Abstract
Human endothelial cells are initiators and targets of the rejection response. Pre-operative modification of endothelial cells by small interfering RNA transfection could shape the nature of the host response post-transplantation. Ablation of endothelial cell class II major histocompatibility complex molecules by small interfering RNA targeting of class II transactivator can reduce the capacity of human endothelial cells to recruit and activate alloreactive T cells. Here, we report the development of small interfering RNA-releasing poly(amine-co-ester) nanoparticles, distinguished by their high content of a hydrophobic lactone. We show that a single transfection of small interfering RNA targeting class II transactivator attenuates major histocompatibility complex class II expression on endothelial cells for at least 4 to 6 weeks after transplantation into immunodeficient mouse hosts. Furthermore, silencing of major histocompatibility complex class II reduces allogeneic T-cell responses in vitro and in vivo. These data suggest that poly(amine-co-ester) nanoparticles, potentially administered during ex vivo normothermic machine perfusion of human organs, could be used to modify endothelial cells with a sustained effect after transplantation.
Highlights
Human endothelial cells are initiators and targets of the rejection response
As proof-of-principle, we demonstrate that human artery segments pretreated ex vivo using Small interfering RNA (siRNA)-loaded nanoparticles are refractory to major histocompatibility complex (MHC) class II molecule induction on endothelial cells for at least 4 to 6 weeks after transplantation into immunodeficient mouse hosts
Using fluorescent nanoparticles, loaded with DiD dye, and flow cytometry, we observed that PACE-50 nanoparticles associated strongly with human umbilical vein endothelial cells (HUVECs), whereas PACE-90 had significantly lower association (Fig. 2c). These results indicate that cellular toxicity and association vary with PDL content of the PACE nanoparticles, presumably due to differences in surface charge on the nanoparticles, which is due to the density of primary amines on the polymer chain
Summary
Human endothelial cells are initiators and targets of the rejection response. Pre-operative modification of endothelial cells by small interfering RNA transfection could shape the nature of the host response post-transplantation. We show that a single transfection of small interfering RNA targeting class II transactivator attenuates major histocompatibility complex class II expression on endothelial cells for at least 4 to 6 weeks after transplantation into immunodeficient mouse hosts. Silencing of major histocompatibility complex class II reduces allogeneic T-cell responses in vitro and in vivo These data suggest that poly(amine-co-ester) nanoparticles, potentially administered during ex vivo normothermic machine perfusion of human organs, could be used to modify endothelial cells with a sustained effect after transplantation. As proof-of-principle, we demonstrate that human artery segments pretreated ex vivo using siRNA-loaded nanoparticles are refractory to MHC class II molecule induction on endothelial cells for at least 4 to 6 weeks after transplantation into immunodeficient mouse hosts. Even with a single molecular target, this approach provides significant protection of human arterial grafts from rejection by adoptively transferred allogeneic T cells
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