Transfer of the cystic fibrosis (CF) transmembrane conductance regulator (CFTR) to human CF cells mediated by extracellular vesicles
Authors
Cyrielle Vituret, Kathy Gallay, Marie-Pierre Confort, Najate Ftaich, Constantin I. Matei, Fabienne Archer, Corinne Ronfort, Jean-François Mornex, Marc Chanson, Attilio Di Pietro, Pierre Boulanger and Saw See Hong
Institution
University of Lyon
Country
France
Year
2016
Journal
Human Gene Therapy
Abstract
Cystic fibrosis (CF) is a genetic disease caused by mutations in the CF transmembrane
conductance regulator (CFTR) gene, resulting in a deficiency in chloride channel activity. In this study,
extracellular vesicles (EVs), microvesicles and exosomes, were used as vehicles to deliver exogenous
CFTR glycoprotein and its encoding mRNA (mRNAGFP-CFTR) to CF cells to correct the CFTR chloride channel
function. We isolated microvesicles and exosomes from the culture medium of CFTR-positive Calu-3
cells, or from A549 cells transduced with an adenoviral vector overexpressing a GFP-tagged CFTR (GFPCFTR).
Both microvesicles and exosomes had the capacity to package and deliver the GFP-CFTR
glycoprotein and mRNAGFP-CFTR to target cells in a dose-dependent manner. Homologous versus
heterologous EVs-to-cell transfer was studied, and it appeared that the cellular uptake of EVs was
significantly more efficient in homologous transfer. The incubation of CF15 cells, a nasal epithelial cell
line homozygous for the ΔF508 CFTR mutation, with microvesicles or exosomes loaded with GFP-CFTR
resulted in the correction of the CFTR function in CF cells in a dose-dependent manner. A time-course
analysis of EV-transduced CF cells suggested that CFTR transferred as mature glycoprotein was
responsible for the CFTR-associated channel activity detected at early times post-transduction, whereas
GFP-CFTR translated from exogenous mRNAGFP-CFTR was responsible for the CFTR function at later times.
Collectively, this study showed the potential application of microvesicles and exosomes as vectors for
CFTR transfer and functional correction of the genetic defect in human CF cells.