Magnetofection Enhances Adenoviral Vector-based Gene Delivery in Skeletal Muscle Cells.
The goal of magnetic field-assisted gene transfer is to enhance internalization of exogenous nucleic acids by association with magnetic nanoparticles (MNPs). This technique named magnetofection is particularly useful in difficult-to-transfect cells. It is well known that human, mouse, and rat skeletal muscle cells suffer a maturation-dependent loss of susceptibility to Recombinant Adenoviral vector (RAd) uptake. In postnatal, fully differentiated myofibers, the expression of the primary Coxsackie and Adenoviral membrane receptor (CAR) is severely downregulated representing a main hurdle for the use of these vectors in gene transfer/therapy. Here we demonstrate that assembling of Recombinant Adenoviral vectors with suitable iron oxide MNPs into magneto-adenovectors (RAd-MNP) and further exposure to a gradient magnetic field enables to efficiently overcome transduction resistance in skeletal muscle cells. Expression of Green Fluorescent Protein and Insulin-like Growth Factor 1 was significantly enhanced after magnetofection with RAd-MNPs complexes in C2C12 myotubes in vitro and mouse skeletal muscle in vivo when compared to transduction with naked virus. These results provide evidence that magnetofection, mainly due to its membrane-receptor independent mechanism, constitutes a simple and effective alternative to current methods for gene transfer into traditionally hard-to-transfect biological models.
Biochemistry Research Institute of La Plata (INIBIOLP)/National Scientific and Technical Research Council (CONICET), School of Medicine, National University of La Plata, La Plata, BA, Argentina (ZC 1900).
Andrea Soledad
AS
Pereyra
Ismaninger Street 22, Institute of Immunology and Experimental Klinikum rechts der Isar, Technical University of Munich, Munich, Germany (ZC 81675).
Olga
O
Mykhaylyk
Biochemistry Research Institute of La Plata (INIBIOLP)/National Scientific and Technical Research Council (CONICET), School of Medicine, National University of La Plata, La Plata, BA, Argentina (ZC 1900).
Eugenia Falomir
EF
Lockhart
Department of Internal Medicine, Section on Gerontology and Geriatric Medicine, Wake Forest School of Medicine, Winston-Salem, NC, USA (ZC 27157).
Jackson Richard
JR
Taylor
Department of Internal Medicine, Section on Gerontology and Geriatric Medicine, Wake Forest School of Medicine, Winston-Salem, NC, USA (ZC 27157).
Osvaldo
O
Delbono
Biochemistry Research Institute of La Plata (INIBIOLP)/National Scientific and Technical Research Council (CONICET), School of Medicine, National University of La Plata, La Plata, BA, Argentina (ZC 1900).
Rodolfo Gustavo
RG
Goya
Ismaninger Street 22, Institute of Immunology and Experimental Klinikum rechts der Isar, Technical University of Munich, Munich, Germany (ZC 81675).
Christian
C
Plank
Biochemistry Research Institute of La Plata (INIBIOLP)/National Scientific and Technical Research Council (CONICET), School of Medicine, National University of La Plata, La Plata, BA, Argentina (ZC 1900); IFEC-CONICET, Farmacology Department, School of Chemistry, National University of Cordoba, (ZC 5000) Córdoba, Argentina.
Claudia Beatriz
CB
Hereñu
Adenoviral vectors; Gene delivery; Magnetic nanoparticles; Magneto-adenovectors; Magnetofection; Skeletal muscle