• Login
    View Item 
    •   Home
    • Colleges & Programs
    • Medical College of Georgia (MCG)
    • Department of Pediatrics
    • Department of Pediatrics: Faculty Research and Presentations
    • View Item
    •   Home
    • Colleges & Programs
    • Medical College of Georgia (MCG)
    • Department of Pediatrics
    • Department of Pediatrics: Faculty Research and Presentations
    • View Item
    JavaScript is disabled for your browser. Some features of this site may not work without it.

    Browse

    All of Scholarly CommonsCommunitiesTitleAuthorsIssue DateSubmit DateSubjectsThis CollectionTitleAuthorsIssue DateSubmit DateSubjects

    My Account

    LoginRegister

    About

    AboutCreative CommonsAugusta University LibrariesUSG Copyright Policy

    Statistics

    Display statistics

    Characterization of the transcriptome profiles related to globin gene switching during in vitro erythroid maturation

    • CSV
    • RefMan
    • EndNote
    • BibTex
    • RefWorks
    Thumbnail
    Name:
    1471-2164-13-153.pdf
    Size:
    1.086Mb
    Format:
    PDF
    Download
    Authors
    Li, Biaoru
    Ding, Lianghao
    Li, Wei
    Story, Michael D
    Pace, Betty S.
    Issue Date
    2012-04-26
    URI
    http://hdl.handle.net/10675.2/790
    
    Metadata
    Show full item record
    Abstract
    Background: The fetal and adult globin genes in the human b-globin cluster on chromosome 11 are sequentially expressed to achieve normal hemoglobin switching during human development. The pharmacological induction of fetal g-globin (HBG) to replace abnormal adult sickle bS-globin is a successful strategy to treat sickle cell disease; however the molecular mechanism of g-gene silencing after birth is not fully understood. Therefore, we performed global gene expression profiling using primary erythroid progenitors grown from human peripheral blood mononuclear cells to characterize gene expression patterns during the g-globin to b-globin (g/b) switch observed throughout in vitro erythroid differentiation
    Results: We confirmed erythroid maturation in our culture system using cell morphologic features defined by Giemsa staining and the g/b-globin switch by reverse transcription-quantitative PCR (RT-qPCR) analysis. We observed maximal gglobin expression at day 7 with a switch to a predominance of b-globin expression by day 28 and the g/b-globin switch occurred around day 21. Expression patterns for transcription factors including GATA1, GATA2, KLF1 and NFE2 confirmed our system produced the expected pattern of expression based on the known function of these factors in globin gene regulation. Subsequent gene expression profiling was performed with RNA isolated from progenitors harvested at day 7, 14, 21, and 28 in culture. Three major gene profiles were generated by Principal Component Analysis (PCA). For profile-1 genes, where expression decreased from day 7 to day 28, we identified 2,102 genes downregulated > 1.5-fold. Ingenuity pathway analysis (IPA) for profile-1 genes demonstrated involvement of the Cdc42, phospholipase C, NF-Kb, Interleukin-4, and p38 mitogen activated protein kinase (MAPK) signaling pathways. Transcription factors known to be involved in g-and b-globin regulation were identified. The same approach was used to generate profile-2 genes where expression was up-regulated over 28 days in culture. IPA for the 2,437 genes with > 1.5-fold induction identified the mitotic roles of polo-like kinase, aryl hydrocarbon receptor, cell cycle control, and ATM (Ataxia Telangiectasia Mutated Protein) signaling pathways; transcription factors identified included KLF1, GATA1 and NFE2 among others. Finally, profile-3 was generated from 1,579 genes with maximal expression at day 21, around the time of the g/b-globin switch. IPA identified associations with cell cycle control, ATM, and aryl hydrocarbon receptor signaling pathways.
    Conclusions: The transcriptome analysis completed with erythroid progenitors grown in vitro identified groups of genes with distinct expression profiles, which function in metabolic pathways associated with cell survival, hematopoiesis, blood cells activation, and inflammatory responses. This study represents the first report of a transcriptome analysis in human primary erythroid progenitors to identify transcription factors involved in hemoglobin switching. Our results also demonstrate that the in vitro liquid culture system is an excellent model to define mechanisms of global gene expression and the DNA-binding protein and signaling pathways involved in globin gene regulation.
    Citation
    BMC Genomics. 2012 Apr 26; 13:153
    ae974a485f413a2113503eed53cd6c53
    10.1186/1471-2164-13-153
    Scopus Count
    Collections
    Department of Pediatrics: Faculty Research and Presentations

    entitlement

    Related articles

    • Characterization of transcription factor networks involved in umbilical cord blood CD34+ stem cells-derived erythropoiesis.
    • Authors: Li B, Ding L, Yang C, Kang B, Liu L, Story MD, Pace BS
    • Issue date: 2014
    • Plastrum testudinis induces γ-globin gene expression through epigenetic histone modifications within the γ-globin gene promoter via activation of the p38 MAPK signaling pathway.
    • Authors: Qian X, Chen J, Zhao D, Guo L, Qian X
    • Issue date: 2013 Jun
    • Mi2β-mediated silencing of the fetal γ-globin gene in adult erythroid cells.
    • Authors: Amaya M, Desai M, Gnanapragasam MN, Wang SZ, Zu Zhu S, Williams DC Jr, Ginder GD
    • Issue date: 2013 Apr 25
    • Globin gene expression in correlation with G protein-related genes during erythroid differentiation.
    • Authors: Čokić VP, Smith RD, Biancotto A, Noguchi CT, Puri RK, Schechter AN
    • Issue date: 2013 Feb 20
    • Multiple physical stresses induce γ-globin gene expression and fetal hemoglobin production in erythroid cells.
    • Authors: Schaeffer EK, West RJ, Conine SJ, Lowrey CH
    • Issue date: 2014 Apr
    DSpace software (copyright © 2002 - 2023)  DuraSpace
    Quick Guide | Contact Us
    Open Repository is a service operated by 
    Atmire NV
     

    Export search results

    The export option will allow you to export the current search results of the entered query to a file. Different formats are available for download. To export the items, click on the button corresponding with the preferred download format.

    By default, clicking on the export buttons will result in a download of the allowed maximum amount of items.

    To select a subset of the search results, click "Selective Export" button and make a selection of the items you want to export. The amount of items that can be exported at once is similarly restricted as the full export.

    After making a selection, click one of the export format buttons. The amount of items that will be exported is indicated in the bubble next to export format.