mRNA loading into ATP-responsive polyplex micelles with optimal density of phenylboronate ester crosslinking to balance robustness in the biological milieu and intracellular translational efficiency
mRNA loading into ATP-responsive polyplex micelles with optimal density of phenylboronate ester crosslinking to balance robustness in the biological milieu and intracellular translational efficiency
复制标题
DOI：
10.1016/j.jconrel.2020.12.033
复制
发表时间：
2021-02-10
影响因子：
10.8
通讯作者：
Kataoka, Kazunori
中科院分区：
文献类型：
--
作者：
Yoshinaga, Naoto;Uchida, Satoshi;Kataoka, Kazunori
信使RNA的载体需要保护信使RNA不被酶降解，并选择性地在胞浆中释放信使RNA，以使信使RNA顺利翻译。为了满足这些要求，我们通过将信使核糖核酸与苯基硼酸衍生的聚乙二醇阳离子嵌段共聚物和多元醇基络合，通过苯基硼酸酯的自发形成形成交联结构，设计了内核具有三磷酸腺苷响应性交联的信使核糖核酸聚合物胶束(PM)。这样制备的PMS对酶攻击是耐受的，当响应于升高的ATP浓度而被苯基硼酸酯连接断裂触发时，PMS反过来在胞浆中解体以释放mRNA。PM的两个结构因素，包括(I)苯基硼酸酯交联剂的引入比例和(Ii)聚阳离子片段中氨基的结构和质子化程度，对于最大限度地提高培养细胞中蛋白质的表达至关重要，因为在生物环境中的稳健性和胞浆中ATP响应的mRNA释放之间取得了优化的平衡。通过在嵌段共聚物的mRNA和omega-end中安装胆固醇部分，进一步稳定了最佳PM配方，以延长静脉注射后的血液循环寿命。
Carriers for messenger RNA (mRNA) delivery require propensities to protect the mRNA from enzymatic degradation and to selectively release mRNA in the cytosol for smooth mRNA translation. To meet these requirements, we designed mRNA-loaded polyplex micelles (PMs) with ATP-responsive crosslinking in the inner core by complexing mRNA with poly(ethylene glycol)-polycation block copolymers derivatized with phenylboronic acid and polyol groups, which form crosslinking structures via spontaneous phenylboronate ester formation. PMs thus prepared are tolerable against enzymatic attack and, in turn, disintegrate in the cytosol to release mRNA when triggered by the cleavage of phenylboronate ester linkages in response to elevated ATP concentration. Two structural factors of the PM, including (i) the introduction ratios of phenylboronate ester crosslinkers and (ii) the structure and protonation degree of amino groups in the polycation segment, are critical for maximizing protein expression in cultured cells due to the optimized balance between the robustness in the biological milieu and the ATP-responsive mRNA release in the cytosol. The optimal PM formulation was further stabilized by installing cholesterol moieties into both the mRNA and omega-end of the block copolymer to elicit longevity in blood circulation after intravenous injection.