CAS Key Laboratory of Standardization and Measurement for Nanotechnology, CAS Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety, CAS Center for Excellence in Nanoscience, National Center for Nanoscience and Technology of China, Beijing 100190, China.
Lab Chip. 2019 Sep 27;19(19):3168-3178. doi: 10.1039/c9lc00473d.
Whole-genome sequencing on circulating tumor cells (CTCs) at the single cell level has recently been found helpful for precision medicine, as the oncogenic profiles of single CTCs are useful for discovering oncogenic mutation heterogeneities and guiding/adjusting cancer treatment. To overcome the limits of existing methods of single CTC sequencing, in which CTC enrichment, identification and gene amplification are performed by discrete modules, this study presents a novel method in which all processing steps from blood sample collection to preparation of gene amplification products for sequencers are finished in a single microfluidic chip. This microfluidic chip comprehensively performs blood filtering, CTC enrichment, CTC identification/isolation, CTC lysis and whole genome amplification (WGA) at the single cell level. By sequencing single CTCs from clinical blood samples with pointing key driver and drug-resistance mutations, the novel microfluidic chip was validated to be capable of genetically profiling single CTCs with minimum cell loss/human labor, and more importantly, high accuracy and repeatability, which are crucial factors for promoting clinical application of single CTC sequencing.
循环肿瘤细胞（CTC）的全基因组测序最近在精准医学方面得到了应用，因为单个 CTC 的致癌谱对于发现致癌突变异质性和指导/调整癌症治疗很有用。为了克服现有单细胞 CTC 测序方法的局限性，这些方法通过离散模块来进行 CTC 富集、鉴定和基因扩增，本研究提出了一种新方法，该方法将从血液样本采集到测序仪基因扩增产物制备的所有处理步骤都在单个微流控芯片中完成。该微流控芯片在单细胞水平上全面进行血液过滤、CTC 富集、CTC 鉴定/分离、CTC 裂解和全基因组扩增（WGA）。通过对临床血液样本中的单个 CTC 进行测序，以确定关键驱动和耐药性突变，验证了新型微流控芯片能够以最小的细胞损失/人力对单个 CTC 进行基因分析，更重要的是，具有高精度和可重复性，这对于促进单细胞 CTC 测序的临床应用至关重要。
