Glioma Dual-Targeting Nanohybrid Protein Toxin Constructed by Intein-Mediated Site-Specific Ligation for Multistage Booster Delivery.
Glioma Dual-Targeting Nanohybrid Protein Toxin Constructed by Intein-Mediated Site-Specific Ligation for Multistage Booster Delivery.
复制标题
通过内含肽介导的位点特异性连接构建神经胶质瘤双靶向纳米混合蛋白毒素，用于多级加强递送
DOI：
10.7150/thno.20578
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发表时间：
2017
期刊：
影响因子：
12.4
通讯作者：
Huang Y
中科院分区：
文献类型：
--
作者：
Chen Y;Zhang M;Jin H;Li D;Xu F;Wu A;Wang J;Huang Y
恶性神经胶质瘤是最难以治愈的癌症之一，因为它具有强大的血脑屏障（BBB），很少有治疗方法可以穿透并到达肿瘤。在过去的二十年里，生物制剂在癌症治疗中得到了蓬勃发展，但由于血脑屏障的不渗透性，生物制剂在脑肿瘤中的应用一直被忽视。事实上，大分子药物的脑传递是一个长期未解决的问题，它已成为医学和制药科学的圣杯。即使通过靶向配体来辅助，由于配体修饰蛋白质的合成困难，蛋白质脑递送仍然具有挑战性。在此，我们提出了一种类似火箭的多级助推器，用于抗胶质瘤治疗的蛋白毒素，trichosanthin （TCS）。TCS是一种核糖体失活蛋白，对多种实体肿瘤具有较强的活性，但缺乏特异性作用和细胞穿透能力。为了克服其耐药性差和位点特异性修饰的挑战，采用内部介导的连接方法，将明胶酶可切割肽和细胞穿透肽（CPP）融合的重组TCS毒素位点特异性地偶联到乳铁蛋白（LF）上，从而构建了一个穿透bbb、明胶酶可激活的细胞穿透纳米杂化TCS毒素。这种纳米混合TCS系统的特点是多级增强策略，用于胶质瘤双靶向递送。首先，LF可以靶向过表达血脑屏障的低密度脂蛋白受体相关蛋白-1 (LRP-1)，并协助血脑屏障渗透。其次，一旦到达肿瘤部位，明胶酶可切割肽作为一种分离器响应胶质瘤相关基质金属蛋白酶（MMPs），从而释放到pcp融合的毒素。第三，CPP介导TCS毒素在肿瘤内和细胞内的渗透，从而增强其抗肿瘤活性。证实了该递送系统的血脑屏障穿透性和mmp -2活化性。在皮下和原位动物模型中评估其抗胶质瘤活性。我们的工作为提高这类蛋白毒素的可药物性，促进其在体内靶向癌症治疗中的应用提供了一个有用的方案。
Malignant glioma is one of the most untreatable cancers because of the formidable blood-brain barrier (BBB), through which few therapeutics can penetrate and reach the tumors. Biologics have been booming in cancer therapy in the past two decades, but their application in brain tumor has long been ignored due to the impermeable nature of BBB against effective delivery of biologics. Indeed, it is a long unsolved problem for brain delivery of macromolecular drugs, which becomes the Holy Grail in medical and pharmaceutical sciences. Even assisting by targeting ligands, protein brain delivery still remains challenging because of the synthesis difficulties of ligand-modified proteins. Herein, we propose a rocket-like, multistage booster delivery system of a protein toxin, trichosanthin (TCS), for antiglioma treatment. TCS is a ribosome-inactivating protein with the potent activity against various solid tumors but lack of specific action and cell penetration ability. To overcome the challenge of its poor druggability and site-specific modification, intein-mediated ligation was applied, by which a gelatinase-cleavable peptide and cell-penetrating peptide (CPP)-fused recombinant TCS toxin can be site-specifically conjugated to lactoferrin (LF), thus constructing a BBB-penetrating, gelatinase-activatable cell-penetrating nanohybrid TCS toxin. This nanohybrid TCS system is featured by the multistage booster strategy for glioma dual-targeting delivery. First, LF can target to the BBB-overexpressing low-density lipoprotein receptor-related protein-1 (LRP-1), and assist with BBB penetration. Second, once reaching the tumor site, the gelatinase-cleavable peptide acts as a separator responsive to the glioma-associated matrix metalloproteinases (MMPs), thus releasing to the CPP-fused toxin. Third, CPP mediates intratumoral and intracellular penetration of TCS toxin, thereby enhancing its antitumor activity. The BBB penetration and MMP-2-activability of this delivery system were demonstrated. The antiglioma activity was evaluated in the subcutaneous and orthotopic animal models. Our work provides a useful protocol for improving the druggability of such class of protein toxins and promoting their in-vivo application for targeted cancer therapy.