黑色素瘤治疗方法的进步迫切需要生物标志物来识别有复发风险的患者并指导治疗。液体活检在识别生物标志物方面的潜在用途越来越受到人们的认可。在这里，我们对几种分析 2​​0 名转移性黑色素瘤患者的循环肿瘤细胞 (CTC) 和游离 DNA (cfDNA) 的技术进行了头对头比较。在本研究中，我们研究了与 CellSearch 平台相比，诊断性白细胞分离术 (DLA) 与多标记流式细胞术 (FCM) 相结合是否可以增加血液中 CTC 的检测。此外，我们在体细胞突变、非整倍性程度和全基因组 DNA 甲基化水平上对 cfDNA 进行了表征。将 CTC 和 cfDNA 测量值与放射成像的肿瘤标志物和颅外肿瘤负荷进行比较。与应用于外周血的CellSearch方法相比，DLA联合FCM将可检测到CTC的患者比例从35%提高到70%（P = 0.06）。然而，通过 DLA 程序可以回收的细胞百分比中位数为 29%。或者，cfDNA 突变和甲基化分析检测到大多数患者的肿瘤负荷（分别成功分析了 90% 和 93% 的样本）。 35% 的患者非整倍体评分呈阳性。从血液中的所有肿瘤测量结果来看，乳酸脱氢酶 (LDH) 水平与变异等位基因频率显着相关 (P = 0.004)。此外，DLA 中 CTC 的存在与肿瘤负荷相关（P<0.001），而外周血中 CTC 的存在与放射成像病变数量相关（P<0.001）。总之，DLA 往往会增加可检测到 CTC 的患者比例，但也与康复率较低有关。 cfDNA 和 CTC 均与 LDH 水平和颅外肿瘤负荷等临床参数相关。© 2024 作者。约翰·威利出版的《分子肿瘤学》

Advances in therapeutic approaches for melanoma urge the need for biomarkers that can identify patients at risk for recurrence and to guide treatment. The potential use of liquid biopsies in identifying biomarkers is increasingly being recognized. Here, we present a head-to-head comparison of several techniques to analyze circulating tumor cells (CTCs) and cell-free DNA (cfDNA) in 20 patients with metastatic melanoma. In this study, we investigated whether diagnostic leukapheresis (DLA) combined with multimarker flow cytometry (FCM) increased the detection of CTCs in blood compared to the CellSearch platform. Additionally, we characterized cfDNA at the level of somatic mutations, extent of aneuploidy and genome-wide DNA methylation. Both CTCs and cfDNA measures were compared to tumor markers and extracranial tumor burden on radiological imaging. Compared to the CellSearch method applied on peripheral blood, DLA combined with FCM increased the proportion of patients with detectable CTCs from 35% to 70% (P = 0.06). However, the median percentage of cells that could be recovered by the DLA procedure was 29%. Alternatively, cfDNA mutation and methylation analysis detected tumor load in the majority of patients (90% and 93% of samples successfully analyzed, respectively). The aneuploidy score was positive in 35% of all patients. From all tumor measurements in blood, lactate dehydrogenase (LDH) levels were significantly correlated to variant allele frequency (P = 0.004). Furthermore, the presence of CTCs in DLA was associated with tumor burden (P < 0.001), whereas the presence of CTCs in peripheral blood was associated with number of lesions on radiological imaging (P < 0.001). In conclusion, DLA tended to increase the proportion of patients with detectable CTCs but was also associated with low recovery. Both cfDNA and CTCs were correlated with clinical parameters such as LDH levels and extracranial tumor burden.© 2024 The Authors. Molecular Oncology published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.