Limitations of the murine nose in the development of nonviral airway gene transfer.
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.
Griesenbach
Uta
U
Department of Gene Therapy, Imperial College at the National Heart and Lung Institute, Manresa Road, London SW3 6LR, UK. u.griesenbach@imperial.ac.uk
Sumner-Jones
Stephanie G
SG
Holder
Emma
E
Munkonge
Felix M
FM
Wodehouse
Theresa
T
Smith
Stephen N
SN
Wasowicz
Marguerite Y
MY
Pringle
Ian
I
Casamayor
Isabel
I
Chan
Mario
M
Coles
Rebecca
R
Cornish
Nikki
N
Dewar
Ann
A
Doherty
Ann
A
Farley
Raymond
R
Green
Anne-Marie
AM
Jones
Bryony L
BL
Larsen
Mia D B
MD
Lawton
Anna E
AE
Manvell
Michelle
M
Painter
Hazel
H
Singh
Charanjit
C
Somerton
Lucinda
L
Stevenson
Barbara
B
Varathalingam
Anusha
A
Siegel
Craig
C
Scheule
Ronald K
RK
Cheng
Seng H
SH
Davies
Jane C
JC
Porteous
David J
DJ
Gill
Deborah R
DR
Boyd
A Christopher
AC
Hyde
Steve C
SC
Alton
Eric W F W
EW
D000818:Animals; D001422:Bacterial Adhesion; D003550:Cystic Fibrosis / Q000235:genetics; D003587:Cytomegalovirus / Q000235:genetics; D004742:Enhancer Elements, Genetic; D005260:Female; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D008081:Liposomes / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009154:Mutation; D009666:Nose / Q000473:pathology*; D010957:Plasmids / Q000378:metabolism; D011401:Promoter Regions, Genetic
D008081:Liposomes