Hepatocellular carcinoma (HCC) is a prevalent malignancy whose circulating tumor cells (CTCs) exhibit significant phenotypic heterogeneity and diverse gene expression profiles due to epithelial-mesenchymal transition (EMT). However, current detection methods lack the capacity for simultaneous quantification of multidimensional biomarkers, limiting comprehensive understanding of tumor biology and dynamic changes. The precise quantitative analysis of multi-dimensional molecular characteristics of HCC CTCs has become a focal point in current research.
Recently, the research team led by Professors Lianqun Zhou at Suzhou Institute of Biomedical Engineering and Technology (SIBET) of the Chinese Academy of Sciences developed an innovative CTC Digital Simultaneous Cross-dimensional Output and Unified Tracking (d-SCOUT) technology, as shown in Figure 1. Based on multi-channel radial cross-flow CTC Chip (MRX-CTC chip) and multi-real-time digital PCR (MRT-dPCR), this platform efficiently enriches CTCs while preserving protein and mRNA integrity. Specifically, the researchers converted CTC protein detection into nucleic acid detection using immuno-PCR principles, achieving unified quantification of proteins (ASGPR, GPC-3, and EpCAM) and mRNAs (PD-L1, GPC-3, and EpCAM). The technology demonstrates excellent sensitivity (LOD of 3.2 CTCs per mL of blood) and reproducibility (mean %CV=1.80-6.05%).
In a clinical study involving 99 samples, molecular signatures derived from HCC CTCs showed strong diagnostic potential (AUC=0.950, sensitivity=90.6%, specificity=87.5%), as illustrated in Figure 2. Moreover, by integrating machine learning, d-SCOUT enables clustering of CTC characteristics at both mRNA and protein levels, mapping normalized heterogeneous two-dimensional molecular profiles to assess HCC metastatic risk (Figure 3). Dynamic digital tracking of eight HCC patients undergoing different treatments visually demonstrated therapeutic effects, validating the technology's capability to quantify treatment efficacy (Figure 3).
The research, titled "Digital Quantitative Detection for Heterogeneous Protein and mRNA Expression Patterns in Circulating Tumor Cells," was published in the journal Advanced Science. Researchers Chuanyu Li, Wei Zhang, and Lianqun Zhou from the SIBET served as the co-corresponding authors, while Hao Li and Jinze Li were the co-first authors. This study was supported by the National Key R&D Program, the National Natural Science Foundation of China, and the Youth Innovation Promotion Association of the Chinese Academy of Sciences.
Figure 1 Simultaneous acquisition and quantification of HCC CTC
transcriptotypic andphenotypic biomarkers for personalized HCC management
Figure 2 D-SCOUT assays for simultaneous quantification of HCC CTC
transcriptotypes and phenotypes across different cohorts
Figure 3 Multidimensional digital molecular information profiling of HCC CTCs
via d-SCOUT analysis
