Generation of magnetic nanoparticle-adenovector complexes and their use for magnetic field-assisted transduction of cell lines.

SCHWERDT J; HEREÑU C; CAMINOA C; MYKHAYLYK O; GOYA R

Mar del Plata

Congreso; ) LV Reunión Científica Anual de la Sociedad Argentina de Investigación Clínica- SAIC; 2010

The implementation of neuroprotective gene therapy for Parkinson´s and Alzheimer´s disease is a promising approach but up to now gene delivery to the brain requires direct injection of gene vectors into the target brain region. The association of viral vector-based gene delivery with nanotechnology now offers the possibility of developing minimally invasive gene therapy strategies for the brain. In order to explore this alternative we have constructed two adenoviral vectors, RAd-DsRed2 which expresses DsRed2, a red fluorescent protein from Discosoma, and RAd-GFP a vector expressing green fluorescent protein (GFP). The vectors were constructed by the two-plasmid method. These vectors were or are to be complexed with three types of magnetic nanoparticles (MNP) whose physical characteristics are shown in Table 1. Non complexed vectors were tested at varying multiplicities of infection (MOI) in different cell lines (HEK 293, B92, N2a and muscle cells C2C12 and showed pancellular expression of the corresponding reporter gene. In the rat brain, the vectors showed appropriate levels of expression in the hypothalamus, susbtantia nigra and the ependymal cell layer. Subsequently, MNP-RAd complexes were generated and tested on the above cell lines under the influence of an external magnetic field. Magnetic field-assisted reporter gene delivery (magnetotransduction) led to a MNP-dose dependent enhancement of gene expression. We conclude that our MNP-RAd complexes are suitable for magnetotransduction in vitro. These magnetic adenovectors are to be used for future in vivo magnetotransduction studies in the rat brain.