Ex Vivo Expanded Circulating Tumor Cells for Clinical Anti-Cancer Drug Prediction in Patients with Head and Neck Cancer

体外扩增循环肿瘤细胞用于头颈癌患者的临床抗癌药物预测

The conventional methods that seek to predict clinical treatment response are based on the number of circulating tumor cells (CTCs) present in liquid biopsies or genetic profiling of extracted CTCs. This paper presents a novel process by which CTCs can be extracted from blood samples taken from head and neck cancer patients and then expanded ex vivo to form organoids that can be tested with a panel of anti-cancer treatments. The resulting drug sensitivity profiles derived from cisplatin treatment of organoids were subsequently found to correlate with clinical treatment response to cisplatin in patients. CTCs extracted from liquid biopsies for ex vivo expansion negates the need for complicated and potentially risky biopsies of tumor material, thereby supporting the application of this procedure for checkups and treatment monitoring.

寻求预测临床治疗反应的传统方法是基于液体活检中存在的循环肿瘤细胞 (CTC) 的数量或提取的 CTC 的基因图谱。本文介绍了一种新的方法，通过该方法可以从头颈癌患者的血液样本中提取 CTC，然后在体外扩增形成可以用一组抗癌治疗进行测试的类器官。随后发现，源自类器官顺铂治疗的药物敏感性曲线与患者对顺铂的临床治疗反应相关。从液体活检中提取的用于离体扩增的 CTC 无需对肿瘤材料进行复杂且具有潜在风险的活检，从而支持该程序在检查和治疗监测中的应用。

三角细胞摇瓶

The advanced-stage head and neck cancer (HNC) patients respond poorly to platinum-based treatments. Thus, a reliable pretreatment method for evaluating platinum treatment response would improve therapeutic efficiency and outcomes. This study describes a novel strategy to predict clinical drug responses in HNC patients by using eSelect, a lab-developed biomimetic cell culture system, which enables us to perform ex vivo expansion and drug sensitivity profiling of circulating tumor cells (CTCs). Forty liquid biopsies were collected from HNC patients, and the CTCs were expanded ex vivo using the eSelect system within four weeks. Immunofluorescence staining confirmed that the CTC-derived organoids were positive for EpCAM and negative for CD45. Two illustrative cases present the potential of this strategy for evaluating treatment response. The statistical analysis confirmed that drug sensitivity in CTC-derived organoids was associated with a clinical response. The multivariant logistic regression model predicted that the treatment accuracy of chemotherapy responses achieved 93.75%, and the area under the curves (AUCs) of prediction models was 0.8841 in the whole dataset and 0.9167 in cisplatin specific dataset. In summary, cisplatin sensitivity profiles of patient-derived CTCs expanded ex vivo correlate with a clinical response to cisplatin treatment, and this can potentially underpin predictive assays to guide HNC treatments.

血清培养基瓶

晚期头颈癌 (HNC) 患者对铂类治疗反应不佳。因此，用于评估铂类治疗反应的可靠预处理方法将提高治疗效率和结果。本研究描述了一种通过使用 eSelect 预测 HNC 患者临床药物反应的新策略，eSelect 是一种实验室开发的仿生细胞培养系统，使我们能够对循环肿瘤细胞 (CTC) 进行离体扩增和药物敏感性分析。从 HNC 患者中收集了 40 份液体活检，并在 4 周内使用 eSelect 系统离体扩增了 CTC。免疫荧光染色证实 CTC 衍生的类器官对 EpCAM 呈阳性，对 CD45 呈阴性。两个说明性案例展示了这种策略评估治疗反应的潜力。统计分析证实，CTC 衍生的类器官中的药物敏感性与临床反应相关。多变量逻辑回归模型预测化疗反应的治疗准确率达到93.75%，预测模型的曲线下面积（AUCs）在整个数据集中为0.8841，在顺铂特异性数据集中为0.9167。总之，患者衍生的 CTC 的顺铂敏感性分布与体外扩展的顺铂治疗的临床反应相关，这可能支持预测性分析以指导 HNC 治疗。预测模型的曲线下面积（AUCs）在整个数据集中为 0.8841，在顺铂特定数据集中为 0.9167。总之，患者衍生的 CTC 的顺铂敏感性分布与体外扩展的顺铂治疗的临床反应相关，这可能支持预测性分析以指导 HNC 治疗。预测模型的曲线下面积（AUCs）在整个数据集中为 0.8841，在顺铂特定数据集中为 0.9167。总之，患者衍生的 CTC 的顺铂敏感性分布与体外扩展的顺铂治疗的临床反应相关，这可能支持预测性分析以指导 HNC 治疗。

细胞工厂

In conclusion, this study developed an efficient and reliable method by which CTCs from liquid biopsies of patients with locally advanced HNC patients can be isolated and expanded ex vivo using the eSelect system. Drug sensitivity assays conducted on the expanded CTC-derived organoids were subsequently correlated with clinical response in patients with HNC. The results highlight the potential of this method to assist clinicians in assessing and predicting treatment response in HNC, and more extensive prospective controlled studies are warranted to validate these findings further and to improve this process for future application in clinical settings.

总之，本研究开发了一种有效且可靠的方法，通过该方法，可以使用 eSelect 系统从局部晚期 HNC 患者的液体活检中分离和扩增 CTC。随后对扩展的 CTC 衍生类器官进行的药物敏感性测定与 HNC 患者的临床反应相关。结果突出了这种方法在帮助临床医生评估和预测 HNC 治疗反应方面的潜力，需要进行更广泛的前瞻性对照研究来进一步验证这些发现并改进这一过程，以便将来在临床环境中应用。

关键词： circulating tumor cells,drug sensitivity,ex vivo expansion,head and neck cancer,response to therapy,循环肿瘤细胞,药物敏感性,离体扩增,头颈癌,对治疗的反应

来源：MDPI https://www.mdpi.com/2072-6694/13/23/6076/htm