Summary
The liver sinusoidal endothelial cells (SECs) possess unique hyaluronan receptors that recognize and internalize hyaluronic acid (HA). This characteristic was used in the development of a system for targeting foreign DNA to SECs. A gene carrier system was prepared by coupling HA (number-average molecular weight: 1.5 × 104) to poly-L-lysine (PLL, number-average molecular weight: 4.6 × 104) in a 1:1 weight ratio by reductive amination reaction. The resulting copolymer (PLL-g-HA) was mixed with various amounts of DNA in 154mM NaCl at 4°C. Inter-polyelectrolyte complex formation between PLL-g-HA and DNA exhibited minimal self-aggregation, explaining the highly soluble nature of the complex. The agarose gel retardation assay revealed that the titration point representing the minimum proportion of PLL-g-HA required to retard the DNA completely occurred at a 1:1 copolymer to DNA charge ratio. Intravenous injection of the [32P]pSV β-Gal plasmid complexed to PLL-g-HA in Wistar rats demonstrated specific hepatic targeting with >93% of the injected counts taken up by the liver in 1 h. Further, using a fluorescein isothiocyanate-labeled DNA, it was shown that the PLL-g-HA/DNA complex was distributed exclusively in the SECs. Seventy-two hours after injection of 90µ g of pSV β-Gal in a PLL-g-HA-complexed form, a large number of SECs expressing β-galactosidase were detected. Thus, the PLL-g-HA/DNA system permits targeted delivery of exogenous genes selectively to the liver SECs, providing a novel strategy for manipulation of SEC functions.
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© 2005 Springer-Verlag Tokyo
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Takei, Y., Ikejima, K., Enomoto, N., Maruyama, A., Sato, N. (2005). Genetic Manipulation of Liver Sinusoidal Endothelial Cells. In: Ishii, H., Suematsu, M., Tanishita, K., Suzuki, H. (eds) Organ Microcirculation. Keio University International Symposia for Life Sciences and Medicine, vol 13. Springer, Tokyo. https://doi.org/10.1007/4-431-27174-0_18
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DOI: https://doi.org/10.1007/4-431-27174-0_18
Publisher Name: Springer, Tokyo
Print ISBN: 978-4-431-22135-7
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