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Genetic modification of mesenchymal stem cells to express a single‐chain antibody against EGFRvIII on the cell surface
Author(s) -
Balyasnikova Irina V.,
FrancoGou Rosa,
Mathis J. Michael,
Lesniak Maciej S.
Publication year - 2010
Publication title -
journal of tissue engineering and regenerative medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.835
H-Index - 72
eISSN - 1932-7005
pISSN - 1932-6254
DOI - 10.1002/term.228
Subject(s) - mesenchymal stem cell , transfection , antibody , nucleofection , cell , chemistry , population , cell culture , tropism , microbiology and biotechnology , cancer research , biology , immunology , medicine , genetics , biochemistry , virus , environmental health
Human adult mesenchymal stem cells (hMSCs) are under active investigation as cellular carriers for gene therapy. hMSCs possess natural tropism toward tumours; however, the targeting of hMSCs to specific cell populations within tumours is unexplored. In the case of glioblastoma multiforme (GBM), at least half of the tumours express EGFRvIII on the cell surface, an ideal target for antibody‐mediated gene/drug delivery. In this study, we investigated the feasibility of genetically modifying hMSCs to express a single‐chain antibody (scFv) to EGFRvIII on their surfaces. Nucleofection was used to transfect hMSCs with cDNA encoding scFv EGFRvIII fused with PDGFR or human B7‐1 transmembrane domains. The expression of scFv EGFRvIII on the cell surface was assessed by FACS. A stable population of scFv EGFRvIII‐expressing hMSCs was selected, based on antibiotic resistance, and enriched using FACS. We found that nucleofection allows the efficient expression of scFv EGFRvIII on the cell surface of hMSCs. hMSCs transfected with the construct encoding scFv EGFRvIII as a fusion with PDGFRtm showed scFv EGFRvIII expression in up to 86% of cells. Most importantly, human MSCs expressing scFv against EGFRvIII demonstrated enhanced binding to U87‐EGFRvIII cells in vitro and significantly increased retention in human U87‐EGFRvIII‐expressing tumours in vivo . In summary, we provide the first conclusive evidence of genetic modification of hMSCs with a single‐chain antibody against an antigen expressed on the surface of tumour cells, thereby opening up a new venue for enhanced delivery of gene therapy applications in the context of malignant brain cancer. Copyright © 2009 John Wiley & Sons, Ltd.