Phospho 394Y LCK flow-cytometry readout predicts dasatinib sensitivity in paediatric T-cell acute lymphoblastic leukaemia

Abstract

Children with T-cell acute lymphoblastic leukaemia (T-ALL) who relapse or fail induction have poor outcomes. A subset of cases shows glucocorticoid resistance reversible by dasatinib through inhibition of LCK-dependent signalling. To identify a practical biomarker of dasatinib response, we compared in-vitro drug sensitivity with basal phosphorylation of pre-TCR pathway proteins in 28 paediatric T-ALL patient-derived xenografts (PDX). Phospho-flow cytometry quantified LCK (pY394), ZAP70 (pY319) and CD3{zeta} (pY142), normalised to internal controls. Dasatinib IC50 values correlated significantly with pLCK and pZAP70, with pLCK providing the best single-marker performance across clinically relevant thresholds. Logistic, ROC and precision-recall analyses confirmed that pLCK alone achieved excellent classification (AUC [≥] 0.9), while multivariate models added minimal predictive value. Model selection using LASSO and Bayesian Information Criterion further supported pLCK as the dominant predictor. These findings establish pLCK as a robust, scalable biomarker of dasatinib sensitivity suitable for diagnostic integration. A multicentre international validation programme is underway to harmonise pLCK assay protocols, expand testing across biobanks, and assess clinical feasibility in newly diagnosed and relapsed patients within the ALLTogether and HEM-iSMART trials respectively.