Synthetic Lethality via Multi-Node Metabolic Locking: A p53-Independent Strategy to Overcome Cisplatin Resistance in Lung Adenocarcinoma

Chemoresistance in lung adenocarcinoma remains a formidable challenge, particularly in phenotypes harboring p53 mutations that bypass conventional apoptotic pathways. Metabolic plasticity allows these cells to navigate therapeutic stress, necessitating a transition from single-target inhibition to systemic network collapse. Herein, we employed a systems biology framework using the Human-GEM (v1.18.0) genome-scale metabolic model to investigate the synergistic effects of Vitamin D3, Vitamin K2, and Genistein. Simulations were performed on A549 (p53-wildtype) and H1299 (p53-null) cell lines under nutrient-constrained media conditions. We utilized Flux Balance Analysis (FBA) and Flux Variability Analysis (FVA) to quantify metabolic solution spaces and identify lethal bottlenecks. Preliminary in vitro cytotoxicity assays were conducted to establish a safe therapeutic window. Our results identified values for Vitamin D3 and K2 at 45 and 18 µM, respectively. Subsequent combination treatments at sub-toxic concentrations showed no significant decrease in cell viability (p=0.286) or PAK1 protein expression, confirming that the synergistic metabolic lock functions through flux redistribution rather than acute proteomic degradation or non-specific toxicity. Our simulations reveal that the tripartite synergy induces a state of Metabolic Exhaustion by simultaneously locking the glycolytic exit (PKM/HK2) and depleting the antioxidant reservoir via Pentose Phosphate Pathway (PPP) restriction. While single-agent treatments showed minimal impact, the combined intervention achieved a 100% biomass reduction in the aggressive p53-null H1299 model. This synthetic lethality was achieved independently of the p53 status, demonstrating that targeting the PAK1/CYP24A1-mediated metabolic rheostat effectively eliminates all viable bypass pathways for survival and DNA repair. Conclusions: We provide a computational blueprint for a universal Metabolic Lock strategy that re-sensitizes resistant lung cancer cells to Cisplatin. By mathematically proving the transition from metabolic plasticity to total exhaustion, this study underscores the potential of rational nutrient-based adjuvants to overcome genetic heterogeneity and chemoresistance in clinical oncology.