Koch Claudia, Joosse Simon A, Schneegans Svenja, Wilken Okka J W, Janning Melanie, Loreth Desiree, Müller Volkmar, Prieske Katharina, Banys-Paluchowski Malgorzata, Horst Ludwig J, Loges Sonja, Peine Sven, Wikman Harriet, Gorges Tobias M, Pantel Klaus
Department of Tumor Biology, Center of Experimental Medicine, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany.
Department of Oncology, Hematology and Bone Marrow Transplantation with section Pneumology, Hubertus Wald Tumorzentrum, University Comprehensive Cancer Center Hamburg, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany.
Cancers (Basel). 2020 Feb 13;12(2):442. doi: 10.3390/cancers12020442.
Circulating tumor cells (CTCs) are promising tools for risk prediction and the monitoring of response to therapy in cancer patients. Within the EU/IMI CANCER-ID consortium, we validated CTC enrichment systems for future inclusion into clinical trials. Due to the known heterogeneity of markers expressed on CTCs, we tested the Parsortix system (ANGLE plc) which enables label-independent CTC enrichment from whole blood based on increased size and deformability of these tumor cells compared to leukocytes. We performed extensive comparisons both with spiked-in blood models (i.e., MDA-MB-468 tumor cell line cells spiked at very low concentration into blood from healthy donors) and validated the protocol on actual clinical samples from breast, lung, and gastrointestinal cancer patients to define optimal conditions for CTC enrichment. Multiple parameters including cassette gap, separation pressure, and cell fixatives were compared in parallel. Also, the compatibility of blood collection tubes with whole genome amplification of isolated tumor cells was demonstrated and we furthermore established a workflow for semi-automated CTC detection using a quantitative cell imager. The established workflow will contribute to supporting the use of size-based CTC enrichment platforms in clinical trials testing the clinical validity and utility of CTCs for personalized medicine.
循环肿瘤细胞（CTCs）是癌症患者风险预测和治疗反应监测的有前景的工具。在欧盟/工业医药联合会癌症识别联盟内，我们对CTC富集系统进行了验证，以便未来纳入临床试验。由于已知CTCs上表达的标志物具有异质性，我们测试了Parsortix系统（ANGLE plc公司），该系统能够基于与白细胞相比这些肿瘤细胞增大的尺寸和可变形性，从全血中进行不依赖标记的CTC富集。我们对掺入血液模型（即MDA-MB-468肿瘤细胞系细胞以非常低的浓度掺入健康供体的血液中）进行了广泛比较，并在来自乳腺癌、肺癌和胃肠道癌患者的实际临床样本上验证了方案，以确定CTC富集的最佳条件。同时比较了包括盒间隙、分离压力和细胞固定剂在内的多个参数。此外，还证明了采血管与分离肿瘤细胞的全基因组扩增的兼容性，并且我们还建立了使用定量细胞成像仪进行半自动CTC检测的工作流程。所建立的工作流程将有助于支持在测试CTCs用于个性化医疗的临床有效性和实用性的临床试验中使用基于尺寸的CTC富集平台。
