Cytoplasmic deposition of NFkappaB decoy oligonucleotides is insufficient to inhibit bleomycin-induced pulmonary inflammation

U Griesenbach, Robin Cassady-Cain, R J Cain, R M duBois, D M Geddes, E W F W Alton

Research output: Contribution to journalArticlepeer-review

Abstract

Lung inflammation leads to severe tissue destruction and ultimately organ failure in a number of diseases, including cystic fibrosis (CF). The transcription factor nuclear factor kappa B (NFkappaB) regulates expression of many pro-inflammatory mediators. We have assessed the effect of topical administration of NFkappaB decoys in a bleomycin model of acute lung inflammation. Using fluorescein-labelled decoy oligonucleotides (ODN) (80 microg/mouse) we have shown that lipid-complexed and 'naked' ODN transfect conducting airway epithelium in a comparable manner (approximately 65% of cells). However, the ODN were detectable in the cytoplasm, but not in the nucleus of transfected cells. An increase of ODN dose to 500 microg/mouse did not increase nuclear transfection significantly. We determined the effect of cytoplasmic NFkappaB decoys on bleomycin-induced inflammation. We transfected mice with 'naked' decoy and scrambled ODN (500 microg) 1 h before intratracheal administration of bleomycin. We measured IL6 secretion in BALF and lung homogenates and total and differential cell counts in BALF 5 days after bleomycin administration. We did not detect a difference between NFkappaB decoy and scrambled ODN-treated animals in any of the parameters tested. We suggest that access of ODN to the nucleus of airway epithelial cells is a key problem, limiting the efficacy of such decoy strategies, as well as attempts at gene repair.
Original languageEnglish
Pages (from-to)1109-15
Number of pages7
JournalGene Therapy
Volume9
Issue number16
DOIs
Publication statusPublished - Aug 2002

Keywords

  • Animals
  • Bleomycin
  • Bronchoalveolar Lavage Fluid
  • Cell Nucleus
  • Cytoplasm
  • Female
  • Genetic Therapy
  • Interleukin-6
  • Lung
  • Mice
  • Mice, Inbred C57BL
  • Molecular Mimicry
  • NF-kappa B
  • Oligonucleotides
  • Pneumonia
  • Tissue Distribution
  • Transfection

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