Hybrid Computational Modeling with Multi-Level Validation Identifies TK1-VIM as a Robust Therapeutic Pair in...

Triple-negative breast cancer (TNBC) lacks effective molecular targets, leading to poor prognosis. Previous computational methods to identify targets have suffered from low druggability, high complexity, and lack of robust validation. We propose a hybrid methodology combining Boolean network modeling with semidefinite programming (SDP) to analyze a TNBC cell line network. The resulting therapeutic pair underwent a multi-level validation framework, including Boolean simulations, statistical uncertainty quantification (bootstrap), sensitivity analysis, and orthogonal computational support from AlphaGenome, a deep learning model from Google DeepMind. Our analysis identified TK1 and VIM as a computationally robust therapeutic pair. Dual inhibition achieved 99.03% similarity to the apoptotic state with a 95% confidence interval of [98.79%, 99.26%], and was statistically superior to alternative pairs (p<0.001). The selection remained optimal across all tested model parameters, demonstrating high robustness. Importantly, the pair has full druggability because both targets have available specific inhibitors. Orthogonal computational evidence from AlphaGenome, stratified by mammary compartment, indicated that both targets exhibit moderate baseline expression in normal mammary epithelium (TK1 = 0.159, VIM = 0.143 in normalized RNA-seq units; n = 13 tracks per gene), with VIM showing a 2.2-fold higher expression in mammary stroma than in epithelium-a gradient consistent with its established role as a mesenchymal marker. Promoter-variant proxy analysis indicated near-zero transcriptomic perturbation upon simulated inhibition of either target in normal mammary epithelium (mean |log2FC|<0.001), supporting a favorable therapeutic window. Our methodology identified TK1-VIM as a computationally robust, druggable therapeutic candidate pair with biologically plausible mechanism of action. Gene-variability analysis identified TK1 and VIM as the highest-scoring candidates, with SDP optimization providing complementary, independent confirmation of this selection. This work provides a computationally grounded candidate strategy and a rigorous methodological benchmark for computational drug target identification; experimental validation remains an essential next step before clinical translation.