts derived from transgenic ?-yac mice is a novel model system for studying globin switching and erythroid maturation.

Project description:Globin gene regulation occurs in the context of a maturing erythroid cell, which is undergoing significant changes in chromatin structure and gene expression. There are few model systems available that facilitate studies of globin gene regulation in the context of erythroid maturation. Extensively self-renewing erythroblasts (ESREs) are a nontransformed model of erythroid maturation derived from murine fetal liver or yolk sac. Imaging flow cytometry and RNA-seq studies demonstrate that ESREs functionally and molecularly model erythroid maturation. To address the need for a model system that also recapitulates human globin switching, ESREs were derived from mice transgenic for the complete human ?-globin locus (?-yac ESREs). ?-yac ESREs express ?-globin from the transgenic human locus, with minimal ?-globin expression. When treated with hydroxyurea or inhibitors to histone deacetylases, DNA methyltransferases, or the histone demethylase lysine specific demethylase 1 (LSD1), ?-Yac ESREs significantly increase their ?-globin expression, demonstrating their utility for studying agents that influence maturational globin switching. ?-yac ESREs were further used to characterize the secondary effects of LSD1 inhibition on erythroid maturation, with inhibition of LSD1 resulting in altered cell and nuclear size, prolonged Kit expression, and decreased rates of enucleation consistent with impaired maturation. Taken together, these studies demonstrate that ?-yac ESREs have significant utility for identifying modulators of maturational globin switching as well as for studying the broader role of those modulators in erythroid maturation.

| S-EPMC6176860 | biostudies-other

Cannibalized Erythroblasts Accelerate Developmental Neurogenesis by Regulating Mitochondrial Dynamics

Project description:In this study, we disclose a mechanism triggered during neurulation that programs brain development by donation of sacrificial yolk sac erythroblasts to neuroepithelial cells. At embryonic day (E) 8.5, neuroepithelial cells transiently integrate with the endothelium of yolk sac blood vessels and cannibalize intravascular erythroblasts as transient heme-rich endosymbionts. This cannibalistic behavior instructs precocious neuronal differentiation of neuroepithelial cells in the proximity of blood vessels. By experiments in vitro, we show that access to erythroblastic heme accelerates the pace of neurogenesis by induction of a truncated neurogenic differentiation program from a poised state.

2021-03-03 | GSE167515 | GEO

Erythroblasts

Project description:Erythroblasts

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Transcriptional profiling of primary human erythroblasts infected with Plasmodium falciparum

Project description:In malaria infection, Plasmodium spp. parasites accumulate in the bone marrow near sites of erythroid development. While it has been observed that Plasmodium falciparum infection of late-stage erythroblasts can delay terminal erythroid differentiation and enucleation, the mechanism(s) underlying this phenomenon are unknown. Here, we apply RNA-seq after fluorescence-activated cell sorting (FACS) of infected erythroblasts to identify transcriptional responses to direct and indirect interaction with P. falciparum.

2023-07-25 | GSE233739 | GEO

RNA-seq analysis of differentiating human erythroblasts

Project description:Human erythroblasts purified from cord blood were cultured in vitro and FACS-sorted into five highly purified populations representing distinct differentiation stages: proerythroblasts, early basophilic erythroblasts, late basophilic erythroblasts, polychromatophilic erythroblasts, and orthochromatophilic erythroblasts. The methods for culture and sorting experiments are given in Hu et al. 2013. For each RNA-seq library, RNA was isolated from 1x 106 sorted human erythroblasts using RNeasy Plus Mini kits (Qiagen). Libraries were then prepared using Illumina TruSeqTM RNA kits to obtain 50 nt reads. Collaborators at the New Your Blood Center were responsible for erythroblast culture, FACS purification of erythroblast populations, and acquisition of RNA-seq data. Collaborators at U.C. Berkeley and Lawrence Berkeley National Laboratory performed data analysis and experimental validation of alternative splicing in erythroblasts. Results: Differentiating erythroblasts execute a dynamic alternative splicing program that is enriched in genes affecting cell cycle, organelle organization, chromatin function, and RNA processing. Alternative splicing plays a major role in regulating gene expression to ensure synthesis of appropriate proteome at each stage as the cells remodel in preparation for production of mature red cells. Erythroid differentiation stage-specific transcriptome analysis was performed by RNA-seq analysis of highly purified erythroblast populations

2013-12-25 | E-GEOD-53635 | biostudies-arrayexpress

RNA-Seq analysis of microRNA expression profiles in mouse primary CFU-E late erythroid progenitors and Ter119+ mature erythroblasts

