Coelectrotransfer to skeletal muscle of three plasmids coding for antiangiogenic factors and regulatory factors of the tetracycline-inducible system: tightly regulated expression, inhibition of transplanted tumor growth, and antimetastatic effect

Mol Ther. 2003 Sep;8(3):425-33. doi: 10.1016/s1525-0016(03)00201-6.

Abstract

We describe an approach employing intramuscular plasmid electrotransfer to deliver secretable forms of K1-5 and K1-3-HSA (a fusion of K1-3 with human serum albumin), which span, respectively, five and three of the five kringle domains of plasminogen. A tetracycline-inducible system (Tet-On) composed of three plasmids coding, respectively, for the transgene, the tetracycline transcriptional activator rtTA, and the silencer tTS was employed. K1-3-HSA and K1-5, produced from C2C12 muscle cells, were found to inhibit endothelial cell (HMEC-1) proliferation by 30 and 51%, respectively. In vivo, the expression of the transgene upon doxycycline stimulation was rapid, stable, and tightly regulated (no background expression) and could be maintained for at least 3 months. Blood half-lives of 2.1 and 3.7 days were found for K1-5 and K1-3-HSA, respectively. The K1-5 protein was secreted from muscle into blood at a level of 45 ng/ml, which was sufficient to inhibit MDA-MB-231 tumor growth by 81% in nude mice and B16-F10 melanoma cell lung invasion in C57BL/6 mice by 73%. PECAM-1 immunostaining studies revealed modest tumor vasculature in mice expressing K1-5. In contrast, K1-3-HSA, although secreted into blood at much higher level (250 ng/ml) than K1-5, had no effect on tumor growth.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Angiogenesis Inhibitors / genetics*
  • Angiogenesis Inhibitors / metabolism
  • Electroporation*
  • Gene Expression Regulation
  • Genetic Vectors*
  • Muscle, Skeletal / metabolism*
  • Neoplasm Metastasis / therapy
  • Neoplasms / therapy*
  • Peptides / genetics
  • Peptides / metabolism
  • Plasmids
  • Plasminogen / genetics
  • Plasminogen / metabolism
  • Tetracycline / metabolism
  • Time Factors

Substances

  • Angiogenesis Inhibitors
  • Peptides
  • Plasminogen
  • Tetracycline