title: Pre-clinical evaluation of three non-viral gene transfer agents for cystic fibrosis after aerosol delivery to the ovine lung.
abstract: 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.
authors.affiliation: Developmental Biology Division, The Roslin Institute and R(D)SVS, EBVC, University of Edinburgh, Roslin, UK. gerry.mclachlan@ed.ac.uk
authors.forename: G
authors.initials: G
authors.lastname: McLachlan
authors.forename: H
authors.initials: H
authors.lastname: Davidson
authors.forename: E
authors.initials: E
authors.lastname: Holder
authors.forename: L A
authors.initials: LA
authors.lastname: Davies
authors.forename: I A
authors.initials: IA
authors.lastname: Pringle
authors.forename: S G
authors.initials: SG
authors.lastname: Sumner-Jones
authors.forename: A
authors.initials: A
authors.lastname: Baker
authors.forename: P
authors.initials: P
authors.lastname: Tennant
authors.forename: C
authors.initials: C
authors.lastname: Gordon
authors.forename: C
authors.initials: C
authors.lastname: Vrettou
authors.forename: R
authors.initials: R
authors.lastname: Blundell
authors.forename: L
authors.initials: L
authors.lastname: Hyndman
authors.forename: B
authors.initials: B
authors.lastname: Stevenson
authors.forename: A
authors.initials: A
authors.lastname: Wilson
authors.forename: A
authors.initials: A
authors.lastname: Doherty
authors.forename: D J
authors.initials: DJ
authors.lastname: Shaw
authors.forename: R L
authors.initials: RL
authors.lastname: Coles
authors.forename: H
authors.initials: H
authors.lastname: Painter
authors.forename: S H
authors.initials: SH
authors.lastname: Cheng
authors.forename: R K
authors.initials: RK
authors.lastname: Scheule
authors.forename: J C
authors.initials: JC
authors.lastname: Davies
authors.forename: J A
authors.initials: JA
authors.lastname: Innes
authors.forename: S C
authors.initials: SC
authors.lastname: Hyde
authors.forename: U
authors.initials: U
authors.lastname: Griesenbach
authors.forename: E W F W
authors.initials: EW
authors.lastname: Alton
authors.forename: A C
authors.initials: AC
authors.lastname: Boyd
authors.forename: D J
authors.initials: DJ
authors.lastname: Porteous
authors.forename: D R
authors.initials: DR
authors.lastname: Gill
authors.forename: D D S
authors.initials: DD
authors.lastname: Collie
mesh_terms: D000280:Administration, Inhalation; D000818:Animals; D003550:Cystic Fibrosis / Q000235:genetics / Q000628:therapy*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics* / Q000378:metabolism; D018076:DNA, Complementary / Q000008:administration & dosage / Q000235:genetics; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D008081:Liposomes / Q000008:administration & dosage*; D053758:Nanoparticles / Q000008:administration & dosage*; D011092:Polyethylene Glycols; D011094:Polyethyleneimine / Q000008:administration & dosage*; D012333:RNA, Messenger / Q000378:metabolism; D012756:Sheep
chemical_list: C505032:CFTR protein, human; D018076:DNA, Complementary; D008081:Liposomes; D012333:RNA, Messenger; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D011092:Polyethylene Glycols; D011094:Polyethyleneimine