Journal Article

Circulating tumor cell detection and single‐cell analysis using an integrated workflow based on ChimeraX®‐i120 Platform: A prospective study

Zhou, Jian, Zhang, Ze‐Fan, Fan, Jia, Cheng, Jian‐Wen, Chen, Li‐Meng, Wang, Peng‐Xiang, Jin, Wei‐Xiang, Zhou, Kai‐Qian, Hu, Bo, Xu, Yang, Guo, Wei, Yang, Xin‐Rong, Cao, Ya, Peng, Hai‐Xiang, Wu, Sui‐Yi, Sun, Yun‐Fan, Huang, Kai

2021

Overview

We developed an integrated workflow for circulating tumor cell (CTC) detection and downstream single‐cell analysis based on a novel ChimeraX®‐i120 platform. The platform facilitates negative enrichment, immunofluorescent labeling, and machine learning‐based identification of CTCs. The CTC captured by the platform is also compatible for single‐cell molecular analysis. In this study, potential utility of our workflow was validated in clinical setting. Circulating tumor cell (CTC) analysis holds great potential to be a noninvasive solution for clinical cancer management. A complete workflow that combined CTC detection and single‐cell molecular analysis is required. We developed the ChimeraX®‐i120 platform to facilitate negative enrichment, immunofluorescent labeling, and machine learning‐based identification of CTCs. Analytical performances were evaluated, and a total of 477 participants were enrolled to validate the clinical feasibility of ChimeraX®‐i120 CTC detection. We analyzed copy number alteration profiles of isolated single cells. The ChimeraX®‐i120 platform had high sensitivity, accuracy, and reproducibility for CTC detection. In clinical samples, an average value of > 60% CTC‐positive rate was found for five cancer types (i.e., liver, biliary duct, breast, colorectal, and lung), while CTCs were rarely identified in blood from healthy donors. In hepatocellular carcinoma patients treated with curative resection, CTC status was significantly associated with tumor characteristics, prognosis, and treatment response (all P < 0.05). Single‐cell sequencing analysis revealed that heterogeneous genomic alteration patterns resided in different cells, patients, and cancers. Our results suggest that the use of this ChimeraX®‐i120 platform and the integrated workflow has validity as a tool for CTC detection and downstream genomic profiling in the clinical setting.

• Publisher: John Wiley & Sons, Inc,John Wiley and Sons Inc,Wiley
• Subject: Blood / Breast cancer / Cancer / Carcinoma, Hepatocellular - blood / Carcinoma, Hepatocellular - diagnosis / Cell adhesion & migration / Cell adhesion molecules / Cell Line, Tumor / Cholangiocarcinoma / Chronic infection / circulating tumor cell / Cirrhosis / Colorectal cancer / Colorectal carcinoma / Copy number / Cytokeratin / Development and progression / Enumeration / Epithelial cells / Erythrocytes / Ethylenediaminetetraacetic acid / Fluorescent Antibody Technique / Genomes / Health aspects / Hepatitis / Hepatitis B / Hepatocellular carcinoma / Humans / Integrated approach / integrated platform / liquid biopsy / Liver cancer / Liver cirrhosis / Liver Neoplasms - blood / Liver Neoplasms - diagnosis / Lung cancer / Machine Learning / machine learning‐based image recognition / Metastases / Metastasis / Neoplasms - blood / Neoplastic Cells, Circulating / Polymerase chain reaction / Population studies / Prospective Studies / Semiconductors / Single-Cell Analysis - methods / single‐cell sequencing / Whole genome sequencing / Workflow / α-Fetoprotein