Coexpression of herpesviral thymidine kinase reporter gene and VEGF gene for noninvasive monitoring of therapeutic gene transfer: an in vitro evaluation. Academic Article uri icon

Overview

MeSH

  • Animals
  • Cell Line
  • Feasibility Studies
  • Gene Expression Profiling
  • Genes, Reporter
  • Rats
  • Recombinant Fusion Proteins

MeSH Major

  • Gene Transfer Techniques
  • Genetic Therapy
  • Muscle Cells
  • Thymidine Kinase
  • Vascular Endothelial Growth Factor A
  • Viral Proteins

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). Accumulation of 14C-2'-fluoro-5-methyl-1-beta-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. 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). 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.

publication date

  • October 2004

has subject area

  • Animals
  • Cell Line
  • Feasibility Studies
  • Gene Expression Profiling
  • Gene Transfer Techniques
  • Genes, Reporter
  • Genetic Therapy
  • Muscle Cells
  • Rats
  • Recombinant Fusion Proteins
  • Thymidine Kinase
  • Vascular Endothelial Growth Factor A
  • Viral Proteins

Research

keywords

  • Journal Article

Identity

Language

  • eng

PubMed ID

  • 15471843

Additional Document Info

start page

  • 1743

end page

  • 1746

volume

  • 45

number

  • 10