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Restoring ciliary function to differentiated primary ciliary dyskinesia cells with a lentiviral vector

Gene Therapy volume 21, pages 253–261 (2014)Cite this article

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Abstract

Primary ciliary dyskinesia (PCD) is a genetically heterogenous autosomal recessive disease in which mutations disrupt ciliary function, leading to impaired mucociliary clearance and life-long lung disease. Mouse tracheal cells with a targeted deletion in the axonemal dynein intermediate chain 1 (Dnaic1) gene differentiate normally in culture but lack ciliary activity. Gene transfer to undifferentiated cultures of mouse Dnaic1−/− cells with a lentiviral vector pseudotyped with avian influenza hemagglutinin restored Dnaic1 expression and ciliary activity. Importantly, apical treatment of well-differentiated cultures of mouse Dnaic1−/− cells with lentiviral vector also restored ciliary activity, demonstrating successful gene transfer from the apical surface. Treatment of Dnaic1flox/flox mice expressing an estrogen-responsive Cre recombinase with different doses of tamoxifen indicated that restoration of ∼20% of ciliary activity may be sufficient to prevent the development of rhinosinusitis. However, although administration of a β-galactosidase-expressing vector into control mice demonstrated efficient gene transfer to the nasal epithelium, treatment of Dnaic1−/− mice resulted in a low level of gene transfer, demonstrating that the severe rhinitis present in these animals impedes gene transfer. The results demonstrate that gene replacement therapy may be a viable treatment option for PCD, but further improvements in the efficiency of gene transfer are necessary.

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Acknowledgements

We thank Dr P Sears for technical assistance with the recording and analysis of ciliary activity for this project. This research was funded by NIH Grant 1RC1HL10081 to LEO and core grants NIH P30 DK065988, R026-CR11 from the Cystic Fibrosis Foundation, and IDG #2008-IDG-1017 from the NC Biotechnology Center. Research support was also provided by Oxford BioMedica to JCO.

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Authors and Affiliations

  1. Cystic Fibrosis and Pulmonary Diseases Research and Treatment Center, University of North Carolina, Chapel Hill, NC, USA

    L E Ostrowski, W Yin, M Patel, J Sechelski, T Rogers, K Burns, B R Grubb & J C Olsen

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  1. L E Ostrowski
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Correspondence to L E Ostrowski.

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Competing interests

Dr JCO is a named inventor on patents involving equine infectious anemia virus (EIAV)-based gene transfer technology and has received royalties for the use of EIAV-based vectors. The remaining authors declare no conflict of interest.

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Ostrowski, L., Yin, W., Patel, M. et al. Restoring ciliary function to differentiated primary ciliary dyskinesia cells with a lentiviral vector. Gene Ther 21, 253–261 (2014). https://doi.org/10.1038/gt.2013.79

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