Chakraborty Samrat, Dlie Zewdu Yilma, Chakraborty Somdyuti, Roy Somdatta, Mukherjee Biswajit, Besra Shila Elizabeth, Dewanjee Saikat, Mukherjee Alankar, Ojha Probir Kumar, Kumar Vinay, Sen Ramkrishna
Pharmaceutics Research Laboratory, Department of Pharmaceutical Technology, Jadavpur University, Kolkata, India.
Pharmaceutics Research Laboratory, Department of Pharmaceutical Technology, Jadavpur University, Kolkata, India.
Mol Ther Nucleic Acids. 2020 Jun 5;20:34-49. doi: 10.1016/j.omtn.2020.01.034. Epub 2020 Feb 5.
Site-specific delivery of chemotherapeutics specifically to neoplastic hepatocytes without affecting normal hepatocytes should be a focus for potential therapeutic management of hepatocellular carcinoma (HCC). The aptamer TLS 9a with phosphorothioate backbone modifications (L5) has not been explored so far for preferential delivery of therapeutics in neoplastic hepatocytes to induce apoptosis. Thus, the objective of the present investigation was to compare the therapeutic potential of L5-functionalized drug nanocarrier (PTX-NPL5) with those of the other experimental drug nanocarriers functionalized by previously reported HCC cell-targeting aptamers and non-aptamer ligands, such as galactosamine and apotransferrin. A myriad of well-defined investigations such as cell cycle analysis, TUNEL (terminal deoxynucleotidyltransferase-mediated deoxyuridine triphosphate nick end labeling) assay, and studies related to apoptosis, histopathology, and immunoblotting substantiated that PTX-NPL5 had the highest potency among the different ligand-attached experimental formulations in inducing selective apoptosis in neoplastic hepatocytes via a mitochondrial-dependent apoptotic pathway. PTX-NPL5 did not produce any notable toxic effects in healthy hepatocytes, thus unveiling a new and a safer option in targeted therapy for HCC. Molecular modeling study identified two cell-surface biomarker proteins (tumor-associated glycoprotein 72 [TAG-72] and heat shock protein 70 [HSP70]) responsible for ligand-receptor interaction of L5 and preferential internalization of PTX-NPL5 via clathrin-mediated endocytosis in neoplastic hepatocytes. The potential of PTX-NPL5 has provided enough impetus for its rapid translation from the pre-clinical to clinical domain to establish itself as a targeted therapeutic to significantly prolong survival in HCC.
将化疗药物特异性地递送至肿瘤性肝细胞而不影响正常肝细胞，应成为肝细胞癌（HCC）潜在治疗管理的重点。具有硫代磷酸酯骨架修饰的适配体TLS 9a（L5），目前尚未探索其在肿瘤性肝细胞中优先递送治疗药物以诱导凋亡的作用。因此，本研究的目的是比较L5功能化药物纳米载体（PTX-NPL5）与其他实验性药物纳米载体的治疗潜力，后者由先前报道的HCC细胞靶向适配体和非适配体配体（如半乳糖胺和脱铁转铁蛋白）功能化。一系列明确的研究，如细胞周期分析、TUNEL（末端脱氧核苷酸转移酶介导的脱氧尿苷三磷酸缺口末端标记）测定以及与凋亡、组织病理学和免疫印迹相关的研究证实，PTX-NPL5在不同配体连接的实验制剂中，通过线粒体依赖性凋亡途径诱导肿瘤性肝细胞选择性凋亡的效力最高。PTX-NPL5在健康肝细胞中未产生任何明显的毒性作用，从而揭示了一种针对HCC的靶向治疗新的、更安全的选择。分子建模研究确定了两种细胞表面生物标志物蛋白（肿瘤相关糖蛋白72 [TAG-72]和热休克蛋白70 [HSP70]），它们负责L5的配体-受体相互作用以及PTX-NPL5在肿瘤性肝细胞中通过网格蛋白介导的内吞作用进行优先内化。PTX-NPL5的潜力为其从临床前快速转化到临床领域提供了足够的动力，使其成为一种靶向治疗药物，显著延长HCC患者的生存期。
