四面体框架核酸对脑靶向肽分子的可控组装及性能  

Controllable Assembly and Properties of Brain Targeting Peptides with Tetrahedral Framework Nucleic Acids

作　　者：闫美玲 彭红珍[1,2] 左婷婷 田甜 诸颖[1,2] 孙艳红[1,2] YAN Mei-Ling;PENG Hong-Zhen;ZUO Ting-Ting;TIAN Tian;ZHU Ying;SUN Yan-Hong(CAS Key Laboratory of Interfacial Physics and Technology,Shanghai Institute of Applied Physics,Chinese Academy of Sciences,Shanghai 201800,China;The Interdiciplinary Research Center,Shanghai Advanced Research Institute,Chinese Academy of Sciences,Shanghai 201210,China;Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;University of Chinese Academy of Sciences,Beijing 100049,China)

机构地区：[1]中国科学院上海应用物理研究所,中国科学院微观界面物理与探测重点实验室,上海201800 [2]中国科学院上海高等研究院,基础交叉研究中心,上海201210 [3]上海中医药大学,上海201203 [4]中国科学院大学,北京100049

出　　处：《应用化学》2022年第10期1501-1509,共9页Chinese Journal of Applied Chemistry

基　　金：国家自然科学基金项目(Nos.22022410,81801818,62065012);上海市科委项目(Nos.19JC1410300,19142202400)资助。

摘　　要：通过设计带手臂链的四面体框架核酸,组装1~3条肽段修饰的手臂链的互补链,以及组装不同种类的脑靶向肽修饰的互补链,比较其毒性、细胞靶向摄取,探讨肽链的数量和种类对四面体DNA纳米探针脑靶向的影响。结果发现,在100 nmol/L浓度范围内,探针无细胞毒性。细胞在不同时间段内对脑靶向Angiopep-2(ANG)肽链修饰的四面体探针的摄取量与单纯四面体比较有显著性差异(P<0.01),在与脑微血管内皮细胞bEnd.3细胞共孵育0.5 h,与脑胶质瘤细胞U87共孵育1 h,四面体框架核酸组装3条肽链被摄取的量明显比组装1条肽链的摄取量多(P<0.01),但在与细胞共孵育2 h后,组装不同数量肽的探针在细胞内的摄取量没有显著差异。除此外,组装相同数量的噬菌体展示肽TGN和细胞穿膜肽TAT后,组装肽的四面体探针细胞摄取量比单纯四面体细胞内显著升高(P<0.01),但3种肽链组装的四面体探针在bEnd.3中细胞摄取量并无显著差异。这些结果表明,肽链的数量可能影响短时间内细胞对其摄取的速率,但并不影响2 h后细胞对探针的摄取总量,这一发现可有效节省出四面体修饰位点,为四面体框架核酸的更多功能修饰提供借鉴,在修饰单个靶向肽延长作用时间达到靶向目的的同时,可增加其它位点的功能化修饰,使其结构功能最大化,而且DNA框架核酸还可以作为其它脑靶向肽TGN和TAT的载体。In this study,the armed tetrahedral DNA nanostructures(TDN)are designed to bind single or three complementary strands of armed chain(1-ANG-TDN,3-ANG-TDN)which is modified with angiopep-2 or other brain targeting peptides(TAT,TGN).We compare the cell viability and cellular uptake of 1-ANGTDN or 3-ANG-TDN in brain capillary endothelial(bEnd.3)cells and Uppsala 87 malignant glioma(U87)cells.In addition,the cell viability and cellular uptake of ANG-TDN,TAT-TDN and TGN-TDN in bEnd.3 cells are investigated respectively.The results show that these probes are not cytotoxic to bEnd.3 cells and U87 cells at concentrationsbelow 100 nmol/L.After being incubated with bEnd.3cells for 0.5 h,the cellular uptake of 3-ANG-TDN is higher than that of 1-ANG-TDN(P<0.01).After being incubated with U87 cells for 1h,the cellular uptake of 3-ANG-TDN is higher than that of 1-ANG-TDN(P<0.01).After 2 h,there is no obvious difference in cellular uptake between 1-ANG-TDN and 3-ANG-TDN.ANG-TDN,TAT-TDN and TGN-TDN are up-taken by bEnd.3 cells much more than TDN(P<0.01).However,there isno statistical difference among ANG-TDN,TAT-TDN and TGN-TDN(P>0.05).These results indicate that the targeting of ANG-TDN probe can be improved by increasing the number of peptides or prolonging the co-incubation time.This discovery providesareference for the multifunctional modification of tetrahedral framework nucleic acid.When a single peptidelinked with TDN achieves the brain targeting,other sites of TDN can be modified by other functional groups to expand the structure and function.Tetrahedral framework nucleic acids can be used not only as an assembly platform for ANG peptides,but also for the assembly of other brain-targeting peptides(TAT and TGN)to design brain-targeting probes.

关 键 词：四面体框架核酸 脑靶向探针 可控组装 靶向摄取 

分 类 号：O655[理学—分析化学]