Restoring ciliary function to differentiated primary ciliary dyskinesia cells with a lentiviral vector.
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 Dnaic1(flox/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.
Cystic Fibrosis and Pulmonary Diseases Research and Treatment Center, University of North Carolina, Chapel Hill, NC, USA.
L E
LE
Ostrowski
Cystic Fibrosis and Pulmonary Diseases Research and Treatment Center, University of North Carolina, Chapel Hill, NC, USA.
W
W
Yin
Cystic Fibrosis and Pulmonary Diseases Research and Treatment Center, University of North Carolina, Chapel Hill, NC, USA.
M
M
Patel
Cystic Fibrosis and Pulmonary Diseases Research and Treatment Center, University of North Carolina, Chapel Hill, NC, USA.
J
J
Sechelski
Cystic Fibrosis and Pulmonary Diseases Research and Treatment Center, University of North Carolina, Chapel Hill, NC, USA.
T
T
Rogers
Cystic Fibrosis and Pulmonary Diseases Research and Treatment Center, University of North Carolina, Chapel Hill, NC, USA.
K
K
Burns
Cystic Fibrosis and Pulmonary Diseases Research and Treatment Center, University of North Carolina, Chapel Hill, NC, USA.
B R
BR
Grubb
Cystic Fibrosis and Pulmonary Diseases Research and Treatment Center, University of North Carolina, Chapel Hill, NC, USA.
J C
JC
Olsen
D000818:Animals; D056444:Axonemal Dyneins / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D002923:Cilia / Q000378:metabolism / Q000502:physiology; D002925:Ciliary Motility Disorders / Q000628:therapy*; D004965:Estrogen Antagonists / Q000494:pharmacology; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics; D019426:Integrases / Q000235:genetics; D016086:Lentivirus / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D020545:Respiratory Mucosa / Q000187:drug effects / Q000378:metabolism; D012220:Rhinitis / Q000628:therapy; D012852:Sinusitis / Q000628:therapy; D013629:Tamoxifen / Q000494:pharmacology
D004965:Estrogen Antagonists; D013629:Tamoxifen; C045073:Cre recombinase; D019426:Integrases; D056444:Axonemal Dyneins; C550778:Dnaic1 protein, mouse