Wang Chunming, Luo Qiong, Huang Wenbin, Zhang Cheng, Liao Hangyu, Chen Kunling, Pan MingXin
General Surgery Center, Department of Hepatobiliary Surgery II, Guangdong Provincial Research Center for Artificial Organ and Tissue Engineering, Guangzhou Clinical Research and Transformation Center for Artificial Liver, Institute of Regenerative Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou, China.
Department of General Surgery, Affiliated Hengyang Hospital, Southern Medical University (Hengyang Central Hospital), Hengyang, China.
Front Oncol. 2021 Jun 11;11:686365. doi: 10.3389/fonc.2021.686365. eCollection 2021.
Liquid biopsy is attracting attention as a method of real-time monitoring of patients with tumors. It can be used to understand the temporal and spatial heterogeneity of tumors and has good clinical application prospects. We explored a new type of circulating tumor cell (CTC) enrichment technology combined with next-generation sequencing (NGS) to analyze the correlation between genomic alterations in circulating tumor cells of hepatocellular carcinoma and the counts of mesenchymal CTCs and CTC-associated white blood cell (CTC-WBC) clusters.
We collected peripheral blood samples from 29 patients with hepatocellular carcinoma from January 2016 to December 2019. We then used the CanPatrol™ system to capture and analyze mesenchymal CTCs and CTC-WBC clusters for all the patients. A customized Illumina panel was used for DNA sequencing and the Mann-Whitney U test was used to test the correlation between mesenchymal CTCs, CTC-WBC cluster counts, and specific genomic changes.
At least one somatic hotspot mutation was detected in each of the 29 sequenced patients. A total of 42 somatic hot spot mutations were detected in tumor tissue DNA, and 39 mutations were detected in CTC-DNA, all of which included common changes in PTEN, MET, EGFR, RET, and FGFR3. The number of mesenchymal CTCs was positively correlated with the somatic genomic alterations in the PTEN and MET genes (PTEN, P = 0.021; MET, P = 0.008, Mann-Whitney U test) and negatively correlated with the somatic genomic alterations in the EGFR gene (P = 0.006, Mann-Whitney U test). The number of CTC-WBC clusters was positively correlated with the somatic genomic alterations in RET genes (P = 0.01, Mann-Whitney U test) and negatively correlated with the somatic genomic alterations in FGFR3 (P = 0.039, Mann-Whitney U test).
We report a novel method of a CTC enrichment platform combined with NGS technology to analyze genetic variation, which further demonstrates the potential clinical application of this method for spatiotemporal heterogeneity monitoring of hepatocellular carcinoma. We found that the number of peripheral blood mesenchymal CTCs and CTC-WBC clusters in patients with hepatocellular carcinoma was related to a specific genome profile.
液体活检作为一种对肿瘤患者进行实时监测的方法正受到关注。它可用于了解肿瘤的时空异质性，具有良好的临床应用前景。我们探索了一种新型的循环肿瘤细胞（CTC）富集技术，并结合下一代测序（NGS）来分析肝细胞癌循环肿瘤细胞中的基因组改变与间充质CTC及CTC相关白细胞（CTC-WBC）簇计数之间的相关性。
我们收集了2016年1月至2019年12月期间29例肝细胞癌患者的外周血样本。然后使用CanPatrol™系统对所有患者的间充质CTC和CTC-WBC簇进行捕获和分析。使用定制的Illumina面板进行DNA测序，并使用曼-惠特尼U检验来检测间充质CTC、CTC-WBC簇计数与特定基因组变化之间的相关性。
在29例测序患者中，每例均检测到至少一个体细胞热点突变。在肿瘤组织DNA中共检测到42个体细胞热点突变，在CTC-DNA中检测到39个突变，所有这些突变均包括PTEN、MET、EGFR、RET和FGFR3的常见变化。间充质CTC的数量与PTEN和MET基因中的体细胞基因组改变呈正相关（PTEN，P = 0.021；MET，P = 0.008，曼-惠特尼U检验），与EGFR基因中的体细胞基因组改变呈负相关（P = 0.006，曼-惠特尼U检验）。CTC-WBC簇的数量与RET基因中的体细胞基因组改变呈正相关（P = 0.01，曼-惠特尼U检验），与FGFR3中的体细胞基因组改变呈负相关（P = 0.039，曼-惠特尼U检验）。
我们报告了一种结合NGS技术的CTC富集平台分析基因变异的新方法，这进一步证明了该方法在肝细胞癌时空异质性监测中的潜在临床应用价值。我们发现肝细胞癌患者外周血间充质CTC和CTC-WBC簇的数量与特定的基因组图谱相关。
