Wang Zhao, Sun Jingjing, Jia Lin, Sheng Ruilong
School of Material Engineering, Jinling Institute of Technology, Nanjing, 211169, China.
Department of Pharmaceutical Sciences, College of Pharmacy, University of Nebraska Medical Center, Omaha, NE, USA.
J Mater Sci Mater Med. 2025 Jun 6;36(1):47. doi: 10.1007/s10856-024-06837-4.
Glycopolymer drug delivery nanosystems have attracted increasing attention in the field of sustainable biomaterials and clinical biomedicine, while few studies addressed their intracellular drug delivery properties, endocytosis pathways, intracellular trafficking, autophagy behaviors and the effect of autophagy inhibitors. Based on our previous study, in this work, a pH-responsive glycopolymer (PMAgala-b-P(MAA-co-MAChol)) was synthesized and used as a drug delivery carrier, to encapsulate antitumor drug doxorubicin (DOX) into nanomicelles, with high DOX loading efficiency and pH-responsive DOX release properties. The cytotoxicity, cell proliferation inhibition, endocytosis pathway, intracellular trafficking/localization, and autophagy behavior of the blank glycopolymer micelles and/or DOX-loaded micelles were studied in a Human Glioblastoma Carcinoma (H4) and green fluorescent protein-tagged H4-GFP-LC3 cell lines. The glycopolymer micelles could be taken up into the cells through favorable caveolae-mediated and clathrin-mediated endocytic pathways, and their intracellular trafficking/localization were associated with endosome-lysosome systems. Notably, after treating with DOX-loaded glycopolymer micelles (or free DOX) to the H4-GFP-LC3 cells, exogenous substances-induced autophagosome accumulation was observed. The autophagy inhibitors: 3-methyladenine (3-MA) and hydroxychloroquine (HCQ) were used to monitor the autophagy behavior of H4-GFP-LC3 cells incubated with the micelles. Interestingly, the autophagy inhibitors could significantly enhance the antitumor performance of the free DOX and/or DOX-loaded micelles, the drug combination effect of autophagy inhibitors and DOX was further studied by Bliss independent model analysis. Taken together, this work provided a preliminary understanding of the intracellular drug delivery properties of glycopolymer micelles and demonstrated the effect of different autophagy inhibitors, which might inspire future innovation of "autophagy regulator-combined nanotherapeutics" toward efficient cancer chemotherapy.
糖聚合物药物递送纳米系统在可持续生物材料和临床生物医学领域引起了越来越多的关注，然而很少有研究涉及其细胞内药物递送特性、内吞途径、细胞内运输、自噬行为以及自噬抑制剂的作用。基于我们之前的研究，在这项工作中，合成了一种pH响应性糖聚合物（PMAgala-b-P(MAA-co-MAChol)）并用作药物递送载体，将抗肿瘤药物阿霉素（DOX）封装到纳米胶束中，具有高DOX负载效率和pH响应性DOX释放特性。在人胶质母细胞瘤癌（H4）和绿色荧光蛋白标记的H4-GFP-LC3细胞系中研究了空白糖聚合物胶束和/或载DOX胶束的细胞毒性、细胞增殖抑制、内吞途径、细胞内运输/定位和自噬行为。糖聚合物胶束可以通过有利的小窝介导和网格蛋白介导的内吞途径被细胞摄取，并且它们的细胞内运输/定位与内体-溶酶体系统相关。值得注意的是，用载DOX的糖聚合物胶束（或游离DOX）处理H4-GFP-LC3细胞后，观察到外源性物质诱导的自噬体积累。使用自噬抑制剂：3-甲基腺嘌呤（3-MA）和羟氯喹（HCQ）来监测与胶束孵育的H4-GFP-LC3细胞的自噬行为。有趣的是，自噬抑制剂可以显著增强游离DOX和/或载DOX胶束的抗肿瘤性能，通过Bliss独立模型分析进一步研究了自噬抑制剂和DOX的药物联合作用。综上所述，这项工作提供了对糖聚合物胶束细胞内药物递送特性的初步理解，并证明了不同自噬抑制剂的作用，这可能会激发未来“自噬调节剂联合纳米疗法”对高效癌症化疗的创新。
