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    Area of Science:

    • Oncology
    • Molecular Biology
    • Clinical Diagnostics

    Background:

    • Locally advanced rectal cancer (LARC) treatment involves neoadjuvant chemoradiation therapy (nCRT), but patient response varies significantly.
    • Identifying non-responders to nCRT is crucial to avoid ineffective treatment and associated toxicities.
    • Personalized medicine approaches are needed to optimize treatment selection for LARC patients.

    Purpose of the Study:

    • To develop and validate a predictive assay for nCRT response in LARC patients.
    • To stratify LARC patients based on DNA damage response (DDR) proteins in peripheral blood monocytes (PBMCs).
    • To establish a framework for predicting nCRT response using a novel blood assay.

    Main Methods:

    • Collected PBMCs from LARC patients before and after nCRT, categorized by neoadjuvant rectal (NAR) score.
    • Assessed DDR using immunofluorescence (γH2AX foci) and Luminex xMAP assay for multiple DDR proteins (Chk1, Chk2, γH2AX, p53, ATR, MDM2, p21).
    • Validated assay performance and analyzed clinical associations between DDR protein levels and treatment response.

    Main Results:

    • PBMCs from poor responders showed significantly lower γH2AX foci compared to complete responders (p<0.0001).
    • The xMAP assay demonstrated high precision, reproducibility, and linearity.
    • Six DDR proteins (ATR, MDM2, Chk1, Chk2, γH2AX, p53) significantly differentiated complete responders from poor responders (p ≤ 1e-5).

    Conclusions:

    • An optimized assay can assess DDR proteins in PBMCs to predict nCRT response in LARC patients.
    • Specific protein thresholds identified accurately segregate responders from non-responders.
    • This predictive assay supports personalized oncology by optimizing treatment efficacy and minimizing toxicity.