Wolfson Childhood Cancer Research Centre, Northern Institute for Cancer Research, Newcastle University, Newcastle upon Tyne, United Kingdom.
Great North Children's Hospital, Newcastle, Newcastle upon Tyne, United Kingdom.
Clin Cancer Res. 2020 Jan 1;26(1):122-134. doi: 10.1158/1078-0432.CCR-19-0656. Epub 2019 Nov 25.
Circulating tumor cells (CTCs) serve as noninvasive tumor biomarkers in many types of cancer. Our aim was to detect CTCs from patients with neuroblastoma for use as predictive and pharmacodynamic biomarkers.
We collected matched blood and bone marrow samples from 40 patients with neuroblastoma to detect GD /CD45 neuroblastoma CTCs from blood and disseminated tumor cells (DTCs) from bone marrow using the Imagestream Imaging flow cytometer (ISx). In six cases, circulating free DNA (cfDNA) extracted from plasma isolated from the CTC sample was analyzed by high-density single-nucleotide polymorphism (SNP) arrays.
CTCs were detected in 26 of 42 blood samples (1-264/mL) and DTCs in 25 of 35 bone marrow samples (57-291,544/mL). Higher numbers of CTCs in patients with newly diagnosed, high-risk neuroblastoma correlated with failure to obtain a complete bone marrow (BM) metastatic response after induction chemotherapy ( < 0.01). Nutlin-3 (MDM2 inhibitor) treatment of blood and BM increased p53 and p21 expression in CTCs and DTCs compared with DMSO controls. In five of six cases, cfDNA analyzed by SNP arrays revealed copy number abnormalities associated with neuroblastoma.
This is the first study to show that CTCs and DTCs are detectable in neuroblastoma using the ISx, with concurrently extracted cfDNA used for copy number profiling, and may be useful as pharmacodynamic biomarkers in early-phase clinical trials. Further investigation is required to determine whether CTC numbers are predictive biomarkers of BM response to first-line induction chemotherapy.
循环肿瘤细胞（CTC）可作为多种癌症的非侵入性肿瘤生物标志物。我们的目的是从神经母细胞瘤患者中检测 CTCs，将其用作预测和药效动力学生物标志物。
我们收集了 40 名神经母细胞瘤患者的配对血液和骨髓样本，使用 ISx （ Imagestream 成像流式细胞仪）从血液中检测 GD/CD45 神经母细胞瘤 CTCs ，从骨髓中检测播散性肿瘤细胞（DTCs）。在 6 例患者中，从 CTC 样本分离的血浆中提取的循环游离 DNA（cfDNA）通过高密度单核苷酸多态性（SNP）阵列进行分析。
在 42 份血液样本（1-264/mL）中有 26 份检测到 CTCs ，在 35 份骨髓样本（57-291,544/mL）中有 25 份检测到 DTCs。初诊时高风险神经母细胞瘤患者的 CTC 数量较多，与诱导化疗后未能获得完全骨髓（BM）转移反应相关（<0.01）。与 DMSO 对照相比，MDM2 抑制剂（Nutlin-3）对血液和 BM 的处理增加了 CTCs 和 DTCs 中 p53 和 p21 的表达。在 6 例患者中，通过 SNP 阵列分析的 cfDNA 揭示了与神经母细胞瘤相关的拷贝数异常。
这是第一项使用 ISx 检测神经母细胞瘤中 CTCs 和 DTCs 的研究，同时提取的 cfDNA 用于拷贝数分析，可能作为早期临床试验中的药效动力学生物标志物有用。需要进一步研究来确定 CTC 数量是否是对一线诱导化疗 BM 反应的预测性生物标志物。
