Abstract：
Purpose: Antibody-drug conjugates (ADCs) are a relatively recent advance in the delivery of chemotherapeutics that improve targeting of cytotoxic agents. However, despite their antitumor activity, severe ocular adverse effects, including vision loss, have been reported for several ADCs. The nonspecific uptake of ADCs into human corneal epithelial cells (HCECs) and their precursors via macropinocytosis has been proposed to be the primary mechanism of ocular toxicity. In this study, we evaluated the ability of a novel polymer, poly(l-lysine)-graft-poly(ethylene glycol) (PLL-g-PEG), to decrease the ADC rituximab-mc monomethylauristatin F (MMAF) (RIX) uptake into human corneal epithelial (HCE-T) cells. Methods: HCE-T cells were exposed to increasing concentrations of RIX to determine inhibition of cell proliferation. HCE-T cells were treated with PLL-g-PEG, the macropinocytosis inhibitor 5-(N-ethyl-N-isopropyl) amiloride (EIPA), or vehicle. After 30 min of incubation, RIX was added. ADC was detected by fluorescent anti-human immunoglobulin G and fluorescently conjugated dextran as viewed by microscopy. Results: RIX caused dose-dependent inhibition of HCE-T cell proliferation. EIPA significantly reduced RIX uptake and decreased macropinocytosis as assessed by direct quantification of RIX using a fluorescently conjugated anti-human antibody as well as quantification of macropinocytosis using fluorescently conjugated dextran. PLL-g-PEG resulted in a dose-dependent inhibition of RIX uptake with half-maximal inhibitory concentrations of 0.022%-0.023% PLL-g-PEG. Conclusion: The data show PLL-g-PEG to be a potent inhibitor of RIX uptake by corneal epithelial cells and support its use as a novel therapeutic approach for the prevention of ocular adverse events associated with ADC therapy.
摘要：
目的：抗体-药物缀合物（ADC）是化疗药物递送方面相对较新的进展，可改善细胞毒性剂的靶向性。然而,尽管它们具有抗肿瘤活性，严重的眼部不良反应，包括视力丧失,已经报道了几个ADC。已提出通过大细胞胞吞作用将ADC非特异性摄取到人角膜上皮细胞（HCECs）及其前体中是眼部毒性的主要机制。在这项研究中,我们评估了一种新型聚合物的能力，聚(l-赖氨酸)-接枝-聚(乙二醇)(PLL-g-PEG)，降低ADC利妥昔单抗-mc单甲基立他汀F（MMAF）（RIX）对人角膜上皮（HCE-T）细胞的摄取。方法:将HCE-T细胞暴露于增加浓度的RIX以确定细胞增殖的抑制作用。HCE-T细胞用PLL-g-PEG处理,巨噬细胞吞噬抑制剂5-(N-乙基-N-异丙基)阿米洛利(EIPA)，或车辆。孵育30分钟后，添加了RIX。如通过显微镜观察,通过荧光抗人免疫球蛋白G和荧光缀合的葡聚糖检测ADC。结果:RIX引起HCE-T细胞增殖的剂量依赖性抑制。如通过使用荧光缀合的抗人抗体对RIX的直接定量以及使用荧光缀合的葡聚糖对巨成细胞作用的定量所评估的,EIPA显著降低RIX摄取和降低巨成细胞作用。PLL-g-PEG导致RIX摄取的剂量依赖性抑制，其最大抑制浓度为0.022％-0.023％PLL-g-PEG。结论：数据显示PLL-g-PEG是角膜上皮细胞摄取RIX的有效抑制剂，并支持将其用作预防与ADC治疗相关的眼部不良事件的新型治疗方法。
