Concurrent delivery of GM-CSF and endostatin genes by a single adenoviral vector provides a synergistic effect on the treatment of orthotopic liver tumors

J Gene Med. 2003 May;5(5):386-98. doi: 10.1002/jgm.376.

Abstract

Background: The immune resistance of large tumors represents a major problem for cancer immunotherapy, whereas the need for repeated injections of high doses of recombinant anti-angiogenic proteins represents a similar problem for anti-angiogenic therapy. To test whether antitumor activity could be increased by combining the above two mechanisms, this study examined the therapeutic effect of combination gene therapy using a murine granulocyte-macrophage colony-stimulating factor (mGM-CSF) gene and a human endostatin (hED) gene on a rat orthotopic liver tumor model.

Methods: An adenoviral vector was constructed that simultaneously carried two transcriptional cassettes, for the expression of mGM-CSF and hED, respectively, or that carried a single cassette of either gene. The adenoviruses were intratumorally administered to 3-day-old or 7-day-old tumors. Moreover, the antitumor effects of the combination therapy and monotherapy were assessed and compared.

Results: The double-gene-containing adenoviral vector expressed transgenes as efficiently as the single-gene-containing vector. Moreover, the adenovirally expressed endostatin was biologically active, as demonstrated in vitro and in vivo. Results from animal experiments demonstrated a synergistic antitumor effect induced by the combined mGM-CSF and hED therapy. The combination of hED with mGM-CSF enhanced tumor-specific CTL activity, but did not interfere with the infiltration of cellular effectors in the tumor regions. The blood vessel density of the liver tumors markedly reduced as a result of hED expression in both monotherapy and combination therapy. Furthermore, combination therapy significantly increased the number of apoptotic cells in the tumor regions.

Conclusions: The experimental results suggest that the combined gene therapy against tumor cells and the tumor vascular system using antitumor immune mechanisms and anti-angiogenic mechanisms holds promise as a strategy for treating cancers.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviridae / genetics*
  • Angiogenesis Inhibitors / pharmacology
  • Animals
  • Apoptosis
  • Blotting, Western
  • Cell Division
  • Cell Line
  • Cell Line, Tumor
  • Collagen / pharmacology
  • Combined Modality Therapy
  • Drug Combinations
  • Endostatins / genetics*
  • Endothelial Cells / cytology
  • Enzyme-Linked Immunosorbent Assay
  • Gene Transfer Techniques
  • Genetic Therapy / methods*
  • Genetic Vectors
  • Granulocyte-Macrophage Colony-Stimulating Factor / genetics*
  • Granulocyte-Macrophage Colony-Stimulating Factor / metabolism
  • Humans
  • Immunohistochemistry
  • Immunotherapy / methods*
  • In Situ Nick-End Labeling
  • Laminin / pharmacology
  • Liver / metabolism
  • Liver Neoplasms / genetics*
  • Liver Neoplasms / metabolism
  • Mice
  • Mice, Inbred BALB C
  • Models, Genetic
  • Neovascularization, Pathologic
  • Plasmids / metabolism
  • Platelet Endothelial Cell Adhesion Molecule-1 / biosynthesis
  • Proteoglycans / pharmacology
  • Rats
  • Rats, Inbred F344
  • Reverse Transcriptase Polymerase Chain Reaction
  • T-Lymphocytes, Cytotoxic / metabolism
  • Time Factors
  • Transcription, Genetic
  • Transgenes

Substances

  • Angiogenesis Inhibitors
  • Drug Combinations
  • Endostatins
  • Laminin
  • Platelet Endothelial Cell Adhesion Molecule-1
  • Proteoglycans
  • matrigel
  • Granulocyte-Macrophage Colony-Stimulating Factor
  • Collagen