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

• Published in: The journal of gene medicine Vol. 16; no. 5-6; pp. 131 - 142
• Main Authors: Dodd, Megan, Marquez-Curtis, Leah, Janowska-Wieczorek, Anna, Hortelano, Gonzalo
• Format: Journal Article
• Language: English
• Published: England Blackwell Publishing Ltd 01.05.2014; Wiley Periodicals Inc
• Subjects: Adipogenesis; Animals; Cell Differentiation; Cell Line; Cells, Cultured; Chondrogenesis; coagulation factor IX; Factor IX - genetics; Factor IX - metabolism; Fetal Blood - cytology; Gene Expression; Gene Order; Gene therapy; Genetic Vectors - genetics; hemophilia B; Hemophilia B - genetics; Hemophilia B - therapy; Humans; lentiviral vector; Lentivirus - genetics; mesenchymal stromal cells; Mesenchymal Stromal Cells - cytology; Mesenchymal Stromal Cells - metabolism; Mice; Partial Thromboplastin Time; Transduction, Genetic; hemophilia B; lentiviral vector; coagulation factor IX; mesenchymal stromal cells
• ISSN: 1099-498X; 1521-2254; 1521-2254
• DOI: 10.1002/jgm.2769

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. Copyright © 2014 John Wiley & Sons, Ltd.