Coexpression of herpesviral thymidine kinase reporter gene and VEGF gene for noninvasive monitoring of therapeutic gene transfer: An in vitro evaluation

Martina Anton, Constanze Wittermann, Roland Haubner, Marcus Simoes, Sybille Reder, Bryan Essien, Bettina Wagner, Julia Henke, Wolf Erhardt, Steffi Noll, Neil R. Hackett, Ronald G. Crystal, Markus Schwaiger, Bernd Gansbacher, Frank M. Bengel

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18 Citations (Scopus)

Abstract

Coexpression of a reporter gene and a therapeutic gene may allow for noninvasive monitoring of cardiac gene therapy. We sought to evaluate the usefulness of an adenoviral vector expressing mutant herpesviral thymidine kinase reporter gene (HSV1-sr39tk) and vascular endothelial growth factor (VEGF) 121 in independent expression cassettes (Ad4tk). Methods: Accumulation of 14C-2′-fluoro-5-methyl-1-β-D-arabinofuranosyluracil (FIAU) and 9-(4-18F-fluoro-3-hydroxymethylbutyl)guanine (FHBG) as reporter probes, and secretion of VEGF into medium, were determined for Ad4tk-infected H9c2 rat cardiac cells in vitro. Results: In vitro tracer uptake increased with increasing vector concentration and over time. It was comparable to cells infected with adenovirus expressing only wild-type HSV1-tk (reporter probe: 14C-FIAU) or mutant HSV1-sr39tk (reporter probe: 18F-FHBG) . No significant uptake was observed in cells infected with adenovirus expressing VEGF alone. With increasing vector concentration, Ad4tk-infected cells increasingly released VEGF into medium. VEGF production correlated significantly with cellular reporter probe uptake (r = 0.93; P = 0.0003). Conclusion: The usefulness of a vector coexpressing HSV1-tk and VEGF for noninvasive imaging of expression of a therapeutic transgene has been demonstrated in vitro. This approach may allow for future in vivo monitoring of cardiac angiogenesis gene therapy.

Original languageEnglish
Pages (from-to)1743-1746
Number of pages4
JournalJournal of Nuclear Medicine
Volume45
Issue number10
Publication statusPublished - 1 Oct 2004

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Keywords

  • Gene therapy
  • Heart
  • Hsv1-tk
  • Reporter genes
  • VEGF

ASJC Scopus subject areas

  • Radiology Nuclear Medicine and imaging

Cite this

Anton, M., Wittermann, C., Haubner, R., Simoes, M., Reder, S., Essien, B., Wagner, B., Henke, J., Erhardt, W., Noll, S., Hackett, N. R., Crystal, R. G., Schwaiger, M., Gansbacher, B., & Bengel, F. M. (2004). Coexpression of herpesviral thymidine kinase reporter gene and VEGF gene for noninvasive monitoring of therapeutic gene transfer: An in vitro evaluation. Journal of Nuclear Medicine, 45(10), 1743-1746.