Sustained expression of coagulation factor IX by modified cord blood-derived mesenchymal stromal cells

Megan Dodd, Leah Marquez-Curtis, Anna Janowska-Wieczorek, Gonzalo Hortelano

    Research output: Contribution to journalArticle

    5 Citations (Scopus)

    Abstract

    Background: Hemophilia B patients are subject to frequent and spontaneous bleeding caused by a deficiency of clotting factor IX (FIX). Mesenchymal stromal cells (MSCs) have been used in cellular therapies as a result of their immunomodulatory properties, the ability to home to sites of injury and their amenability to various ex vivo modifications, including lentiviral-mediated gene transfer. Methods: MSCs were isolated from human umbilical cord blood and differentiated into adipogenic, chondrogenic and osteogenic lineages. A lentiviral DNA vector containing the human FIX gene was generated using traditional restriction enzyme digest and ligation techniques to generate viable replication-incompetent lentiviral particles that were used to transduce MSCs. Quantitative measurement of FIX expression was conducted using an enzyme-linked immunosorbent assay. Results: The over-expression of FIX was sustained in vitro at levels>4 μg/106 cells/24 h and FIX coagulant activity was>2.5 mIU/106 cells/24 h for the 6-week duration of study. Lentiviral modification of cells with a multiplicity of infection of 10 did not adversely affect the potential of cord blood (CB) MSCs to differentiate to adipocytes, chondrocytes and osteoblastic cells, and the expression of functional FIX was sustained after differentiation and was similar to that in nondifferentiated cells. Conclusions: Modification of human CB MSCs with a lentiviral vector resulted in sustained high FIX expression in vitro after differentiation to adipogenic, chondrogenic and osteoblastic cells. These modified MSCs could have applications in cellular therapies for hemophilia B.

    Original languageEnglish
    Pages (from-to)131-142
    Number of pages12
    JournalJournal of Gene Medicine
    Volume16
    Issue number5-6
    DOIs
    Publication statusPublished - 2014

    Fingerprint

    Factor IX
    Mesenchymal Stromal Cells
    Fetal Blood
    Hemophilia B
    Coagulants
    Blood Coagulation Factors
    Chondrocytes
    Adipocytes
    Genes
    Ligation
    Enzyme-Linked Immunosorbent Assay
    Hemorrhage
    DNA
    Wounds and Injuries
    Enzymes
    Therapeutics
    Infection

    Keywords

    • Coagulation factor IX
    • Hemophilia B
    • Lentiviral vector
    • Mesenchymal stromal cells

    ASJC Scopus subject areas

    • Genetics
    • Molecular Biology
    • Molecular Medicine
    • Genetics(clinical)
    • Drug Discovery
    • Medicine(all)

    Cite this

    Sustained expression of coagulation factor IX by modified cord blood-derived mesenchymal stromal cells. / Dodd, Megan; Marquez-Curtis, Leah; Janowska-Wieczorek, Anna; Hortelano, Gonzalo.

    In: Journal of Gene Medicine, Vol. 16, No. 5-6, 2014, p. 131-142.

    Research output: Contribution to journalArticle

    Dodd, Megan ; Marquez-Curtis, Leah ; Janowska-Wieczorek, Anna ; Hortelano, Gonzalo. / Sustained expression of coagulation factor IX by modified cord blood-derived mesenchymal stromal cells. In: Journal of Gene Medicine. 2014 ; Vol. 16, No. 5-6. pp. 131-142.
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    abstract = "Background: Hemophilia B patients are subject to frequent and spontaneous bleeding caused by a deficiency of clotting factor IX (FIX). Mesenchymal stromal cells (MSCs) have been used in cellular therapies as a result of their immunomodulatory properties, the ability to home to sites of injury and their amenability to various ex vivo modifications, including lentiviral-mediated gene transfer. Methods: MSCs were isolated from human umbilical cord blood and differentiated into adipogenic, chondrogenic and osteogenic lineages. A lentiviral DNA vector containing the human FIX gene was generated using traditional restriction enzyme digest and ligation techniques to generate viable replication-incompetent lentiviral particles that were used to transduce MSCs. Quantitative measurement of FIX expression was conducted using an enzyme-linked immunosorbent assay. Results: The over-expression of FIX was sustained in vitro at levels>4 μg/106 cells/24 h and FIX coagulant activity was>2.5 mIU/106 cells/24 h for the 6-week duration of study. Lentiviral modification of cells with a multiplicity of infection of 10 did not adversely affect the potential of cord blood (CB) MSCs to differentiate to adipocytes, chondrocytes and osteoblastic cells, and the expression of functional FIX was sustained after differentiation and was similar to that in nondifferentiated cells. Conclusions: Modification of human CB MSCs with a lentiviral vector resulted in sustained high FIX expression in vitro after differentiation to adipogenic, chondrogenic and osteoblastic cells. These modified MSCs could have applications in cellular therapies for hemophilia B.",
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