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Cytometry by Time-of-Flight for Profiling Therapeutic Response in Colorectal Cancer: Strengths, Limitations, and
Eleandros Kyros1, Adam Mylonakis2, Pagona Kastanaki1
1First Department of Surgery, National and Kapodistrian University of Athens, Laikon General Hospital, Athens, Greece.
Anticancer Research
|April 28, 2026
Summary
Cytometry by time-of-flight (CyTOF) and imaging mass cytometry (IMC) reveal immune and stromal biomarkers in colorectal cancer (CRC) that predict treatment response. These advanced techniques offer insights beyond traditional imaging for personalized CRC therapy.
Area of Science:
- Single-cell proteomics and spatial profiling in oncology.
- Advancements in colorectal cancer (CRC) research.
Background:
- Current colorectal cancer (CRC) response assessment relies on anatomic imaging, missing dynamic immune and stromal changes.
- High-dimensional single-cell proteomic and spatial profiling techniques like CyTOF and IMC enhance understanding of CRC therapeutic responses.
Purpose of the Study:
- To review current evidence on the application of CyTOF and IMC in characterizing therapeutic response in CRC.
- To summarize findings from human, preclinical, and organoid studies.
Main Methods:
- Systematic literature search of publications using CyTOF or IMC in CRC.
- Prioritization of studies linking single-cell features to treatment response or pharmacodynamics.
- Organization of findings by biological theme and clinical applicability.
Main Results:
- Spatial analyses identified macrophage-T-cell niches (e.g., CD68+, CD74+, C1QC+) predicting PD-1 blockade benefit.
- Cancer-associated fibroblast (CAF)-dense matrices and granulocytic proximity correlate with resistance.
- Systemic CyTOF revealed peri-operative lymphopenia, persistent HLA-DRlow monocytes, and chemotherapy-induced NK cell depletion.
- Organoid studies linked cellular differentiation to ERK activation and therapy tolerance.
Conclusions:
- CyTOF and IMC offer actionable biomarkers (e.g., C1QC+ macrophage-T-cell proximity, HLA-DRlow monocytes, NK cell maturation) for refining CRC response assessment.
- These biomarkers provide insights beyond conventional imaging criteria.
- Harmonization, validation, and clinical integration are necessary for practical application.
