Markou A N, Londra D, Stergiopoulou D, Vamvakaris I, Potaris K, Pateras I S, Kotsakis A, Georgoulias V, Lianidou E
Lab of Analytical Chemistry, Department of Chemistry, National and Kapodistrian University of Athens, 15771 Athens, Greece.
Department of Pathology; 'Sotiria' General Hospital for Chest Diseases, 11527 Athens, Greece.
Cancers (Basel). 2023 Mar 21;15(6):1877. doi: 10.3390/cancers15061877.
We assessed whether preoperativemutational analyses of circulating tumor cells (CTCs) and plasma-cfDNA could be used as minimally invasive biomarkers and as complimentary tools for early prediction of relapse in early-stage non-small -cell lung cancer (NSCLC).
Using ddPCR assays, hotspot mutations of and were identified in plasma-cfDNA samples and size-based enriched CTCs isolated from the same blood samples of 49 early-stage NSCLC patients before surgery and in a control group of healthy blood donors (= 22). Direct concordance of the mutational spectrum was further evaluated in 27 patient-matched plasma-cfDNA and CTC-derived DNA in comparison to tissue-derived DNA.
The prevalence of detectable mutations of the four tested genes was higher in CTC-derived DNA than in the corresponding plasma-cfDNA (38.8% and 24.5%, respectively).The most commonly mutated gene was PIK3CA, in both CTCs and plasma-cfDNA at baseline and at the time of relapse. Direct comparison of the mutation status of selected drug-responsive genes in CTC-derived DNA, corresponding plasma-cfDNA and paired primary FFPE tissues clearly showed the impact of heterogeneity both within a sample type, as well as between different sample components. The incidence of relapse was higher when at least one mutation was detected in CTC-derived DNA or plasma-cfDNA compared with patients in whom no mutation was detected ( =0.023). Univariate analysis showed a significantly higher risk of progression (HR: 2.716; 95% CI, 1.030-7.165; =0.043) in patients with detectable mutations in plasma-cfDNA compared with patients with undetectable mutations, whereas the hazard ratio was higher when at least one mutation was detected in CTC-derived DNA or plasma-cfDNA (HR: 3.375; 95% CI, 1.098-10.375; =0.034).
Simultaneous mutational analyses of plasma-cfDNA and CTC-derived DNA provided complementary molecular information from the same blood sample and greater diversity in genomic information for cancer treatment and prognosis. The detection of specific mutations in ctDNA and CTCs in patients with early-stage NSCLC before surgery was independently associated with disease recurrence, which represents an important stratification factor for future trials.
我们评估了循环肿瘤细胞（CTC）和血浆游离DNA（cfDNA）的术前突变分析是否可作为微创生物标志物以及作为早期预测早期非小细胞肺癌（NSCLC）复发的补充工具。
使用数字滴度PCR（ddPCR）检测法，在49例早期NSCLC患者术前采集的相同血样以及22例健康献血者对照组的血浆cfDNA样本和基于大小富集的CTC中，鉴定了 和 的热点突变。与组织来源的DNA相比，进一步评估了27例患者匹配的血浆cfDNA和CTC衍生DNA中突变谱的直接一致性。
四个检测基因的可检测突变在CTC衍生DNA中的发生率高于相应的血浆cfDNA（分别为38.8%和24.5%）。最常发生突变的基因是PIK3CA，在基线期和复发时的CTC和血浆cfDNA中均如此。对CTC衍生DNA、相应血浆cfDNA和配对的原发性福尔马林固定石蜡包埋（FFPE）组织中选定的药物反应基因的突变状态进行直接比较，清楚地显示了样本类型内以及不同样本成分之间的异质性影响。与未检测到突变的患者相比，在CTC衍生DNA或血浆cfDNA中检测到至少一个突变的患者复发率更高（P = 0.023）。单因素分析显示，与未检测到突变的患者相比，血浆cfDNA中检测到可检测突变的患者进展风险显著更高（风险比：2.716；95%置信区间，1.030 - 7.165；P = 0.043），而当在CTC衍生DNA或血浆cfDNA中检测到至少一个突变时，风险比更高（风险比：3.375；95%置信区间，1.098 - 10.375；P = 0.034）。
血浆cfDNA和CTC衍生DNA的同步突变分析从同一血样中提供了互补的分子信息，并在癌症治疗和预后的基因组信息方面具有更大的多样性。术前早期NSCLC患者的循环肿瘤DNA（ctDNA）和CTC中特定突变的检测与疾病复发独立相关，这代表了未来试验的一个重要分层因素。
