Integrated transcriptomic profiling identifies a clinically adverse iron-autophagy imbalance state in neuroblastoma
Year: 2026
Type: article
Abstract: Background: Neuroblastoma is characterized by marked molecular heterogeneity, and additional biological states contributing to aggressive disease remain incompletely defined. Ferritinophagy-related genes connect ferritin turnover, intracellular iron regulation, and autophagy-associated processes, but whether they delineate clinically relevant iron-autophagy transcriptional states in neuroblastoma is unclear. Methods: We integrated bulk transcriptomic, single-cell RNA-sequencing, and region-of-interest (ROI)-level GeoMx Digital Spatial Profiling datasets. Ferritinophagy-related genes were used as an entry point for unsupervised classification in the GSE62564 discovery cohort, followed by construction of a locked 34-gene centroid classifier and a continuous C1-associated axis, which were projected without gene reselection or refitting into three independent bulk cohorts. A four-gene iron-autophagy imbalance index (IA4) was derived from opposing iron-handling and autophagy-associated transcriptional components. Single-cell and spatial analyses assessed malignant-cell expression, treatment-associated differences within malignant cells, and clinical-risk associations. Results: Two transcriptional states were identified. In GSE62564, C1 comprised 180 of 498 tumors and was associated with inferior overall survival (OS) [univariable hazard ratio (HR), 16.51; 95% confidence interval (CI): 9.52–28.63] and event-free survival (EFS) (HR, 4.24; 95% CI: 3.13–5.74); at 5 years, C1 was associated with a 1.45-year loss of restricted mean OS (95% CI: 1.18–1.72). The associations remained significant after adjustment for age, stage, and MYCN status (OS HR, 5.03; 95% CI: 2.53–9.99; EFS HR, 2.00; 95% CI: 1.30–3.08). In three independent bulk cohorts, a 1-standard deviation (SD) increase in the continuous C1 score was associated with inferior OS (HR range, 1.58–2.44; all P<0.001). C1 tumors showed coordinated malignant and iron-autophagy-associated programs. IA4, defined from TFRC , PCBP1 , PINK1 , and ATG16L1 , correlated with the continuous C1 axis in 34 pretreatment tumor ROIs (Spearman rho =0.64, P=4.3×10 −5 ) and remained robust in leave-one-gene-out analyses. In 22 paired longitudinal donors, the malignant-cell weighted C1 score decreased after treatment (P=0.002), and the cycle-residual component remained lower after adjustment for S-phase and G2/M activity (P=0.01). Conclusions: Using ferritinophagy-related genes as a discovery framework, this study identifies a clinically adverse iron-autophagy imbalance state in neuroblastoma and provides a compact transcriptional representation through IA4. The findings establish a framework for future functional investigation without implying direct measurement of ferritinophagic flux, intracellular iron, or ferroptosis sensitivity.
Source: Translational Pediatrics
Language: English
Subfield: Oncology
Field: Medicine
Domain: Health Sciences
Open Access status: diamond
OpenAlex ID: W7223694860