Project description:Using RNA-seq technology, we quantitatively determined the expression profile of microRNAs during mouse terminal erythroid differentiation. CFU-E erythroid progenitors were isolated from E14.5 fetal liver as the Ter119, B220, Mac-1, CD3 and Gr-1 negative, C-Kit positive and 20% high CD71 population. Mature Ter119+ erythroblasts were isolated from E14.5 fetal liver as C-Kit negative and Ter119 positive population. Consistent with nuclear condensation and global gene expression shut down during terminal erythroid differentiation, we found that the majority of microRNAs are downregulated in more mature Ter119+ erythroblasts compared with CFU-E erythroid progenitors. Examination of microRNA expression profiles in 2 cell types

2011-06-09 | E-GEOD-25602 | biostudies-arrayexpress

Ribosomal deficiencies in Diamond-Blackfan anemia impair translation of transcripts essential for differentiation of murine and human erythroblasts

Project description:Diamond Blackfan Anemia (DBA) is associated with developmental defects and profound anemia. Mutations in genes encoding a ribosomal protein of the small (e.g. Rps19) or large (e.g. Rpl11) ribosomal subunit are found in over half of these patients. The mutations cause ribosomal haploinsufficiency, which reduces overall translation efficiency of cellular mRNAs. We reduced expression of *Rps19* or *Rpl11* in mouse erythroblasts and investigated mRNA polyribosome association, which revealed deregulated translation initiation of specific transcripts. Among these were *Bag1*, encoding a Hsp70 co-chaperone, and *Csde1*, encoding an RNA binding protein, both expressed at increased levels in erythroblasts. Their translation initiation is cap-independent and starts from an internal ribosomal entry site (IRES), which appeared sensitive to knock down of Rps19 or Rpl11. Mouse embryos lacking Bag1 die at embryonic day E13.5 with reduced erythroid colony forming cells in the fetal liver, and low Bag1 expression impairs erythroid differentiation in vitro. Reduced expression of Csde1 impairs proliferation and differentiation of erythroid blasts. Protein but not mRNA expression of *BAG1* and *CSDE1* was reduced in erythroblasts cultured from DBA patients. Our data suggest that impaired IRES-mediated translation of mRNAs expressed at increased levels in erythroblasts contributes to the erythroid phenotype of DBA. 3 biological replicates of erythroblasts treated with different shRNA were used for polyribosomal sucrose gradients; RNA was extracted from gradients in 2 samples - mRNA associated with polyribosomes (poly) and the rest (sub).

2011-11-14 | E-GEOD-22903 | biostudies-arrayexpress

RNA-Seq analysis of microRNA expression profiles in mouse primary CFU-E late erythroid progenitors and Ter119+ mature erythroblasts

2011-06-09 | GSE25602 | GEO

14q32 and let-7 microRNAs regulate transcriptional networks in fetal and adult human erythroblasts

Project description:In humans, fetal erythropoiesis takes place in the liver whereas adult erythropoiesis occurs in the bone marrow. Fetal and adult erythroid cells are not only produced at different sites, but are also distinguished by their respective transcriptional program. In particular, whereas fetal erythroid cells express γ-globin chains to produce fetal hemoglobin (HbF), adult cells express β-globin chains to generate adult hemoglobin. Understanding the transcriptional regulation of the fetal-to-adult hemoglobin switch is clinically important as re-activation of HbF production in adult erythroid cells would represent a promising therapy for the hemoglobin disorders sickle cell disease and β-thalassemia. We used RNA-sequencing to measure global gene and microRNA (miRNA) expression in human erythroblasts derived ex vivo from fetal liver (N = 12 donors) and bone marrow (N = 12 donors) hematopoietic stem/progenitor cells. We identified 7,829 transcripts and 402 miRNA that were differentially expressed (false discovery rate (FDR) <5%). The miRNA expression patterns were replicated in an independent collection of human erythroblasts using a different technology. By combining gene and miRNA expression data, we developed transcriptional networks which show substantial differences between fetal and adult human erythroblasts. Our analyses highlighted the miRNAs at the imprinted 14q32 locus in fetal erythroblasts and the let-7 miRNA family in adult erythroblasts as key regulators of stage-specific erythroid transcriptional programs. Altogether, our results provide a comprehensive resource to prioritize genes that may modify clinical severity in red blood cell (RBC) disorders, or genes that might be implicated in erythropoiesis by genome-wide association studies of RBC traits.

2018-02-22 | GSE110936 | GEO

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