Single-Cell Multi-Dataset Integration Reveals Mitoxyperilysis As A Conserved Temporal-Spatial Cell Death Mechanism In Cisplatin-Induced Ototoxicity
摘要
Background: Cisplatin-induced ototoxicity (CIO) causes irreversible sensorineural hearing loss in 40–80% of treated patients, yet no FDA-approved otoprotectant currently exists to prevent this dose-limiting toxicity. Mitoxyperilysis—mitochondrial oxidative membrane rupture governed by BAX, BAK1, BID, and mTORC2 (Wang et al., Cell 2025)—is a novel, mechanistically distinct form of regulated cell death whose role in cochlear injury has not previously been characterised.
Methods: Three GEO datasets (GSE136196, GSE165662, GSE281324) comprising 9,678 stria vascularis (SV) cells across four experimental conditions (adult chronic cisplatin/control; P6 acute cisplatin/control) were integrated employing Harmony batch-effect correction and UMAP dimensionality reduction. Mitoxyperilysis pathway scores were computed per cell. Differential expression analysis, followed by LASSO-regularised logistic regression, Random Forest classification (n_estimators=500; 5-fold cross-validation), and WGCNA co-expression network analysis, were subsequently applied to identify 20 SV hub genes. Network pharmacology queries against DrugBank v5.1.11 and STITCH v5.0 prioritised otoprotective candidates. Structural validation of the dominant hub gene was conducted via 100 ns all-atom molecular dynamics (MD) simulation (CHARMM36m force field; POPC/POPE bilayer) and AutoDock Vina 2026 molecular docking.
Results: The integrated SV atlas comprised 9,678 cells annotated into four subpopulations: Marginal Cells (n=3,064), Intermediate Cells (n=2,055), Basal Cells (n=2,308), and Fibrocytes (n=2,251). Adult SV Mitoxyperilysis scores declined from Ctrl_Chronic (0.447) to Cis_Chronic (0.266), reflecting survivor-bias depletion of pathway-executing cells. mt-Nd4l ranked first in Random Forest feature importance (importance=0.0731), and the prognostic model achieved AUC=0.938 (Random Forest) and AUC=0.921 (LASSO-LR). Marginal Cells exhibited the highest median cisplatin risk score (~0.65). Network pharmacology prioritised Rifampicin, Rapamycin, and Idebenone as otoprotective candidates; Idebenone provided dual targeting of MT-ND4L and COX20. MD simulation confirmed transmembrane helical stability (backbone RMSD=6.42±0.83 Å; Rg=23.76±0.75 Å). Idebenone docking demonstrated binding at the Complex I/ND4L membrane interface (ΔG=−4.007 kcal/mol).
Conclusions: This study presents the first computational application of Mitoxyperilysis to CIO, integrating multi-dataset single-cell transcriptomics with machine learning, network pharmacology, and structural biology. Collectively, the findings provide a mechanistic framework centred on mt-Nd4l and the respiratory chain for otoprotective drug development, with Rifampicin, Rapamycin, and Idebenone identified as prioritised candidates warranting experimental validation.
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