Enhancement of tumor uptake and therapeutic efficacy of EGFR-targeted antibody cetuximab and antibody-drug conjugates by cholesterol sequestration
Enhancement of tumor uptake and therapeutic efficacy of EGFR-targeted antibody cetuximab and antibody-drug conjugates by cholesterol sequestration
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通过胆固醇封存增强 EGFR 靶向抗体西妥昔单抗和抗体药物缀合物的肿瘤摄取和治疗功效。
DOI：
10.1002/ijc.28950
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发表时间：
2015-01-01
影响因子：
6.4
通讯作者：
Luo, Yongzhang
中科院分区：
文献类型：
--
作者：
Chen, Yang;Liu, Guanghua;Luo, Yongzhang
西妥昔单抗是一种针对表皮生长因子受体 (EGFR) 的单克隆抗体 (mAb)，作为一种有前途的癌症治疗策略已得到深入研究。西妥昔单抗内吞作用的具体机制及其对西妥昔单抗摄取、生物分布和疗效的影响仍不清楚。最近，据报道他汀类药物与 EGFR 靶向药物具有协同作用。我们之前的工作表明，制霉菌素（一种胆固醇螯合抗真菌药物）通过网格蛋白依赖性途径促进内吞作用。本研究旨在调查制霉菌素是否调节西妥昔单抗和基于西妥昔单抗的抗体药物偶联物（西妥昔单抗-ADC）的摄取和功效。分别在多种人类癌细胞系和异种移植模型中研究了制霉菌素对西妥昔单抗/西妥昔单抗-ADC 的摄取和活性的体外和体内功效。我们发现，制霉菌素对胆固醇的螯合通过调节 EGFR 运输/周转并促进从脂筏向网格蛋白介导的内吞作用的转变，增强了 EGFR 阳性癌细胞中西妥昔单抗的内化。西妥昔单抗和制霉菌素联合治疗选择性增加肿瘤组织对西妥昔单抗的摄取，转化为西妥昔单抗体内增强的抗肿瘤功效（A431 和 A549 肿瘤）。制霉菌素增强的西妥昔单抗内化进一步提高了西妥昔单抗-多柔比星和西妥昔单抗-甲氨蝶呤缀合物在 EGFR 阳性西妥昔单抗耐药肿瘤中的摄取和效力。与单药西妥昔单抗或西妥昔单抗-ADC 相比，制霉菌素加西妥昔单抗或西妥昔单抗-ADC 的联合治疗进一步延长了动物存活期并显着抑制肿瘤生长。总之，我们的结果确定了一种新机制，通过胆固醇隔离增强 EGFR 靶向 mAb 和 ADC 的摄取，因此提供了临床前概念证明，即与制霉菌素联合可以增强这些 EGFR 靶向药物的递送和功效。
Cetuximab, a monoclonal antibody (mAb) targeting the epidermal growth factor receptor (EGFR), has been intensively investigated as a promising cancer treatment strategy. The specific mechanism of cetuximab endocytosis and its influence on cetuximab uptake, biodistribution and efficacy still remain elusive. Recently, statins have been reported to synergize with EGFR-targeting agents. Our prior work established that nystatin, a cholesterol-sequestering antifungal drug, facilitates endocytosis via the clathrin-dependent pathway. This study aimed to investigate whether nystatin regulates the uptake and efficacy of cetuximab and cetuximab-based antibody-drug conjugates (cetuximab-ADCs). In vitro and in vivo efficacies of nystatin on the uptake and activity of cetuximab/cetuximab-ADCs were studied in multiple human carcinoma cell lines and xenograft models, respectively. We identified that cholesterol sequestration by nystatin enhanced cetuximab internalization in EGFR-positive carcinoma cells by regulating EGFR trafficking/turnover and facilitating a switch from lipid rafts to clathrin-mediated endocytosis. Combination treatment with cetuximab and nystatin selectively increased cetuximab uptake by tumor tissues, translating into potentiated antitumor efficacy of cetuximab in vivo (A431 and A549 tumors). Nystatin-enhanced internalization of cetuximab further improved the uptake and potency of cetuximab-doxorubicin and cetuximab-methotrexate conjugates in EGFRpositive cetuximab-resistant tumors. Combination therapy with nystatin plus either cetuximab or cetuximab-ADC further prolonged animal survival and significantly suppressed tumor growth, as compared with single-agent cetuximab or cetuximab-ADC. In summary, our results identify a novel mechanism whereby cholesterol sequestration enhances the uptake of EGFR-targeting mAb and ADCs, therefore providing preclinical proof-of-concept that combination with nystatin can potentiate the delivery and efficacy of these EGFR-targeted agents.