Institute for Molecules and Materials, Radboud University, Heyendaalseweg 135, 6525 AJ Nijmegen, the Netherlands.
Department of Medical Oncology, Radboud University Medical Center, Nijmegen, the Netherlands.
Sci Adv. 2020 Sep 30;6(40). doi: 10.1126/sciadv.aaz3849. Print 2020 Sep.
Despite their important role in metastatic disease, no general method to detect circulating stromal cells (CStCs) exists. Here, we present the Metabolic Assay-Chip (MA-Chip) as a label-free, droplet-based microfluidic approach allowing single-cell extracellular pH measurement for the detection and isolation of highly metabolically active cells (hm-cells) from the tumor microenvironment. Single-cell mRNA-sequencing analysis of the hm-cells from metastatic prostate cancer patients revealed that approximately 10% were canonical EpCAM hm-CTCs, 3% were EpCAM hm-CTCs with up-regulation of prostate-related genes, and 87% were hm-CStCs with profiles characteristic for cancer-associated fibroblasts, mesenchymal stem cells, and endothelial cells. Kaplan-Meier analysis shows that metastatic prostate cancer patients with more than five hm-cells have a significantly poorer survival probability than those with zero to five hm-cells. Thus, prevalence of hm-cells is a prognosticator of poor outcome in prostate cancer, and a potentially predictive and therapy response biomarker for agents cotargeting stromal components and preventing epithelial-to-mesenchymal transition.
尽管它们在转移性疾病中起着重要作用，但目前还没有一种通用的方法来检测循环基质细胞（CStCs）。在这里，我们提出了代谢分析芯片（MA-Chip），这是一种无标记的基于液滴的微流控方法，允许单细胞细胞外 pH 值测量，用于从肿瘤微环境中检测和分离高度代谢活跃的细胞（hm-cells）。对转移性前列腺癌患者的 hm-cells 进行单细胞 mRNA 测序分析表明，大约 10%是典型的 EpCAM hm-CTC，3%是 EpCAM hm-CTC 上调与前列腺相关的基因，87%是 hm-CStCs，具有与癌症相关的成纤维细胞、间充质干细胞和内皮细胞特征的特征。Kaplan-Meier 分析表明，转移性前列腺癌患者中 hm-cells 超过 5 个的患者的生存概率明显低于 hm-cells 为零到五个的患者。因此，hm-cells 的流行率是前列腺癌预后不良的预测因子，也是针对靶向基质成分和防止上皮-间充质转化的药物的潜在预测和治疗反应生物标志物。
