Clinical evaluation of a novel microfluidic device for epitope-independent enrichment of circulating tumour cells in patients with small cell lung cancer
Clinical evaluation of a novel microfluidic device for epitope-independent enrichment of circulating tumour cells in patients with small cell lung cancer
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DOI：
10.1039/c5an02156a
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发表时间：
2016-01-01
期刊：
影响因子：
4.2
通讯作者：
Brady, Ged
中科院分区：
文献类型：
--
作者：
Chudziak, Jakub;Burt, Deborah J.;Brady, Ged
循环肿瘤细胞（CTC）具有作为微创生物标志物的潜在效用，以帮助癌症治疗决策。然而，许多目前的CTC技术使用不一定反映CTC异质性的特异性表面表位来富集CTC。在这里，我们评估了表位独立的Parsortix系统，该系统使用掺入肿瘤细胞的健康正常志愿者血液样品和来自小细胞肺癌（SCLC）患者的血液样品基于大小和刚性富集CTC。将血液样品在室温下保持不分级最多4天，然后除去血浆以分离循环游离DNA（cfDNA），并将剩余的细胞组分直接应用于Parsortix系统。对于表达EpCAM细胞表面标志物的肿瘤细胞，使用Parsortix系统和通过使用Cell-Search（R）的EpCAM阳性选择保留的掺入细胞的数量没有显著差异，而只有Parsortix系统显示出具有不可检测的EpCAM表达的细胞的强烈富集。在初步临床研究中，我们储存了富集的CTC以及来自SCLC患者血液样品的血浆。在检索库存的Parsortix细胞样品后，我们可以在所有12名测试的SCLC患者中检测到细胞角蛋白阳性CTC。有趣的是，使用CellSearch（R）通过EpCAM富集处理来自相同患者的平行样品显示仅83%（10/12）具有细胞角蛋白阳性CTC，表明Parsortix系统富集EpCAM阴性SCLC CTC。我们的组合结果表明Parsortix系统是用于组合cfDNA分离和CTC富集的有价值的工具，其使得CTC分析能够扩展到对表面表位的依赖之外。
Circulating tumour cells (CTCs) have potential utility as minimally-invasive biomarkers to aid cancer treatment decision making. However, many current CTC technologies enrich CTCs using specific surface epitopes that do not necessarily reflect CTC heterogeneity. Here we evaluated the epitope-independent Parsortix system which enriches CTCs based on size and rigidity using both healthy normal volunteer blood samples spiked with tumour cells and blood samples from patients with small cell lung cancer (SCLC). Blood samples were maintained unfractionated at room temperature for up to 4 days followed by plasma removal for circulating free DNA (cfDNA) isolation and direct application of the remaining cell component to the Parsortix system. For tumour cells expressing the EpCAM cell surface marker the numbers of spiked cells retained using the Parsortix system and by EpCAM-positive selection using Cell-Search (R) were not significantly different, whereas only the Parsortix system showed strong enrichment of cells with undetectable EpCAM expression. In a pilot clinical study we banked both enriched CTCs as well as plasma from SCLC patient blood samples. Upon retrieval of the banked Parsortix cellular samples we could detect cytokeratin positive CTCs in all 12 SCLC patients tested. Interestingly, processing parallel samples from the same patients by EpCAM enrichment using CellSearch (R) revealed only 83% (10/12) with cytokeratin positive CTCs indicating the Parsortix system is enriching for EpCAM negative SCLC CTCs. Our combined results indicate the Parsortix system is a valuable tool for combined cfDNA isolation and CTC enrichment that enables CTC analysis to be extended beyond dependence on surface epitopes.