Genetic modification of mesenchymal stem cells to express a single-chain antibody against EGFRvIII on the cell surface

Irina V. Balyasnikova, Rosa Franco-Gou, J. Michael Mathis, Maciej S. Lesniak

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22 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)247-258
Number of pages12
JournalJournal of Tissue Engineering and Regenerative Medicine
Volume4
Issue number4
DOIs
StatePublished - Jan 1 2010

Fingerprint

Single-Chain Antibodies
Stem cells
Mesenchymal Stromal Cells
Adult Stem Cells
Antibodies
Tumors
Gene therapy
Neoplasms
Genetic Therapy
Cells
epidermal growth factor receptor VIII
Antibiotics
Antigens
Drug delivery
Tropism
Medical Genetics
Population Dynamics
Glioblastoma
Microbial Drug Resistance
Brain

Keywords

  • Brain tumours
  • EGFRvIII
  • Mesenchymal stem cells
  • Single-chain antibody
  • Surface
  • Targeting

ASJC Scopus subject areas

  • Medicine (miscellaneous)
  • Biomaterials
  • Biomedical Engineering

Cite this

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title = "Genetic modification of mesenchymal stem cells to express a single-chain antibody against EGFRvIII on the cell surface",
abstract = "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.",
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AU - Balyasnikova, Irina V.

AU - Franco-Gou, Rosa

AU - Michael Mathis, J.

AU - Lesniak, Maciej S.

PY - 2010/1/1

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