Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Effect of endoglin loss on embryoid bodies

ABSTRACT: Analysis of embryonic sten cell-derived embryoid bodies following endoglin knock out. Loss of endoglin leads to profound reduction of key hematopoietic regulators including SCL, LMO2, Gata2, and TGF-? signaling molecule ALK-1. Results provide insight into molecular mechanisms underlying hemangioblast and primitive hematopoietic development. Total RNA obtained from differentiated day 3 EBs of endoglin knock out ES cells were compared to wild type E14 control ES cells.

ORGANISM(S): Mus musculus

SUBMITTER: Rita Perlingeiro

PROVIDER: E-GEOD-29300 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Modulation of TGF-β signaling by endoglin in murine hemangioblast development and primitive hematopoiesis.

Zhang Liying L Magli Alessandro A Catanese Jacquelyn J Xu Zhaohui Z Kyba Michael M Perlingeiro Rita C R RC

Blood 20110520 1

Endoglin (Eng), an accessory receptor for the transforming growth factor β (TGF-β) superfamily, is required for proper hemangioblast and primitive hematopoietic development. However the mechanism by which endoglin functions at this early developmental stage is currently unknown. Transcriptional analyses of differentiating eng(-/-) and eng(+/+) ES cells revealed that lack of endoglin leads to profound reductions in the levels of key hematopoietic regulators, including Scl, Lmo2, and Gata2. We als ...[more]

PMID: 21602526

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Project description:Molecular mechanisms that regulate the generation of hematopoietic and endothelial cells from mesoderm are poorly understood. To define the underlying mechanisms, we compared gene expression profiles between embryonic stem (ES) cell-derived hemangioblasts (Blast-Colony-Forming Cells, BL-CFCs) and their differentiated progeny, Blast cells. Bioinformatic analysis indicated that BL-CFCs resembled other stem cell populations. A role for Gata2, one of the BL-CFC-enriched transcripts, was further characterized by utilizing the in vitro model of ES cell differentiation. Our studies revealed that Gata2 was a direct target of BMP4 and that enforced GATA2 expression upregulated Bmp4, Flk1 and Scl. Conditional GATA2 induction resulted in a temporal-sensitive increase in hemangioblast generation, precocious commitment to erythroid fate, and increased endothelial cell generation. GATA2 additionally conferred a proliferative signal to primitive erythroid progenitors. Collectively, we provide compelling evidence that GATA2 plays specific, contextual roles in the generation of Flk-1+ mesoderm, the Flk-1+Scl+ hemangioblast, primitive erythroid and endothelial cells. Keywords: Comparison of induced differetiated state to original cell line. Generation of Hemangioblast and Blast Colony cell populations: Scl+/hCD4 ES cells (Chung et al., Development, 2002) were maintained on a layer of mitotically inactivated STO cells in DMEM media in the presence of LIF. Two days before differentiation, the cells were transferred to gelatinized tissue culture flasks and the media was changed to IMDM-based. ES cells were differentiated in liquid-differentiation media in the presence of FCS for 2.75 days. D2.75 EBs were recovered, dissociated in Trypsin/EDTA and stained with antibodies against Flk-1 and human CD4. Cells were then flow sorted on the basis of Flk-1 and Scl expression and the hemangioblast population was considered to be Flk-1+Scl+. D2.75 EB cells were replated as described (Chung et al., 2002) and individual Blast colonies were picked and pooled, and RNA was generated with TRIzol according to the manufacturers protocol. RNA (20 ug) was given to the Washington University Array core were it was reverse transcribed and cRNA synthesized. Replicated of the two populations (hCD4, Blast) were made. Pairwaise comparisons were performed to identify enriched genes.

Dataset Information

Effect of endoglin loss on embryoid bodies

Publications

Modulation of TGF-β signaling by endoglin in murine hemangioblast development and primitive hematopoiesis.

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