PO.ET02.06 · 实验与分子治疗
对HER2靶向抗体-药物偶联物疗法进行实时、无标记评估
Real-time, label-free assessment of HER2-targeted antibody-drug conjugate therapies
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
抗体-药物偶联物（ADC）由单克隆抗体与小分子药物共价连接而成，二者结合可增强双方的抗肿瘤效果。多种ADC靶向人表皮生长因子受体2（HER2），这是一种在许多癌症中过表达的细胞表面受体。其在肿瘤细胞中的表达增加使HER2成为细胞毒性疗法极具吸引力的靶点，从而能够特异性杀伤癌细胞，同时最大限度地减少脱靶效应。在此，我们开发了一种体外试验，用于测量使用HER2靶向单克隆抗体trastuzumab的ADC的细胞毒能力。该试验采用Maestro Z平台以实时、无标记的方式进行。Maestro Z通过嵌入孔表面的电极测量电阻抗，以监测细胞生长和增殖。为评估一种HER2靶向ADC——trastuzumab-deruxtecan（DS-8201a，商品名ENHERTU®）的细胞毒效应，将5,000个HER2阳性SKOV3细胞接种于CytoView Z板中，监测24小时后给予递增剂量的DS-8201a。SKOV3细胞以剂量依赖方式被杀伤，较高剂量的DS-8201a导致SKOV3阻抗更大幅度的下降，表明SKOV3死亡更多。在最高剂量下，给药后约5天SKOV3细胞裂解率达到90.9%，计算得出EC50值为30.7 µg/mL。我们还测试了DS-8201a对其他具有不同HER2表达程度细胞类型的效应，包括A549（HER2低表达）和MDA-MB-231（无HER2表达）细胞。正如预期，阻抗数据显示，与其他两种HER2表达较低的细胞类型相比，DS-8201a对高HER2表达的SKOV3细胞具有增强的细胞毒效应。最后，我们在Maestro Z上比较了DS-8201a与另一种基于trastuzumab的ADC——trastuzumab-emtansine（T-DM1，商品名Kadcyla®）的细胞毒效应。总之，这些结果表明，可使用Maestro Z筛选ADC疗法对HER2阳性细胞类型的细胞毒能力。此处所展示的试验可用于进一步开发针对HER2阳性癌症的ADC疗法，以及针对其他癌症特异性靶点的ADC。
查看英文原文 English abstract
Antibody-drug conjugates (ADCs) are made of monoclonal antibodies covalently linked to small molecule drugs that, together, can improve both components' anti-tumor effects. Several ADCs are targeted towards human epidermal growth factor receptor 2 (HER2), a cell surface receptor that is overexpressed in many cancers. Its increased expression in tumor cells makes HER2 an attractive target for cytotoxic therapies, allowing for the specific killing of cancer cells while minimizing off-target effects. Here, we developed an in vitro assay to measure the cytotoxic capacity of ADCs that use the HER2-targeted monoclonal antibody trastuzumab. This assay was performed in a real-time, label-free manner using the Maestro Z platform. The Maestro Z measures electrical impedance via electrodes embedded in the surface of the well to monitor cell growth and proliferation. To assess the cytotoxic effects of one HER2-targeted ADC, trastuzumab-deruxtecan (DS-8201a, branded as ENHERTU®), 5,000 HER2-positive SKOV3 cells were plated in CytoView Z plates and monitored for 24 hours before being dosed with increasing amounts of DS-8201a. SKOV3 cells were killed in a dose-dependent manner, and higher doses of DS-8201a led to larger decreases in SKOV3 impedance, indicative of greater SKOV3 death. SKOV3 cytolysis at the highest dose reached 90.9% at ~ 5 days post dose, and the EC50 value was calculated to be 30.7 µg/mL. We also tested the effects of DS-8201a against other cell types with varying degrees of HER2 expression, including A549 (low HER2 expression) and MDA-MB-231 (no HER2 expression) cells. As expected, impedance data showed that DS-8201a had enhanced cytotoxic effects on the high HER2-expressing SKOV3 cells compared to the other two cell types with lower HER2 expression. Finally, we compared the cytotoxic effects of DS-8201a to another trastuzumab-based ADC, trastuzumab-emtasine (T-DM1, branded as Kadcyla®) on the Maestro Z. Taken together, these results show that the cytotoxic capacity of ADC therapies toward HER2 positive cell types can be screened using the Maestro Z. The assay presented here can be used to further develop ADC therapies aimed at HER2-positive cancers as well as ADCs toward other cancer-specific targets.
利益披露 Disclosure
D. Flanigan, None.