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

Gene Therapy volume 9, pages 1109–1115 (2002)Cite this article

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 (NFκB) regulates expression of many pro-inflammatory mediators. We have assessed the effect of topical administration of NFκB decoys in a bleomycin model of acute lung inflammation. Using fluorescein-labelled decoy oligonucleotides (ODN) (80 μg/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 μg/mouse did not increase nuclear transfection significantly. We determined the effect of cytoplasmic NFκB decoys on bleomycin-induced inflammation. We transfected mice with ‘naked’ decoy and scrambled ODN (500 μg) 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 NFκB 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.

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Acknowledgements

We thank Dr Naruya Tomita (Osaka University Medical School, Japan) for help in designing the NFκB decoy ODN, Dr Seng Cheng (Genzyme) for providing the lipid GL67 and Mr Ben Booy and Mr Steve Rothery (Confocal Microscopy Unit, Royal Brompton Hospital, London, UK) for help with the confocal analysis. This research was funded by the Cystic Fibrosis Research Trust and a Wellcome Trust Senior Clinical Fellowship (EWFWA).

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  1. Department of Gene Therapy, Faculty of Medicine, Imperial College, London, UK

    U Griesenbach, RL Cassady, RJ Cain, DM Geddes & EWFW Alton

  2. Clinical Genomics Group, Faculty of Medicine, Imperial College, London, UK

    RM duBois

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Griesenbach, U., Cassady, R., Cain, R. et al. Cytoplasmic deposition of NFκB decoy oligonucleotides is insufficient to inhibit bleomycin-induced pulmonary inflammation. Gene Ther 9, 1109–1115 (2002). https://doi.org/10.1038/sj.gt.3301776

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