Network-driven prioritization and functional phenotyping nominate TTC23 as a biomarker-informed target in chlorpromazine repurposing for glioblastoma.

Background: Glioblastoma (GBM) remains a lethal brain tumor with limited therapeutic options and near-universal recurrence. Drug repurposing offers a practical strategy, but pleiotropic compounds require systematic target triage to yield actionable and testable vulnerabilities. Methods: We integrated GBM transcriptomic dysregulation with curated chlorpromazine (CPZ)-associated targets to define drug-disease intersecting genes, constructed a protein-protein interaction network, and developed an outcome-linked Lasso-Cox prognostic model to prioritize core candidates. Structure-informed docking and coarse-grained conformational sampling were used to evaluate the plausibility of a TTC23-CPZ interaction. TTC23-associated pathway activity, oncogenic state features, and immune contexture were characterized using expression stratification, enrichment and state scoring, cancer-immunity cycle analysis, and immune infiltration estimation. Functional validation was performed in GBM cell models to assess migration, apoptosis, cell viability, and clonogenic potential under TTC23 perturbation with or without CPZ exposure. Results: Integrated CPZ-GBM intersection analysis and network-based prognostic modeling consistently prioritized TTC23 as a clinically relevant candidate. Structure-based analyses supported a consistent TTC23-CPZ interaction hypothesis across conformational sampling. Elevated TTC23 expression was associated with coordinated pathway activation, malignant functional states, and distinct immune-associated features. Functionally, TTC23 depletion suppressed migratory capacity, increased apoptotic susceptibility, reduced short-term viability, and impaired long-term clonogenic survival, while sensitizing GBM cells to CPZ-associated anti-tumor phenotypes. Conclusion: Our multi-layer framework nominates TTC23 as a functionally relevant determinant associated with CPZ response in GBM and supports the CPZ-TTC23 axis as a candidate for biomarker-informed drug repurposing.