Differential gene expression analysis in early and late erythroid progenitor cells in β-thalassaemia

Luke Forster, J. Mccooke, M. Bellgard, David Joske, Jill Finlayson, Reza Ghassemifar

    Research output: Contribution to journalArticle

    6 Citations (Scopus)

    Abstract

    © 2015 John Wiley & Sons Ltd. β- thalassaemia is a disorder of globin gene synthesis resulting in reduced or absent production of the β-globin chain in red blood cells. In this study, haematopoietic stem cells were isolated from the peripheral blood of six transfusion dependent β-thalassaemia patients and six healthy controls. Following 7 and 14d in culture, early- and late- erythroblasts were isolated and purified. No morphological difference in maturation was observed following 7d in culture, while a delayed maturation was observed in the patient group after 14 d. Following RNA isolation and linear amplification, gene expression analyses were performed using microarray technology. The generated data were analysed by two methods: the BRB-ArrayTools platform and the Bioconductor platform using bead level data. Following 7d culture, there was no difference in gene expression between the control and patient groups. Following 14 d culture, 384 differentially expressed genes were identified by either analysis. A subset of 90 genes was selected and the results were confirmed by Quantitative-Real-Time-polymerase chain reaction. Pathways shown to be significantly altered in the patient group include apoptosis, MAPKinase and the nuclear factor-κB pathway.
    Original languageEnglish
    Pages (from-to)257-267
    JournalBritish Journal of Haematology
    Volume170
    Issue number2
    DOIs
    Publication statusPublished - 2015

    Fingerprint

    Erythroid Precursor Cells
    Thalassemia
    Gene Expression
    Globins
    Genes
    Erythroblasts
    Hematopoietic Stem Cells
    Nuclear Family
    Blood Transfusion
    Real-Time Polymerase Chain Reaction
    Erythrocytes
    RNA
    Apoptosis
    Technology
    Control Groups

    Cite this

    Forster, Luke ; Mccooke, J. ; Bellgard, M. ; Joske, David ; Finlayson, Jill ; Ghassemifar, Reza. / Differential gene expression analysis in early and late erythroid progenitor cells in β-thalassaemia. In: British Journal of Haematology. 2015 ; Vol. 170, No. 2. pp. 257-267.
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    abstract = "{\circledC} 2015 John Wiley & Sons Ltd. β- thalassaemia is a disorder of globin gene synthesis resulting in reduced or absent production of the β-globin chain in red blood cells. In this study, haematopoietic stem cells were isolated from the peripheral blood of six transfusion dependent β-thalassaemia patients and six healthy controls. Following 7 and 14d in culture, early- and late- erythroblasts were isolated and purified. No morphological difference in maturation was observed following 7d in culture, while a delayed maturation was observed in the patient group after 14 d. Following RNA isolation and linear amplification, gene expression analyses were performed using microarray technology. The generated data were analysed by two methods: the BRB-ArrayTools platform and the Bioconductor platform using bead level data. Following 7d culture, there was no difference in gene expression between the control and patient groups. Following 14 d culture, 384 differentially expressed genes were identified by either analysis. A subset of 90 genes was selected and the results were confirmed by Quantitative-Real-Time-polymerase chain reaction. Pathways shown to be significantly altered in the patient group include apoptosis, MAPKinase and the nuclear factor-κB pathway.",
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    Differential gene expression analysis in early and late erythroid progenitor cells in β-thalassaemia. / Forster, Luke; Mccooke, J.; Bellgard, M.; Joske, David; Finlayson, Jill; Ghassemifar, Reza.

    In: British Journal of Haematology, Vol. 170, No. 2, 2015, p. 257-267.

    Research output: Contribution to journalArticle

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    T1 - Differential gene expression analysis in early and late erythroid progenitor cells in β-thalassaemia

    AU - Forster, Luke

    AU - Mccooke, J.

    AU - Bellgard, M.

    AU - Joske, David

    AU - Finlayson, Jill

    AU - Ghassemifar, Reza

    PY - 2015

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    N2 - © 2015 John Wiley & Sons Ltd. β- thalassaemia is a disorder of globin gene synthesis resulting in reduced or absent production of the β-globin chain in red blood cells. In this study, haematopoietic stem cells were isolated from the peripheral blood of six transfusion dependent β-thalassaemia patients and six healthy controls. Following 7 and 14d in culture, early- and late- erythroblasts were isolated and purified. No morphological difference in maturation was observed following 7d in culture, while a delayed maturation was observed in the patient group after 14 d. Following RNA isolation and linear amplification, gene expression analyses were performed using microarray technology. The generated data were analysed by two methods: the BRB-ArrayTools platform and the Bioconductor platform using bead level data. Following 7d culture, there was no difference in gene expression between the control and patient groups. Following 14 d culture, 384 differentially expressed genes were identified by either analysis. A subset of 90 genes was selected and the results were confirmed by Quantitative-Real-Time-polymerase chain reaction. Pathways shown to be significantly altered in the patient group include apoptosis, MAPKinase and the nuclear factor-κB pathway.

    AB - © 2015 John Wiley & Sons Ltd. β- thalassaemia is a disorder of globin gene synthesis resulting in reduced or absent production of the β-globin chain in red blood cells. In this study, haematopoietic stem cells were isolated from the peripheral blood of six transfusion dependent β-thalassaemia patients and six healthy controls. Following 7 and 14d in culture, early- and late- erythroblasts were isolated and purified. No morphological difference in maturation was observed following 7d in culture, while a delayed maturation was observed in the patient group after 14 d. Following RNA isolation and linear amplification, gene expression analyses were performed using microarray technology. The generated data were analysed by two methods: the BRB-ArrayTools platform and the Bioconductor platform using bead level data. Following 7d culture, there was no difference in gene expression between the control and patient groups. Following 14 d culture, 384 differentially expressed genes were identified by either analysis. A subset of 90 genes was selected and the results were confirmed by Quantitative-Real-Time-polymerase chain reaction. Pathways shown to be significantly altered in the patient group include apoptosis, MAPKinase and the nuclear factor-κB pathway.

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