Differential gene expression of human stem progenitor cells derived from early stages of in utero human hematopoiesis

Shojaei, Farbod; Gallacher, Lisa; Bhatia, Mickie

doi:10.1182/blood-2003-09-3209

Cited by 24 publications

(19 citation statements)

References 68 publications

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“…Our results are in line with the observation carried for the primitive CD38 negative subset of CD34+ HSC, which is more abundant in UCB [27]. Even this subpopulation has distinct intrinsic properties depending on the ontological age [28].…”

Section: Fig 3 Transcription Factors Upregulated In Cd133+ Cellssupporting

confidence: 92%

Increased Levels of NOTCH1, NF-κB, and Other Interconnected Transcription Factors Characterize Primitive Sets of Hematopoietic Stem Cells

Panepucci

Oliveira

Zanette

et al. 2010

Stem Cells and Development

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As previously shown, higher levels of NOTCH1 and increased NF-κB signaling is a distinctive feature of the more primitive umbilical cord blood (UCB) CD34+ hematopoietic stem cells (HSCs), as compared to bone marrow (BM). Differences between BM and UCB cell composition also account for this fi nding. The CD133 marker defi nes a more primitive cell subset among CD34+ HSC with a proposed hemangioblast potential. To further evaluate the molecular basis related to the more primitive characteristics of UCB and CD133+ HSC, immunomagnetically purifi ed human CD34+ and CD133+ cells from BM and UCB were used on gene expression microarrays studies. UCB CD34+ cells contained a signifi cantly higher proportion of CD133+ cells than BM (70% and 40%, respectively). Cluster analysis showed that BM CD133+ cells grouped with the UCB cells (CD133+ and CD34+) rather than to BM CD34+ cells. Compared with CD34+ cells, CD133+ had a higher expression of many transcription factors (TFs). Promoter analysis on all these TF genes revealed a signifi cantly higher frequency (than expected by chance) of NF-κB-binding sites (BS), including potentially novel NF-κB targets such as RUNX1, GATA3, and USF1. Selected transcripts of TF related to primitive hematopoiesis and self-renewal, such as RUNX1, GATA3, USF1, TAL1, HOXA9, HOXB4, NOTCH1, RELB, and NFKB2 were evaluated by real-time PCR and were all signifi cantly positively correlated. Taken together, our data indicate the existence of an interconnected transcriptional network characterized by higher levels of NOTCH1, NF-κB, and other important TFs on more primitive HSC sets.

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Section: Fig 3 Transcription Factors Upregulated In Cd133+ Cellssupporting

confidence: 92%

Increased Levels of NOTCH1, NF-κB, and Other Interconnected Transcription Factors Characterize Primitive Sets of Hematopoietic Stem Cells

Panepucci

Oliveira

Zanette

et al. 2010

Stem Cells and Development

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“…To identify potential EKLF target genes, subtractive hybridization was performed with total RNA isolated from day 13.5 fetal livers of wild-type (WT) and EKLF-deficient mice (78). This method has successfully been applied to a large number of applications, including gene expression in cancer, development, and hematopoiesis (28,30,62). In the subtraction utilizing wild-type RNA as the tester population and EKLF-deficient fetal liver RNA as the driver, ϳ175 differentially expressed clones were identified and subjected to sequence analysis.…”

Section: Resultsmentioning

confidence: 99%

Alterations in Expression and Chromatin Configuration of the Alpha Hemoglobin-Stabilizing Protein Gene in Erythroid Krüppel-Like Factor-Deficient Mice

Pilon

Nilson

Zhou

et al. 2006

Molecular and Cellular Biology

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Erythroid Krüppel-like factor (EKLF) is an erythroid zinc finger protein identified by its interaction with a CACCC sequence in the ␤-globin promoter, where it establishes local chromatin structure permitting ␤-globin gene transcription. We sought to identify other EKLF target genes and determine the chromatin status of these genes in the presence and absence of EKLF. We identified alpha hemoglobin-stabilizing protein (AHSP) by subtractive hybridization and demonstrated a 95 to 99.9% reduction in AHSP mRNA and the absence of AHSP in EKLF-deficient cells. Chromatin at the AHSP promoter from EKLF-deficient cells lacked a DNase I hypersensitive site and exhibited histone hypoacetylation across the locus compared to hyperacetylation of wild-type chromatin. Wild-type chromatin demonstrated a peak of EKLF binding over a promoter region CACCC box that differs from the EKLF consensus by a nucleotide. In mobility shift assays, the AHSP promoter CACCC site bound EKLF in a manner comparable to the ␤-globin promoter CACCC site, indicating a broader recognition sequence for the EKLF consensus binding site. The AHSP promoter was transactivated by EKLF in K562 cells, which lack EKLF. These results support the hypothesis that EKLF acts as a transcription factor and a chromatin modulator for the AHSP and ␤-globin genes and indicate that EKLF may play similar roles for other erythroid genes.

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“…The conditions for quantitative real-time (Q-PCR) reactions were reported elsewhere. 16 Forward and reverse primer sequences were 5Ј-gtggctccaggatgttagga-3Ј and 5Ј-gcctgagttcatgttgctga-3Ј for HNF3␣; 5Ј-ccggcagaagattgtagagc-3Ј and 5Ј-cgttggacacgttttgattg-3Ј for Pax-6; and 5Ј-tgcaccaccaactgcttagc-3Ј and 5Ј-ggcatggactgtggtcatgag-3Ј for glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The amount of GOI was normalized to the endogenous housekeeping gene GAPDH.…”

Section: Quantitative Real-time Pcr (Q-pcr)mentioning

confidence: 99%

Human embryonic stem cells maintained in the absence of mouse embryonic fibroblasts or conditioned media are capable of hematopoietic development

Wang

Menéndez

et al. 2005

Blood

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157

109

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Differential gene expression of human stem progenitor cells derived from early stages of in utero human hematopoiesis

Cited by 24 publications

References 68 publications

Increased Levels of NOTCH1, NF-κB, and Other Interconnected Transcription Factors Characterize Primitive Sets of Hematopoietic Stem Cells

Increased Levels of NOTCH1, NF-κB, and Other Interconnected Transcription Factors Characterize Primitive Sets of Hematopoietic Stem Cells

Alterations in Expression and Chromatin Configuration of the Alpha Hemoglobin-Stabilizing Protein Gene in Erythroid Krüppel-Like Factor-Deficient Mice

Human embryonic stem cells maintained in the absence of mouse embryonic fibroblasts or conditioned media are capable of hematopoietic development

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