Mishra Avanish, Huang Shih-Bo, Dubash Taronish, Burr Risa, Edd Jon F, Wittner Ben S, Cunneely Quinn E, Putaturo Victor R, Deshpande Akansha, Antmen Ezgi, Gopinathan Kaustav A, Otani Keisuke, Miyazawa Yoshiyuki, Kwak Ji Eun, Guay Sara Y, Kelly Justin, Walsh John, Nieman Linda T, Galler Isabella, Chan PuiYee, Lawrence Michael S, Sullivan Ryan J, Bardia Aditya, Micalizzi Douglas S, Sequist Lecia V, Lee Richard J, Franses Joseph W, Ting David T, Brunker Patricia A R, Maheswaran Shyamala, Miyamoto David T, Haber Daniel A, Toner Mehmet
Center for Engineering in Medicine and Surgery, Massachusetts General Hospital and Harvard Medical School, Boston, MA, 02114, USA.
Krantz Family Center for Cancer Research, Massachusetts General Hospital Cancer Center and Harvard Medical School, Charlestown, MA, 02129, USA.
Nat Commun. 2025 Jan 2;16(1):32. doi: 10.1038/s41467-024-55140-x.
Circulating Tumor Cells (CTCs) in blood encompass DNA, RNA, and protein biomarkers, but clinical utility is limited by their rarity. To enable tumor epitope-agnostic interrogation of large blood volumes, we developed a high-throughput microfluidic device, depleting hematopoietic cells through high-flow channels and force-amplifying magnetic lenses. Here, we apply this technology to analyze patient-derived leukapheresis products, interrogating a mean blood volume of 5.83 liters from seven patients with metastatic cancer. High CTC yields (mean 10,057 CTCs per patient; range 100 to 58,125) reveal considerable intra-patient heterogeneity. CTC size varies within patients, with 67% overlapping in diameter with WBCs. Paired single-cell DNA and RNA sequencing identifies subclonal patterns of aneuploidy and distinct signaling pathways within CTCs. In prostate cancers, a subpopulation of small aneuploid cells lacking epithelial markers is enriched for neuroendocrine signatures. Pooling of CNV-confirmed CTCs enables whole exome sequencing with high mutant allele fractions. High-throughput CTC enrichment thus enables cell-based liquid biopsy for comprehensive monitoring of cancer.
血液中的循环肿瘤细胞（CTC）包含DNA、RNA和蛋白质生物标志物，但其临床应用因细胞稀有而受到限制。为了能够对大量血液进行与肿瘤表位无关的检测，我们开发了一种高通量微流控装置，通过高流量通道和力放大磁透镜去除造血细胞。在此，我们应用这项技术分析患者来源的白细胞分离产品，检测了7例转移性癌症患者平均5.83升的血量。高CTC产量（平均每位患者10,057个CTC；范围为100至58,125个）揭示了患者内部存在显著的异质性。患者体内CTC大小各异，67%的CTC直径与白细胞重叠。配对的单细胞DNA和RNA测序确定了CTC内非整倍体的亚克隆模式以及不同的信号通路。在前列腺癌中，缺乏上皮标志物的小非整倍体细胞亚群富含神经内分泌特征。对经拷贝数变异（CNV）确认的CTC进行合并，能够进行具有高突变等位基因分数的全外显子组测序。因此，高通量CTC富集技术能够实现基于细胞的液体活检，用于癌症的全面监测。
