苯硼酸修饰的聚乙烯亚胺系统在 T 细胞小 RNA 转染的应用

小 RNA 是 T 细胞发育、分化和功能的重要调控分子，但目前仍缺乏安全有效的 T 细胞 RNA 转 染系统。基于 T 细胞表面唾液酸化的特性，该文利用苯硼酸和唾液酸的相互作用，带动苯硼酸修饰的聚乙烯亚胺系统(Polyethylenimine-Phenylboronic Acid，PEI-PBA)介导小 RNA 的 T 细胞转染。通过细胞计数试剂盒(CCK-8)和羟基荧光素二醋酸盐琥珀酰亚胺脂(CFSE)实验显示，PEI-PBA 转染系统对 T 细胞无显著细胞毒性和异常增殖。同时，流式检测人源抗 CD3/28 磁珠激活下的分化抗原 3(CD3)阳性 T 细胞在 PEI-PBA 转染系统下，小 RNA 平均摄取率增加到18.43%，而在鼠源 T 细胞中 PEI-PBA 介导的小 RNA 转染并无明显效果。结果显示，PEI-PBA 纳米转染系统介导小 RNA 的递送具有无毒高效的特点。

Delivery of Small RNAs by Phenylboronic Acid-Grafted Polyethylenimine Nanoparticles into T Lymphocytes

Small RNAs are essential regulators for T cell development, differentiation and functions. However, it is hard to deliver small RNAs into primary T cells by conventional transfection methods. Here, we reported that amphiphilic PBA-grafted PEI1.8k (PEI-PBA) nanovector facilitates the primary T cell-targeted RNA delivery miRNAthrough recognition of sialic groups on cell membrane of the T lymphocytes. CCK-8 (cell counting kit-8) and CFSE (5,6-carboxyfluorescein diacetate, succinimidyl ester) assay showed that the administration of PEI-PBA did not cause significant cell toxicity or abnormal proliferation. Meanwhile, the flowcytometry showed the average intake of miRNA was increased to 18.43% in the anti-CD3/CD28 activated CD3+ (cluster of differentiation 3) T lymphocytes by PEI-PBA in human, but not mouse. The results showed that PEI-PBA nano-system had effective delivery of small RNAs in human primary T cells without toxcity.