Fetal expression of a human Aγ globin transgene rescues globin chain imbalance but not hemolysis in EKLF null mouse embryos

Perkins, Andrew; Peterson, K. R.; Stamatoyannopoulos, George; Witkowska, H. Ewa; Orkin, Stuart H.

doi:10.1182/blood.v95.5.1827.004k10_1827_1833

Cited by 53 publications

(20 citation statements)

References 29 publications

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“…In studies involving knockout mice, the erythroid specific CACCC-binding transcription factor erythroid kruppel-like factor (EKLF) appeared to be necessary for beta globin expression (80,81). However, beta globin is not the only target of EKLF, since transgene expression of human Ag globin and efficient production of mouse/human hybrid hemoglobin did not rescue hemolysis in EKLF -/mouse embryos (82). The cDNA array screen revealed a rapid accumulation of the already highly abundant accumulation EKLF transcripts upon differentiation induction (6 and 20 h), followed by a decrease at 48 h (Fig.…”

Section: Signal Transducers and Transcription Factors Differentially mentioning

confidence: 99%

Establishment of normal, terminally differentiating mouse erythroid progenitors: molecular characterization by cDNA arrays

Dolznig

Boulmé²,

Stangl³

et al. 2001

FASEB j.

121

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Expression profiling with cDNA arrays is an excellent tool for molecular analysis of complex processes such as terminal erythroid differentiation. The shortcomings of the currently available erythroid in vitro differentiation models, however, severely impaired the usefulness of this approach to study erythropoiesis. Here, we describe a novel, murine erythroid cell system closely corresponding to in vivo erythroid progenitors. Mortal, long‐term proliferating erythroid progenitors of fetal liver or immortal strains of p53‐deficient erythroblasts were established in culture. Both cell types proliferated in serum‐free medium and were strictly dependent on physiologically relevant cytokines and hormones, stably retaining a diploid set of chromosomes. If exposed to physiological differentiation factors (erythropoietin plus insulin), cells synchronously recapitulated the normal in vivo differentiation program to mature terminally into enucleated erythrocytes and expressed stage‐specific erythroid transcription factors in the expected temporal order. Using cDNA arrays, we found a large number of genes differentially expressed at time points during differentiation. Already 6 h after differentiation induction, 17% of the expressed genes showed significant alterations in mRNA abundance, increasing to 53% (12% up‐regulated, 41% down‐regulated genes) by 48 h. Cluster analysis of mRNA expression kinetics during differentiation identified six distinct expression patterns. All genes on the array with a known function in erythropoiesis showed the expected variations in expression. The genes identified also allowed first insights into the sequence of events within the regulatory network responsible for erythroid maturation. In mortal wild‐type as well as immortal p53‐/‐ erythroblasts, changes in mRNA abundance of several well‐regulated gene products was verified at the protein level. Taken together, this novel hematopoietic cell system faithfully executes essential steps of normal erythropoiesis and allows us to dissect and characterize molecular mechanisms involved in erythropoiesis.

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Section: Signal Transducers and Transcription Factors Differentially mentioning

confidence: 99%

Establishment of normal, terminally differentiating mouse erythroid progenitors: molecular characterization by cDNA arrays

Dolznig

Boulmé²,

Stangl³

et al. 2001

FASEB j.

121

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“…The Krüppel gene is responsible for segmentation in the developing fruit fly, and many of its mammalian counterparts are known to play roles in regulating cell proliferation, differentiation, and development (1). For instance, one of the first identified KLF, erythroid KLF (KLF1), has been shown to be a critical activator of the ␤-globin gene in erythroid cells and appears to be required for normal adult erythropoiesis (2,(30)(31)(32)(33). Likewise, the early growth response-␣/transforming growth factor-␤ inducible early gene 1 (KLF10) has been implicated in mediating cell cycle arrest and apoptosis (3,15,39,41,42).…”

mentioning

confidence: 99%

Enterocyte differentiation marker intestinal alkaline phosphatase is a target gene of the gut-enriched Krüppel-like factor

Hinnebusch

Siddique

Henderson

et al. 2004

American Journal of Physiology-Gastrointestinal and Liver Physi

114

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We have examined the role that the transcription factor gut-enriched Krüppel-like factor (KLF4 or GKLF) plays in activating the enterocyte differentiation marker gene intestinal alkaline phosphatase (IAP). A yeast one-hybrid screen was used to identify proteins interacting with a previously identified cis-element (IF-III) located within the human IAP gene promoter. DNA-protein interactions were determined by using EMSA. Northern blot analysis was used to study RNA expression in human colon cancer RKO cells engineered to overexpress KLF4. Transient transfections with IAP-luciferase reporter constructs were used to characterize the mechanisms by which KLF4 activates IAP transcription. The yeast one-hybrid screen and EMSA identified KLF4 as binding to IF-III. RKO cells induced to overexpress KLF4 demonstrated a corresponding dose-dependent increase in IAP expression, and EMSA with nuclear extract from these cells confirmed that KLF4 binds to the IF-III element. Transient transfections revealed that KLF4 transactivated the IAP gene largely via a critical segment in the IAP promoter that includes the IF-III cis-element. Mutant KLF4 constructs failed to fully activate IAP. We have identified the enterocyte differentiation marker IAP as a KLF4 target gene. IAP transactivation by KLF4 is likely mediated through a critical region located within the proximal IAP promoter region.

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“…Klf1 knockout mice are embryonic lethal and die of anaemia (Perkins et al, 1995). Although transgenic b-YAC carrying Klf1 knockout mice have elevated HBG expression in the fetal liver, this is not sufficient to compensate for absent adult globin (Perkins et al, 2000). Numerous human KLF1 variants have been described with a variety of erythroid phenotypes (Perkins et al, 2016).…”

Section: Review ª 2017 John Wiley and Sons Ltdmentioning

confidence: 99%

Recent progress in understanding and manipulating haemoglobin switching for the haemoglobinopathies

2017

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The major β-haemoglobinopathies, sickle cell disease and β-thalassaemia, represent the most common monogenic disorders worldwide and a steadily increasing global disease burden. Allogeneic haematopoietic stem cell transplantation, the only curative therapy, is only applied to a small minority of patients. Common clinical management strategies act mainly downstream of the root causes of disease. The observation that elevated fetal haemoglobin expression ameliorates these disorders has motivated longstanding investigations into the mechanisms of haemoglobin switching. Landmark studies over the last decade have led to the identification of two potent transcriptional repressors of γ-globin, BCL11A and ZBTB7A. These regulators act with additional trans-acting epigenetic repressive complexes, lineage-defining factors and developmental programs to silence fetal haemoglobin by working on cis-acting sequences at the globin gene loci. Rapidly advancing genetic technology is enabling researchers to probe deeply the interplay between the molecular players required for γ-globin (HBG1/HBG2) silencing. Gene therapies may enable permanent cures with autologous modified haematopoietic stem cells that generate persistent fetal haemoglobin expression. Ultimately rational small molecule pharmacotherapies to reactivate HbF could extend benefits widely to patients.

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Fetal expression of a human Aγ globin transgene rescues globin chain imbalance but not hemolysis in EKLF null mouse embryos

Cited by 53 publications

References 29 publications

Establishment of normal, terminally differentiating mouse erythroid progenitors: molecular characterization by cDNA arrays

Establishment of normal, terminally differentiating mouse erythroid progenitors: molecular characterization by cDNA arrays

Enterocyte differentiation marker intestinal alkaline phosphatase is a target gene of the gut-enriched Krüppel-like factor

Recent progress in understanding and manipulating haemoglobin switching for the haemoglobinopathies

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