PURPOSE Evaluating drug responses using primary patient-derived cells ex vivo represents a potentially rapid and efficient approach to screening for new treatment approaches. Here, we sought to identify neratinib combinations in HER2 mutant non-small cell lung cancer (NSCLC) patient xenograft-derived organotypic spheroids (XDOTS) using a short-term ex vivo system. EXPERIMENTAL DESIGN We generated two HER2-mutant NSCLC PDX models [DFCI359 (HER2 exon19 755_757LREdelinsRP) and DFCI315 (HER2 exon20 V777_G778insGSP)] and used the PDX tumors to generate XDOTS. Tumor spheroids were grown in a microfluidic device and treated ex vivo with neratinib-based drug combinations. Live/dead quantification was performed by dual-labeling deconvolution fluorescence microscopy. The most efficacious ex vivo combination was subsequently validated in vivo using the DFCI359 and DFCI315 PDXs and a HER2 YVMA genetically engineered mouse model. RESULTS Both neratinib and afatinib, but not gefitinib, induced cell death in DFCI359 XDOTS. The combinations of neratinib/trastuzumab and neratinib/temsirolimus enhanced the therapeutic benefit of neratinib alone in DFCI315 and DFCI359. The combination of neratinib and trastuzumab in vivo was more effective compared with single-agent neratinib or trastuzumab and was associated with more robust inhibition of HER2 and downstream signaling. CONCLUSIONS The XDOTS platform can be used to evaluate therapies and therapeutic combinations ex vivo using PDX tumors. This approach may accelerate the identification and clinical development of therapies for targets with no or few existing models and/or therapies.

中文翻译：

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使用离体患者衍生的肿瘤器官型球体来确定 HER2 突变非小细胞肺癌的联合疗法。

目的 使用离体患者原代细胞评估药物反应代表了一种潜在的快速有效的方法来筛选新的治疗方法。在这里，我们试图使用短期离体系统在 HER2 突变非小细胞肺癌 (NSCLC) 患者异种移植物衍生的器官球体 (XDOTS) 中鉴定来那替尼组合。实验设计 我们生成了两个 HER2 突变 NSCLC PDX 模型 [DFCI359 (HER2 exon19 755_757LREDelinsRP) 和 DFCI315 (HER2 exon20 V777_G778insGSP)] 并使用 PDX 肿瘤生成 XDOTS。肿瘤球体在微流体装置中生长，并用基于来那替尼的药物组合进行离体治疗。通过双标记反卷积荧光显微镜进行活/死量化。随后使用 DFCI359 和 DFCI315 PDX 以及 HER2 YVMA 基因工程小鼠模型在体内验证了最有效的离体组合。结果 在 DFCI359 XDOTS 中，来那替尼和阿法替尼均诱导细胞死亡，但吉非替尼没有。来那替尼/曲妥珠单抗和来那替尼/替西罗莫司的组合增强了来那替尼单独在 DFCI315 和 DFCI359 中的治疗益处。与单药来那替尼或曲妥珠单抗相比，来那替尼和曲妥珠单抗在体内的组合更有效，并且与更强大的 HER2 和下游信号抑制有关。结论 XDOTS 平台可用于评估使用 PDX 肿瘤的离体疗法和治疗组合。