Institute of Nano Science and Technology, Sector-81, Knowledge City, Sahibzada Ajit Singh Nagar, Punjab 140306, India.
Department of Pharmacology, Chandigarh College of Pharmacy, Sector 112, Sahibzada Ajit Singh Nagar, Punjab 140307, India.
ACS Appl Bio Mater. 2021 Jun 21;4(6):5324-5335. doi: 10.1021/acsabm.1c00437. Epub 2021 Jun 3.
We report the formulation of aminocellulose-grafted polymeric nanoparticles containing LCS-1 for synthetic lethal targeting of checkpoint kinase 2 (CHEK2)-deficient HCT116 colon cancer (CRC) cells to surpass the limitations associated with the solubility of LCS-1 (a superoxide dismutase inhibitor). Aminocellulose (AC), a highly biocompatible and biodegradable hydrophilic polymer, was grafted over polycaprolactone (PCL), and a nanoprecipitation method was employed for formulating nanoparticles containing LCS-1. In this study, we exploited the synthetic lethal interaction between SOD1 and CHEK2 for the specific inhibition of CHEK2-deficient HCT116 CRC cells using LCS-1-loaded PCL-AC NPs. Furthermore, the effects of formation of protein corona on PCL-AC nanoparticles were also assessed in terms of size, cellular uptake, and cell viability. LCS-1-loaded NPs were evaluated for their size, zeta potential, and polydispersity index using a zetasizer, and their morphological characteristics were assessed by transmission electron microscopy, scanning electron microscopy, and atomic force microscopy analyses. Cellular internalization using confocal microscopy exhibited that nanoparticles were uptaken by HCT116 cells. Also, nanoparticles were cytocompatible as they did not induce cytotoxicity in hTERT and HEK-293 cells. The LCS-1-loaded PCL-AC NPs were quite hemocompatible and were 240 times more selective in killing CHEK2-deficient cells as compared to CHEK2-proficient CRC cells. Moreover, PCL-AC NPs exhibited that the protein corona-coated nanoparticles were incubated in the human and fetal bovine sera as visualized by SDS-PAGE. A slight increment in hydrodynamic diameter was observed for corona-coated PCL-AC nanoparticles, and size increment was further confirmed by TEM. Corona-coated PCL-AC NPs also exhibited cellular uptake as demonstrated by flow cytometric analysis and did not cause cytotoxic effects on hTERT cells. The nanoformulation was developed to enhance therapeutic potential of the drug LCS-1 for enhanced lethality of colorectal cancer cells with CHEK2 deficiency.
我们报告了载有 LCS-1 的氨基纤维素接枝聚合物纳米粒子的配方，用于合成致死靶向 checkpoint kinase 2（CHEK2）缺陷的 HCT116 结肠癌细胞（CRC），以克服与 LCS-1（超氧化物歧化酶抑制剂）的溶解度相关的限制。氨基纤维素（AC）是一种高度生物相容和可生物降解的亲水性聚合物，接枝在聚己内酯（PCL）上，并采用纳米沉淀法制备载有 LCS-1 的纳米粒子。在这项研究中，我们利用 SOD1 和 CHEK2 之间的合成致死相互作用，使用载有 LCS-1 的 PCL-AC NPs 特异性抑制 CHEK2 缺陷的 HCT116 CRC 细胞。此外，还评估了形成蛋白冠对 PCL-AC 纳米粒子的大小、细胞摄取和细胞活力的影响。使用 Zetasizer 评估载有 LCS-1 的 NPs 的粒径、Zeta 电位和多分散指数，并通过透射电子显微镜、扫描电子显微镜和原子力显微镜分析评估其形态特征。使用共聚焦显微镜进行细胞内摄取显示，纳米粒子被 HCT116 细胞摄取。此外，纳米粒子具有细胞相容性，因为它们不会在 hTERT 和 HEK-293 细胞中引起细胞毒性。载有 LCS-1 的 PCL-AC NPs 具有良好的血液相容性，与 CHEK2 功能正常的 CRC 细胞相比，对 CHEK2 缺陷细胞的选择性杀伤高 240 倍。此外，PCL-AC NPs 表明，在人血清和胎牛血清中孵育蛋白冠包被的纳米粒子时，如 SDS-PAGE 所示。观察到包被蛋白冠的 PCL-AC 纳米粒子的水动力直径略有增加，并且 TEM 进一步证实了尺寸的增加。包被蛋白冠的 PCL-AC NPs 也表现出细胞摄取，如通过流式细胞术分析所示，并且对 hTERT 细胞没有引起细胞毒性作用。该纳米制剂的开发旨在增强药物 LCS-1 的治疗潜力，以增强 CHEK2 缺陷的结直肠癌细胞的致死性。
