monopDNA Virus Inspired NanoParticles for gene transfer – VINP

Gene transfer is a new therapeutic opportunity to cure disease without efficient treatment. Most of the tries today rely on viral carrier, with limitations that needs to be circumvented. An alternative to the use of viruses is the synthetic carriers. However, the gene transfer efficiency of synthetic gene carriers remains low, to be used in large scale and benefit from a large economical development. Viruses use the cell machinery to deliver genetic information inside the nucleus in cells of a targeted organ. Using the tremendous adaptation of viruses to the cell machinery, we suggest building polymer-based nanoparticles that will take advantage of the cell machinery to improve the transportation of nucleic acids. The nanoparticles will be plurifunctional, in order to alleviate the extra and intracellular barriers. We suggest using a new cationic polymer-pDNA construction to build nanoparticle containing only one pDNA copy (the ultimate small particle size), mimicking to some extent the viruses such r-AAV. A new formulation process will provide the formation of core-shell nanoparticles with a pDNA copy embedded in stealth corona allowing the extracellular targeting, a fast take-up by the cell using the clathrin-mediated endosomal mechanism, then a fast endosomal release of the nanoparticle before the nanoparticle disassembling in the cytosol. The functional pDNA will then ensure a fast diffusion toward the nucleus, and an improved nuclear internalization. The building and internal organization of the nanoparticles will be probed thanks to scattering experiments (SANS and SAXS, time-resoled), and the relevance of the various suggested functions borne by the cationic polymer or the pDNA will be demonstrated, by themselves first, and then integrated in the nanoparticles. Gene transfer efficiency will then be evaluated in vitro, then in vivo as a function of the administration mode, systemic for the hepatocyte-targeting nanoparticles and aerosolization for the lung epithelial cells-targeting nanoparticles. Mechanisms of cell trafficking will be probed, and correlated to the various structures/functions of the nanoparticles. Biomarkers analysis will be conducted during in vivo testing in order to evaluate the tolerance of these nanoparticles, as a function of the administration mode.