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

Gene Ther. 2002 Aug;9(16):1109-15. doi: 10.1038/sj.gt.3301776.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bleomycin
  • Bronchoalveolar Lavage Fluid / immunology
  • Cell Nucleus / metabolism
  • Cytoplasm / metabolism
  • Female
  • Genetic Therapy / methods*
  • Interleukin-6 / biosynthesis
  • Lung / immunology
  • Mice
  • Mice, Inbred C57BL
  • Molecular Mimicry
  • NF-kappa B / genetics*
  • Oligonucleotides / genetics
  • Oligonucleotides / pharmacokinetics*
  • Pneumonia / chemically induced
  • Pneumonia / immunology
  • Pneumonia / prevention & control*
  • Tissue Distribution
  • Transfection

Substances

  • Interleukin-6
  • NF-kappa B
  • Oligonucleotides
  • Bleomycin