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Imaging Mass Cytometry (IMC) as a Tool to Characterize Circulating Tumor Cells (CTCs) in Preclinical Mouse Models

Pore, M.; Balamurugan, K.; Atkinson, A.; Breen, D.; Mallory, P.; Cardamone, A.; McKennett, L.; Newkirk, C.; Sharan, S.; Bocik, W.; Sterneck, E.

2026-04-16 cancer biology
10.64898/2025.12.18.695262 bioRxiv
Show abstract

Circulating tumor cells (CTCs), and especially CTC-clusters, are linked to poor prognosis and may reveal mechanisms of metastasis and treatment resistance. Therefore, developing unbiased methods for the functional characterization of CTCs in liquid biopsies is an urgent need. Here, we present an evaluation of multiplex imaging mass cytometry (IMC) to analyze CTCs in mice with human xenograft tumors. In a single-step process, IMC uses metal-labeled antibodies to simultaneously detect a large number of proteins/modifications within minimally manipulated small volumes of blood from the tail vein or heart. We used breast cancer cell lines and a patient-derived xenograft (PDX) to assess antibodies for cross-species interpretation. Along with manual verification, HALO-AI-based cell segmentation was used to identify CTCs and quantify markers. Despite some limitations regarding human-specificity, this technology can be used to investigate the effect of genetic and pharmacological interventions on the properties of single and cluster CTCs in tumor-bearing mice.

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