Abstract

Nanoparticles composed of poly(alkyl cyanoacrylate) (PACA) have shown great promise due to their biodegradability and high drug loading capacity. Development of optimal PACA nanocarriers requires detailed analysis of the overall cellular impact exerted by PACA variants. We here perform a comprehensive comparison of cabazitaxel (CBZ)-loaded nanocarriers composed of three different PACA monomers, i.e. poly(n-butyl cyanoacrylate) (PBCA), poly(2-ethylbutyl cyanoacrylate) (PEBCA) and poly(octyl cyanoacrylate) (POCA). The cytotoxicity of drug-loaded and empty PACA nanoparticles were compared to that of free CBZ across a panel of nine cancer cell lines by assessing cellular metabolism, proliferation and protein synthesis. The analyses revealed that the cytotoxicity of all CBZ-loaded PACAs was similar to that of free CBZ for all cell lines tested, whereas the empty PACAs exerted much lower toxicity. To increase our understanding of the toxic effects of these treatments comprehensive MS-based proteomics were performed with HCT116, MDA-MB-231 and PC3 cells incubated with PACA-CBZ variants or free CBZ. Interestingly, PACA-CBZ specifically led to decreased levels of proteins involved in focal adhesion and stress fibers in all cell lines. Since we recently demonstrated that encapsulation of CBZ within PEBCA nanoparticles significantly improved the therapeutic effect of CBZ on a patient derived xenograft model in mice, we investigated the effects of this PACA variant more closely by immunoblotting. Interestingly, we detected several changes in the protein expression and degree of phosphorylation of SRC-pathway proteins that can be relevant for the therapeutic effects of these substances.

摘要

由聚（氰基丙烯酸烷基酯）（PACA）组成的纳米颗粒由于其生物降解性和高载药量而显示出巨大的前景。开发最佳 PACA 纳米载体需要详细分析 PACA 变体对细胞的整体影响。我们在这里对由三种不同的 PACA 单体组成的卡巴他赛 (CBZ) 负载纳米载体进行了全面比较，即聚（氰基丙烯酸正丁酯）（PBCA）、聚（2-乙基氰基丙烯酸酯）（PEBCA）和聚（氰基丙烯酸辛酯）（ POCA）。通过评估细胞代谢、增殖和蛋白质合成，将载药和空的 PACA 纳米粒子的细胞毒性与游离 CBZ 的细胞毒性在九个癌细胞系中进行了比较。分析表明，对于所有测试的细胞系，所有负载 CBZ 的 PACA 的细胞毒性与游离 CBZ 的细胞毒性相似，而空的 PACA 的毒性要低得多。为了增加我们对这些治疗的毒性作用的理解，我们对 HCT116、MDA-MB-231 和 PC3 细胞与 PACA-CBZ 变体或游离 CBZ 一起孵育进行了基于 MS 的综合蛋白质组学。有趣的是，PACA-CBZ 特别导致所有细胞系中参与粘着斑和应力纤维的蛋白质水平降低。由于我们最近证明将 CBZ 封装在 PEBCA 纳米颗粒中显着提高了 CBZ 对小鼠异种移植模型的治疗效果，因此我们通过免疫印迹更密切地研究了这种 PACA 变体的影响。有趣的是，我们检测到 SRC 通路蛋白的蛋白质表达和磷酸化程度的一些变化，这些变化可能与这些物质的治疗效果有关。