Decoding cytarabine-induced damage in the prepubertal testis: A multi-omics atlas of ecosystem and inheritance

Pediatric leukemia therapies can compromise future fertility, particularly in prepubertal boys who cannot bank sperm. We used dose-response histology, endocrine and sperm analyses, bulk and single-cell RNA-seq, and metabolomics to map cytarabine (Ara-C) injury in prepubertal mouse testes. Ara-C caused seminiferous-tubule disruption, hormone suppression, sperm loss, spermatogonial depletion linked to ferroptosis and replication arrest, stress remodeling of Sertoli and Leydig cells, and altered intercellular signaling centered on Sertoli hubs. Unexposed F1 offspring retained inflammatory, nucleotide, and energy-metabolism changes, suggesting intergenerational imprinting after paternal exposure. These findings define cell-type-specific vulnerabilities in the developing testis and nominate redox balance, ferroptosis, Sertoli stress pathways, Leydig inflammatory signaling, and niche communication as targets for fertility-preserving strategies during pediatric chemotherapy.