Divergent immediate and delayed effects of juvenile exposure to doxorubicin on the thymus in C57BL/6 mice

Abstract The long-term effects of doxorubicin (DOX) chemotherapy on thymic immune function in childhood cancer survivors remain inadequately understood. This study explores the immediate and delayed impacts of low-dose DOX on thymic immune populations using a juvenile mouse model. Male mice received...

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Main Authors: Benu George, Korbyn J. V. Dahlquist, Marianne K. O. Grant, Mary R. Daniel, Declan M. Smith, Ian Ahlberg, Karim T. Sadak, Davis Seelig, Christina D. Camell, Beshay N. Zordoky
Format: Article
Language:English
Published: Nature Portfolio 2025-05-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-98617-5
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Summary:Abstract The long-term effects of doxorubicin (DOX) chemotherapy on thymic immune function in childhood cancer survivors remain inadequately understood. This study explores the immediate and delayed impacts of low-dose DOX on thymic immune populations using a juvenile mouse model. Male mice received intraperitoneal DOX injections (4 mg/kg/week) for three weeks, with evaluations performed at one- and five-weeks post treatment. Thymic samples were collected and analyzed using multi-parameter flow cytometry to assess changes in immune cell composition and phenotype. Additionally, real-time polymerase chain reaction (RT-PCR) was employed to measure gene expression of cytokines and senescence markers. One week after DOX administration, significant thymic atrophy was evident. While mature CD3+CD4+ T-cell frequency remained unchanged, CD3+CD8+ T-cells significantly increased, suggesting differential effects on T-cell subsets. PD1+ expression increased across naïve and memory CD4+ T-cell subsets, suggesting activation or exhaustion. Additionally, Ki67+ expression was elevated in naïve and memory CD8+ T-cells, indicating enhanced proliferation. Gene expression analysis revealed upregulation of Foxn1, Pax1, Ifnγ, and Il7 whereas Il6 and Il17 were downregulated. Furthermore, Cdkn1a (p21 ) expression was elevated, suggesting immune dysregulation and early immunosenescence. At five weeks, thymic weight rebounded; however, T-cell subsets displayed persistent perturbations. Central memory and effector memory CD4+ T-cells were reduced, while naïve CD4+ T-cells showed increased Ki67+ expression. In contrast, CD8+ T-cells subsets remained largely unchanged, except for a decrease in central memory cells. Although expression of thymus-related genes was normalized, p21 expression remained elevated, suggesting lingering immunosenescence. These findings highlight the complex effects of DOX, including acute thymic atrophy due to T-cell depletion, and a delayed recovery with persistent immunosenescence, underscoring the need for strategies to preserve immune function in childhood cancer survivors.
ISSN:2045-2322