Targeted gene delivery to sinusoidal endothelial cells: DNA nanoassociate bearing hyaluronan‐glycocalyx
Targeted gene delivery to sinusoidal endothelial cells: DNA nanoassociate bearing hyaluronan‐glycocalyx
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DOI：
10.1096/fj.03-0494fje
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发表时间：
2004-04
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通讯作者：
Y. Takei;A. Maruyama;A. Ferdous;Y. Nishimura;S. Kawano;K. Ikejima;Shigetoshi Okumura;S. Asayama;M. Nogawa;M. Hashimoto;Y. Makino;Masahiko Kinoshita;Sumio Watanabe;T. Akaike;J. Lemasters;N. Sato
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作者：
Y. Takei;A. Maruyama;A. Ferdous;Y. Nishimura;S. Kawano;K. Ikejima;Shigetoshi Okumura;S. Asayama;M. Nogawa;M. Hashimoto;Y. Makino;Masahiko Kinoshita;Sumio Watanabe;T. Akaike;J. Lemasters;N. Sato
肝窦内皮细胞(SECs)具有识别和内化透明质酸(HA)的独特受体。为了建立一种将外源DNA靶向SECS的体系，通过将HA偶联到聚L赖氨酸(PLL)上，制备了含有HA侧链的梳状聚阳离子。将得到的HA接枝-PLL共聚物(PLL-g-HA)与DNA在154 mM的氯化钠溶液中混合，形成含有水化透明质酸盐壳的可溶纳米缔合物。琼脂糖凝胶滞留分析表明，尽管存在多阴离子HA侧链，但PLL骨架与DNA有选择性的相互作用。为了确定PLL-g-HA/DNA复合体在体内是否被SEC HA受体识别，我们给Wistar大鼠静脉注射。经尾静脉给药，将磷脂酶-g-HA与~(32)P标记的β-半乳糖苷酶表达载体(pSV-β-Gal)复合。注射后1小时，90%的放射性仍留在肝脏。给药的PLL-g-HA与FITC标记的DNA的复合体显示，载体-DNA复合体仅分布在SEC中。转染β-g-HA/PSVβ-Gal后，在肝窦内壁可检测到大量表达PLL-g-HA/PSV-Gal的SECs。此外，PLL-g-HA有效地稳定了DNA三链的形成。总之，新的PLL-g-HA/DNA载体系统允许将外源基因选择性地靶向转移到SEC。
Liver sinusoidal endothelial cells (SECs) possess unique receptors that recognize and internalize hyaluronic acid (HA). To develop a system for targeting foreign DNA to SECs, comb‐type polycations having HA side chains were prepared by coupling HA to poly(L‐lysine) (PLL). The HA‐grafted‐PLL copolymer (PLL‐g‐HA) thus formed was mixed with DNA in 154 mM NaCl to form soluble nanoassociates bearing hydrated hyaluronate shells. Agarose gel retardation assays revealed selective interaction of the PLL backbone with DNA despite the presence of polyanionic HA side chains. To determine whether the PLL‐g‐HA/DNA complexes were recognized by SEC HA receptors in vivo, we injected Wistar rats i.v. via the tail vein with PLL‐ g‐HA complexed to a β‐galactosidase expression plasmid (pSV β‐Gal) labeled with 32P. One hour postinjection, >90% of the injected radioactivity remained in the liver. Administration of the PLL‐g‐HA complexed to an FITC‐labeled DNA revealed that the carrier‐DNA complex was distributed exclusively in SECs. A large number of SECs expressing β‐galactosidase was detected along the sinusoidal lining after transfection with PLL‐g‐HA/pSV β‐Gal. Moreover, PLL‐g‐HA effectively stabilized DNA triplex formation. In conclusion, the new PLL‐g‐ HA/DNA carrier system permits targeted transfer of exogenous genes selectively to the SECs.