Effective mRNA Protection by Poly(l-ornithine) Synergizes with Endosomal Escape Functionality of a Charge-Conversion Polymer toward Maximizing mRNA Introduction Efficiency
Effective mRNA Protection by Poly(l-ornithine) Synergizes with Endosomal Escape Functionality of a Charge-Conversion Polymer toward Maximizing mRNA Introduction Efficiency
复制标题
DOI：
10.1002/marc.202100754
复制
发表时间：
2022-03-18
影响因子：
4.6
通讯作者：
Kataoka, Kazunori
中科院分区：
文献类型：
--
作者：
Dirisala, Anjaneyulu;Uchida, Satoshi;Kataoka, Kazunori
为了有效递送信使（m）RNA，递送载体需要两个主要功能：保护mRNA免受核酸酶的侵害和将mRNA从内溶酶体易位到细胞质。在本文中，这两种互补的功能性通过微调阳离子异构体化学结构和并入内体逃逸模态而整合到单个多聚物中。通过比较具有四亚甲基间隔基的聚（L-赖氨酸）（PLL）和具有三亚甲基间隔基的聚（L-鸟氨酸）（PLO）来评估亚甲基间隔基长度对阳离子聚合物侧链的影响。值得注意的是，mRNA/阳离子聚合物复合物的核酸酶稳定性在很大程度上受一个亚甲基基团的差异的影响，PLO/mRNA复合物与PLL/mRNA复合物相比显示出增强的稳定性。为了引入内体逃逸功能，PLO/mRNA复合物被电荷转换聚合物（CCP）包裹，其在细胞外pH下带负电荷，但在内体酸性pH下变成正电荷以破坏内体膜。与亲本PLO/mRNA复合物相比，CCP促进了培养细胞中复合物的内体逃逸，从而将mRNA的蛋白质表达效率提高了约80倍。总的来说，该系统协同PLO对核酸酶的保护作用和CCP在mRNA递送中的内体逃逸能力。
For efficient delivery of messenger (m)RNA, delivery carriers need two major functions: protecting mRNA from nucleases and translocating mRNA from endolysosomes to the cytoplasm. Herein, these two complementary functionalities are integrated into a single polyplex by fine-tuning the catiomer chemical structure and incorporating the endosomal escape modality. The effect of the methylene spacer length on the catiomer side chain is evaluated by comparing poly(l-lysine) (PLL) with a tetramethylene spacer and poly(L-ornithine) (PLO) with a trimethylene spacer. Noteworthily, the nuclease stability of the mRNA/catiomer polyplexes is largely affected by the difference in one methylene group, with PLO/mRNA polyplex showing enhanced stability compared to PLL/mRNA polyplex. To introduce the endosomal escape function, the PLO/mRNA polyplex is wrapped with a charge-conversion polymer (CCP), which is negatively charged at extracellular pH but turns positive at endosomal acidic pH to disrupt the endosomal membrane. Compared to the parent PLO/mRNA polyplex, CCP facilitated the endosomal escape of the polyplex in cultured cells to improve the protein expression efficiency from mRNA by approximately 80-fold. Collectively, this system synergizes the protective effect of PLO against nucleases and the endosomal escape capability of CCP in mRNA delivery.