背景:由非小细胞肺癌(NSCLC)引起的复发性恶性胸腔积液(MPE)对常规疗法很难治疗,并且缺乏预测性标志物。复发性MPE的细胞或遗传特征仍然很大程度上不确定。
方法:纳入16例有胸腔积液的非小细胞肺癌患者,然后根据原发性肿瘤进行相应的治疗。治疗3-6周后确定非复发性或复发性MPE。通过计算机断层扫描(CT)和细胞病理学检查证实了MPE的状态,收集基线胸水进行单细胞RNA测序(scRNA-seq).然后对样品进行整合和轮廓分析。通过生物信息学算法推断细胞通信和轨迹。进行了比较分析,并通过定量聚合酶链反应(qPCR)在作者中心(n=64)的较大MPE队列中进一步验证了结果。
结果:scRNA-seq显示33.590细胞被注释为7种主要细胞类型,并进一步精确表征为14个细胞簇。细胞簇C1,分类为上皮细胞粘附分子(EpCAM)转移癌细胞,与紧密连接和粘附连接的激活相关,在复发性MPE组中显著富集,其中确定了Claudin-4(CLDN4)。C1的亚群细胞簇C3在复发性MPE中富集,并表现出异形型细胞迁移的表型,也显示了CLDN4的显著较高的表达。同时,CLDN4的表达与E74样ETS转录因子3(ELF3)呈正相关,EpCAM和肿瘤相关钙信号传感器2(TACSTD2),独立于驱动基因状态。还发现CLDN4与缺氧诱导因子1亚基α(HIF1A)和血管内皮生长因子A(VEGFA)的表达有关,细胞簇C1是VEGFA信号的细胞通讯的主要介质。在广泛的MPE队列中,在诊断为复发性MPE的患者中,观察到胸腔积液细胞中CLDN4的表达显着增加,与非复发组相比,这也与总体生存率(OS)下降的趋势有关。
结论:CLDN4可作为晚期NSCLC患者MPE复发的预测指标。有必要在样本量较大的队列中进一步验证其临床价值,并对其生物学功能进行深入的机制研究。
背景:不适用。
Recurrent malignant pleural effusion (MPE) resulting from non-small-cell lung cancer (NSCLC) is easily refractory to conventional therapeutics and lacks predictive markers. The cellular or genetic signatures of recurrent MPE still remain largely uncertain.
16 NSCLC patients with pleural effusions were recruited, followed by corresponding treatments based on primary tumours. Non-recurrent or recurrent MPE was determined after 3-6 weeks of treatments. The status of MPE was verified by computer tomography (CT) and cytopathology, and the baseline pleural fluids were collected for single-cell RNA sequencing (scRNA-seq). Samples were then integrated and profiled. Cellular communications and trajectories were inferred by bioinformatic algorithms. Comparative analysis was conducted and the results were further validated by quantitative polymerase chain reaction (qPCR) in a larger MPE cohort from the authors\' centre (n = 64).
The scRNA-seq revealed that 33 590 cells were annotated as 7 major cell types and further characterized into 14 cell clusters precisely. The cell cluster C1, classified as Epithelial Cell Adhesion Molecule (EpCAM)+ metastatic cancer cell and correlated with activation of tight junction and adherence junction, was significantly enriched in the recurrent MPE group, in which Claudin-4 (CLDN4) was identified. The subset cell cluster C3 of C1, which was enriched in recurrent MPE and demonstrated a phenotype of ameboidal-type cell migration, also showed a markedly higher expression of CLDN4. Meanwhile, the expression of CLDN4 was positively correlated with E74 Like ETS Transcription Factor 3 (ELF3), EpCAM and Tumour Associated Calcium Signal Transducer 2 (TACSTD2), independent of driver-gene status. CLDN4 was also found to be associated with the expression of Hypoxia Inducible Factor 1 Subunit Alpha (HIF1A) and Vascular Endothelial Growth Factor A (VEGFA), and the cell cluster C1 was the major mediator in cellular communication of VEGFA signalling. In the extensive MPE cohort, a notably increased expression of CLDN4 in cells from pleural effusion among patients diagnosed with recurrent MPE was observed, compared with the non-recurrent group, which was also associated with a trend towards worse overall survival (OS).
CLDN4 could be considered as a predictive marker of recurrent MPE among patients with advanced NSCLC. Further validation for its clinical value in cohorts with larger sample size and in-depth mechanism studies on its biological function are warranted.
Not applicable.