Transfert d'ARNm par des lipopolyplexes et vaccination antimélanome : ciblage des cellules dendritiques à l'aide de lipopolyplexes mannosylés

Abstract : Previously, it has been demonstrated that mice vaccination with lipopolyplexes (LPR) containing melanoma antigen MART1 mRNA can induce the generation of specific cytotoxic T cells and delay B16F10 melanoma growth and lung metastases. LPR are ternary complexes consisting of mRNA, a histidylated cationic polymer and histidylated cationic liposomes. The objective of my thesis was to enhance this antitumor vaccination through the development of new liposomes that can target specifically dendritic cells (DC). The targeting of DC was achieved by incorporating a mannosylated glycolipid within liposomes to enhance their recognition by the mannose receptor. From these liposomes, formulations based ternary complexes of DNA (mannosylated-LPD or Man11-LPD100) or formulations based on mRNA (mannosylated LPR or Man11 LPR100) were developed. The results show that formulations made with Man11-LPD100 are better internalized and transfect efficiently DC than LPD100. Man11 LPR100 transfect with greater efficiency DC compared to DNA based formulation (Man11-LPD100). Furthermore, a strong reduction of the toxicity of LPD was obtained by liposomes dialysis. It is also possible to preserve their activity by freeze-drying. Mice scintigraphy revealed that 9% of LPD are captured in the spleen following IV injection. We demonstrated after isolation of splenic DC that Man11-LPR100 transfect DC 4 times more than LPR100. Finally, immunization of mice with Man11-LPR100 containing mRNA MART1 allows a more effective vaccination against B16F10 tumor and a better mice survival than non-mannosylated ones. In conclusion, Man11-LPR100 are promising vectors to target and transfect splenic DC with a tumor antigen mRNA aiming to an induction of an immune response against tumor cells.
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Federico Perche. Transfert d'ARNm par des lipopolyplexes et vaccination antimélanome : ciblage des cellules dendritiques à l'aide de lipopolyplexes mannosylés. Médecine humaine et pathologie. Université d'Orléans, 2010. Français. ⟨NNT : 2010ORLE2068⟩. ⟨tel-00665118⟩

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