Multi-omics profiling identifies triciribine as a biomarker-guided antimetabolite therapy for acute lymphoblastic leukemia.

Acute lymphoblastic leukemia (ALL) exhibits molecular and metabolic heterogeneity that limits therapeutic efficacy. Although antimetabolites remain central to ALL treatment, determinants of therapeutic sensitivity and resistance remain incompletely understood. Here, we investigated the activity and molecular response landscape of the purine analog triciribine (TCN) across diverse ALL models using integrated phosphoproteomics, thermal proteome profiling, and quantitative proteomics. TCN showed potent cytotoxicity across ALL cell lines and a drug response profile distinct from canonical PI3K-Akt-mTOR inhibitors. Early phosphoproteomics revealed no consistent suppression of Akt pathway signaling, with transient activation of several Akt-associated readouts and inhibition of cyclin-dependent kinase signaling. The monophosphorylated metabolite TCN-P was the predominant intracellular active species, and adenosine kinase (ADK) was required for TCN-P generation and cytotoxicity. Proteome Integral Solubility Alteration profiling identified candidate TCN-P interactions involving nucleotide metabolism, DNA replication, and protein synthesis, associated with impaired purine biosynthesis, DNA damage, translational stress, and cell-cycle perturbation. Across ALL cell lines and primary samples, ADK levels correlated positively with TCN sensitivity, supporting ADK-dependent metabolism as a major determinant of therapeutic response. Together, these findings identify TCN as a metabolically activated antileukemic agent with broad antimetabolite activity and nominate ADK as a potential predictive biomarker for precision therapy in ALL.