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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.

Authors
  • Martel-Renoir, Dominique1
  • Trochon-Joseph, Véronique
  • Galaup, Ariane
  • Bouquet, Céline
  • Griscelli, Franck
  • Opolon, Paule
  • Opolon, David
  • Connault, Elisabeth
  • Mir, Lluis
  • Perricaudet, Michel
  • 1 Vectorologie et Transfert de Gènes, UMR 8121, Institut Gustave Roussy, 39 Rue Camille Desmoulins, 94805, Villejuif, France. [email protected]
Type
Published Article
Journal
Molecular therapy : the journal of the American Society of Gene Therapy
Publication Date
September 2003
Volume
8
Issue
3
Pages
425–433
Identifiers
PMID: 12946315
Source
Medline
License
Unknown

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.

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