Limitations of the Murine Nose in the Development of Nonviral Airway Gene Transfer

• DOI: 10.1165/rcmb.2009-0075oc
• Publication Date: 2009-08-01T01:49:05Z
• Authors: Uta Griesenbach; Stephanie G. Sumn...; Emma Holder; Felix M. Munkonge; Theresa Wodehouse; Stephen N. Smith; Marguerite Y. Was...; Ian Pringle; Isabel Casamayor; Mario Chan; Rebecca Coles; Nikki Cornish; Ann Dewar; Ann Doherty; Raymond Farley; Anne-Marie Green; Bryony L. Jones; Mia D. B. Larsen; Anna E. Lawton; Michelle Manvell; Hazel Painter; Charanjit Singh; Lucinda Somerton; Barbara Stevenson; Anusha Varathalingam; Craig Siegel; Ronald K. Scheule; Seng H. Cheng; Jane C. Davies; David J. Porteous; Deborah R. Gill; A. Christopher Boyd; Steve C. Hyde; Eric W. F. W. Alton
• Keywords: Male; Mice, Knockout; 0301 basic medicine; 0303 health sciences; Cystic Fibrosis; Gene Transfer Techniques; Cytomegalovirus; Genetic Therapy; Nose; Bacterial Adhesion; 3. Good health; Mice, Inbred C57BL; Mice; 03 medical and health sciences; Enhancer Elements, Genetic; Liposomes; Mutation; Animals; Female; Promoter Regions, Genetic; Plasmids

A clinical program to assess whether lipid GL67A-mediated gene transfer can ameliorate cystic fibrosis (CF) lung disease is currently being undertaken by the UK CF Gene Therapy Consortium. We have evaluated GL67A gene transfer to the murine nasal epithelium of wild-type and CF knockout mice to assess this tissue as a test site for gene transfer agents. The plasmids used were regulated by either (1) the commonly used short-acting cytomegalovirus promoter/enhancer or (2) the ubiquitin C promoter. In a study of approximately 400 mice with CF, vector-specific CF transmembrane conductance regulator (CFTR) mRNA was detected in nasal epithelial cells of 82% of mice treated with a cytomegalovirus-plasmid (pCF1-CFTR), and 62% of mice treated with an ubiquitin C-plasmid. We then assessed whether CFTR gene transfer corrected a panel of CFTR-specific endpoint assays in the murine nose, including ion transport, periciliary liquid height, and ex vivo bacterial adherence. Importantly, even with the comparatively large number of animals assessed, the CFTR function studies were only powered to detect changes of more than 50% toward wild-type values. Within this limitation, no significant correction of the CF phenotype was detected. At the current levels of gene transfer efficiency achievable with nonviral vectors, the murine nose is of limited value as a stepping stone to human trials.