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Pre-clinical evaluation of three non-viral gene transfer agents for cystic fibrosis after aerosol delivery to the ovine lung


We use both large and small animal models in our pre-clinical evaluation of gene transfer agents (GTAs) for cystic fibrosis (CF) gene therapy. Here, we report the use of a large animal model to assess three non-viral GTAs: 25 kDa-branched polyethyleneimine (PEI), the cationic liposome (GL67A) and compacted DNA nanoparticle formulated with polyethylene glycol-substituted lysine 30-mer. GTAs complexed with plasmids expressing human cystic fibrosis transmembrane conductance regulator (CFTR) complementary DNA were administered to the sheep lung (n=8 per group) by aerosol. All GTAs gave evidence of gene transfer and expression 1 day after treatment. Vector-derived mRNA was expressed in lung tissues, including epithelial cell-enriched bronchial brushing samples, with median group values reaching 1–10% of endogenous CFTR mRNA levels. GL67A gave the highest levels of expression. Human CFTR protein was detected in small airway epithelial cells in some animals treated with GL67A (two out of eight) and PEI (one out of eight). Bronchoalveolar lavage neutrophilia, lung histology and elevated serum haptoglobin levels indicated that gene delivery was associated with mild local and systemic inflammation. Our conclusion was that GL67A was the best non-viral GTA currently available for aerosol delivery to the sheep lung, led to the selection of GL67A as our lead GTA for clinical trials in CF patients.

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This work was funded by the Cystic Fibrosis Trust and CF Trust Senior Fellowships (CB, DRG, UG & GMcL). We thank the animal care and welfare personnel at Easter Bush Veterinary Centre, Roslin, for their assistance.

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Correspondence to G McLachlan.

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RKS is employed by and owns stock and stock options in Genzyme Corporation. SHC is employed by and owns stock and stock options in Genzyme Corporation and has a patent pending on GL-67 assigned to Genzyme. None of the other authors has a financial relationship with a commercial entity that has an interest in the subject of this manuscript.

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McLachlan, G., Davidson, H., Holder, E. et al. Pre-clinical evaluation of three non-viral gene transfer agents for cystic fibrosis after aerosol delivery to the ovine lung. Gene Ther 18, 996–1005 (2011).

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  • gene delivery
  • lung
  • non-viral
  • sheep
  • cystic fibrosis
  • aerosol

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