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bioRxiv · 10.1101/2024.08.22.608643

PBAE-PEG based lipid nanoparticles for lung cell-specific gene delivery

Liu, B.·Sajiki, Y.·Sridharan, A.·Stuart, W.·Cui, X.·Siefert, M.·Araki, K.·Ziady, A.·Shi, D.·Whitsett, J. A.·Maeda, Y.
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Abstract

Delivery of modified mRNA encapsulated in lipid nanoparticles, exemplified by their successful use in COVID-19 vaccination, provides a framework for treating various genetic and acquired disorders. Herein, we developed PEGylated(PBAE-PEG) and non-PEGylated(PBAE) PBAE with lipids 4A3-SC8/DOPE/cholesterol/DOTAP to form lipid nanoparticles (LNPs) for mRNA delivery into different types of pulmonary cells in vivo. PBAE-PEG/LNP were highly active in transfecting HEK293T cells and air-liquid interfaced H441 cells in vitro. PBAE-PEG/LNP were used to express Cre-recombinase after administration to mice by intravenous injection, resulting in high transfection levels in pulmonary vascular endothelial cells. Intratracheal injection of both PBAE-PEG/LNP and PBAE/LNPs resulted in efficient and selective transfection of lung epithelial cells, identified by the expression of stabilized Cre-recombinase mRNA in club cells and alveolar type 2 cells. PBAE-PEG/LNP were most effective in transfecting alveolar type 2 cells after intratracheal injection, while PBAE/LNPs administered intratracheally were more effective in transfecting secretory airway cells. Cre-mediated recombination was specific to lung epithelial cells after intratracheal administration. Likewise, intravenous administration resulted in selective transfection of endothelial cells but not other pulmonary cell types, indicating their failure to cross the pulmonary endothelial-to-epithelial barrier. Moreover, 5-methoxyuridine modified mRNA was more efficient than unmodified mRNA in vivo but not in vitro.

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Liu, B., Sajiki, Y., Sridharan, A., Stuart, W., Cui, X., Siefert, M., Araki, K., Ziady, A., Shi, D., Whitsett, J. A., Maeda, Y.. 2024-08-23. PBAE-PEG based lipid nanoparticles for lung cell-specific gene delivery. https://doi.org/10.1101/2024.08.22.608643

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