Angiogenesis

, Volume 6, Issue 1, pp 31–38

Targeting diphtheria toxin A-chain transcription to activated endothelial cells using an E-selectin promoter

  • Ian H. Maxwell
  • Cortina Kaletta
  • Kurt Naujoks
  • Françoise Maxwell
Article

Abstract

Targeting the transcription of a toxin gene to activated endothelial cells might be used for inhibiting angiogenesis in solid tumors. As a model, we transiently transfected human endothelial cells (HUVEC) in culture with expression plasmids for the toxic A-chain of diphtheria toxin (DT-A), using electroporation (achieving ≈70% transfection efficiency). Protein synthesis in HUVEC was highly sensitive to DT-A expression from constitutive viral promoters. E-selectin is strongly expressed on HUVEC activated by TNFα or TPA. We therefore tested a human E-selectin promoter (−547 to +33) for targeting transcription of DT-A or reporter genes to HUVEC. Luciferase reporters were efficiently expressed in HUVEC from this promoter, with or without an enhancer responsive to Ets-1. Expression was increased by TNFα or TPA. DT-A showed highly preferential expression (increased by TNFα or TPA) in HUVEC, compared with WI38 human fibroblasts. HUVEC expressing DT-A were killed via apoptosis. Overall expression levels were influenced by alternative ‘backbone’ sequences used in the expression plasmids. We propose that delivery of transcriptionally regulated expression plasmids for DT-A in vivo, using cationic lipids that show preferential accumulation in activated or proliferating endothelium, may offer a novel means of inhibiting undesired angiogenesis.

diphtheria toxin endothelial cells E-selectin promoter transcription 

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Copyright information

© Kluwer Academic Publishers 2003

Authors and Affiliations

  • Ian H. Maxwell
    • 1
  • Cortina Kaletta
    • 2
  • Kurt Naujoks
    • 2
  • Françoise Maxwell
    • 3
  1. 1.Department of Dermatology and Cancer CenterUniversity of Colorado Health Sciences CenterDenverUSA
  2. 2.Munich Biotech AGNeuriedGermany
  3. 3.Department of Dermatology and Cancer CenterUniversity of Colorado Health Sciences CenterDenverUSA

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