Journal article
In vivo imaging of gene transfer to the respiratory tract
Biomaterials, Vol.29(10), pp.1533-1540
2008
DOI: 10.1016/j.biomaterials.2007.11.017
PMID: 18155138
Abstract
Imaging of
in vivo gene expression using luciferase expression in various organs has been used for several years. In contrast to other organs,
in vivo imaging of the lung, particularly after non-viral gene transfer has not been extensively studied. The aim of this study was to address several questions: (1) Does
in vivo light emission correlate with standard tissue homogenate-based luciferase detection in a dose-dependent manner? Recombinant Sendai virus (SeV) transduces airway epithelial cells very efficiently and was used to address this question, (2) Is the sensitivity of the assay sufficient to detect non-viral gene transfer? We treated mice with SeV-Lux vector using our standard “sniffing” protocol, a method that predominantly results in lung deposition. Dose-related
in vivo light emission was visible in all animals. Importantly, there was a significant correlation (
r
>
0.90,
p
<
0.0001) between the
in vivo and
ex vivo assays in both the left and right lung. We next transfected the nasal epithelium via nasal perfusion or the lungs (“sniffing”) of mice with a luciferase plasmid (pCIKLux) complexed to the cationic lipid GL67 (
n
=
25–27/group) and imaged luciferase expression
in vivo 24
h after transfection. Gene expression was detectable in both organs. Correlation between the
in vivo and
ex vivo assays was significant (
r
=
0.52,
p
<
0.005) in the left, but not the right lung. The correlation in the nose was weaker (
r
=
0.45,
p
<
0.05). To our knowledge these studies show for the first time that this non-invasive method of assessing pulmonary gene transfer is viable for evaluating non-viral gene transfer agents.
Details
- Title: Subtitle
- In vivo imaging of gene transfer to the respiratory tract
- Creators
- Uta Griesenbach - Department of Gene Therapy, Imperial College at the National Heart and Lung Institute, Manresa Road, London SW3 6LR, UKCuixiang Meng - Department of Gene Therapy, Imperial College at the National Heart and Lung Institute, Manresa Road, London SW3 6LR, UKRaymond Farley - Department of Gene Therapy, Imperial College at the National Heart and Lung Institute, Manresa Road, London SW3 6LR, UKSeng H Cheng - Genzyme Corporation, Framingham, MA, USARonald K Scheule - Genzyme Corporation, Framingham, MA, USAMark H Davies - Genzyme Ltd, Haverhill, UKPaul C Wolstenholme-Hogg - Genzyme Ltd, Haverhill, UKWillem ten Hove - OctoPlus N.V., Leiden, The NetherlandsPaul van der Hoeven - OctoPlus N.V., Leiden, The NetherlandsPatrick L Sinn - Program in Gene Therapy, Department of Pediatrics, University of Iowa, Iowa City, IA, USAPaul B McCray - Program in Gene Therapy, Department of Pediatrics, University of Iowa, Iowa City, IA, USAMakoto Inoue - DNAVEC Corporation, Tsukuba, JapanDuncan M Geddes - Department of Gene Therapy, Imperial College at the National Heart and Lung Institute, Manresa Road, London SW3 6LR, UKMamoru Hasegawa - DNAVEC Corporation, Tsukuba, JapanGad Frankel - Division of Cell and Molecular Biology, Imperial College London, London, UKSiouxsie Wiles - Division of Cell and Molecular Biology, Imperial College London, London, UKEric W.F.W Alton - Department of Gene Therapy, Imperial College at the National Heart and Lung Institute, Manresa Road, London SW3 6LR, UK
- Resource Type
- Journal article
- Publication Details
- Biomaterials, Vol.29(10), pp.1533-1540
- Publisher
- Elsevier Ltd
- DOI
- 10.1016/j.biomaterials.2007.11.017
- PMID
- 18155138
- ISSN
- 0142-9612
- eISSN
- 1878-5905
- Language
- English
- Date published
- 2008
- Academic Unit
- Microbiology and Immunology; Pulmonary Medicine; Stead Family Department of Pediatrics; Internal Medicine
- Record Identifier
- 9984093507802771
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