Please use this identifier to cite or link to this item: http://hdl.handle.net/2440/96079
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Type: Journal article
Title: Transduction of ferret airway epithelia using a pre-treatment and lentiviral gene vector
Author: Cmielewski, P.
Farrow, N.
Donnelley, M.
McIntyre, C.
Penny-Dimri, J.
Kuchel, T.
Parsons, D.
Citation: BMC Pulmonary Medicine, 2014; 14(1):183-1-183-7
Publisher: BioMed Central
Issue Date: 2014
ISSN: 1471-2466
1471-2466
Statement of
Responsibility: 
Patricia Cmielewski, Nigel Farrow, Martin Donnelley, Chantelle McIntyre, Jahan Penny-Dimri, Tim Kuchel and David Parsons
Abstract: BACKGROUND: The safety and efficiency of gene therapies for cystic fibrosis (CF) need to be assessed in pre-clinical models. Using the normal ferret, this study sought to determine whether ferret airway epithelia could be transduced with a lysophosphatidylcholine (LPC) pre-treatment followed by a VSV-G pseudotyped HIV-1 based lentiviral (LV) vector, in preparation for future studies in CF ferrets. METHODS: Six normal ferrets (7 -8 weeks old) were treated with a 150 μL LPC pre-treatment, followed one hour later by a 500 μL LV vector dose containing the LacZ transgene. LacZ gene expression in the conducting airways and lung was assessed by X-gal staining after 7 days. The presence of transduction in the lung, as well as off-target transduction in the liver, spleen and gonads, were assessed by qPCR. The levels of LV vector p24 protein bio-distribution in blood sera were assessed by ELISA at 0, 1, 3, 5 and 7 days. RESULTS: The dosing protocol was well tolerated. LacZ gene expression was observed en face in the trachea of all animals. Histology showed that ciliated and basal cells were transduced in the trachea, with rare LacZ transduced single cells noted in lung. p24 levels was not detectable in the sera of 5 of the 6 animals. The LacZ gene was not detected in the lung tissue and no off-target transduction was detected by qPCR. CONCLUSIONS: This study shows that ferret airway epithelia are transducible using our unique two-step pre-treatment and LV vector dosing protocol. We have identified a number of unusual anatomical factors that are likely to influence the level of transduction that can be achieved in ferret airways. The ability to transduce ferret airway epithelium is a promising step towards therapeutic LV-CFTR testing in a CF ferret model.
Keywords: Ferret; Lung; Lentivirus; Cystic fibrosis; Gene therapy
Rights: © 2014 Cmielewski et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
RMID: 0030018364
DOI: 10.1186/1471-2466-14-183
Appears in Collections:Medicine publications

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