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5-Fluorouracil Induces an Acute Reduction in Neurogenesis and Persistent Neuroinflammation in a Mouse Model of the Neuropsychological Complications of Chemotherapy

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Abstract

The neuropsychological symptoms associated with chemotherapy treatment remain a major challenge with their prevention hampered by insufficient understanding of pathophysiology. While long-term neuroimmune changes have been identified as a hallmark feature shared by neurological symptoms, the exact timeline of mechanistic events preceding neuroinflammation, and the relationship between the glial cells driving this neuroinflammatory response, remain unclear. We therefore aimed to longitudinally characterize the neuroimmunological changes following systemic 5-fluorouracil (5-FU) treatment to gain insight into the timeline of events preceding the well-documented chronic neuroinflammation seen following chemotherapy. Eighteen female C57Bl/6 mice received a single intraperitoneal dose of 5-FU and groups were killed at days 1 and 2 (acute timepoint), days 4 and 8 (subacute timepoint), and days 16 and 32 (chronic timepoint). A further six mice were administered with vehicle control with tissues collected from three mice on day 1 and day 32 of the study. The expression of key genes of interest, BCL2, BDNF, TIMP1, MMP-9, MMP-2, TNFα, IL-1β, and IL-6R were assessed using real time polymerase chain reaction. Levels of neurogenesis were determined through immunofluorescent staining of doublecortin (DCX). The density of microglia and astrocytes were assessed using immunofluorescence staining of Iba1 and GFAP respectively. 5-FU treatment caused significant decreases to DCX staining at acute timepoints (p = 0.0030) which was positively correlated with BCL2 expression levels. An increase to microglial density was observed in the prefrontal cortex (p = 0.0256), CA3 region (p = 0.0283), and dentate gyrus (p = 0.0052) of the hippocampus at acute timepoints. 5-FU caused increases to astrocyte density, across multiple brains regions, at subacute and chronic timepoints which were positively correlated with TNFα, TIMP-1, MMP-2, and IL-6R expression. This study has identified acute objective neuroinflammatory changes suggesting that the role of early intervention should be explored to prevent the development of neuropsychological deficits in the longer-term following chemotherapy.

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Data Availability

The datasets generated within this study are not currently available to the public. However, can be made available upon request to the corresponding author.

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Acknowledgements

Ines Semendric for providing technical assistance. Adelaide Microscopy, University of Adelaide and Bioresources Staff, SAHMRI.

Funding

This work was supported by a Research Training Program Australia Stipend, The Doctor Chun Chung Wong and Madam So Sau Lam Memorial Postgraduate Cancer Research Top-Up Scholarship (CBS), and an NHMRC CJ Martin Biomedical Research Fellowship (HRW).

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Authors and Affiliations

  1. School of Biomedicine, Adelaide Medical School, University of Adelaide, Adelaide, South Australia, Australia

    Courtney B. Subramaniam, Vivien Heng, Marc A. Gladman & Joanne M. Bowen

  2. Supportive Oncology Research Group, Precision Medicine (Cancer), South Australian Health and Medical Research Institute (SAHMRI), University of Adelaide, Adelaide, South Australia, Australia

    Courtney B. Subramaniam, Hannah R. Wardill & Maya R. Davies

Authors
  1. Courtney B. Subramaniam
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  2. Hannah R. Wardill
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  4. Vivien Heng
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  5. Marc A. Gladman
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  6. Joanne M. Bowen
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Contributions

Courtney Subramaniam: methodology, investigation, formal analysis, writing – original draft; Hannah Wardill: supervision, conceptualisation, formal analysis, writing – review & editing; Maya Davies: investigation; Vivien Heng: investigation; Marc Gladman: supervision, writing – review & editing; Joanne Bowen: supervision, conceptualization, writing – review & editing.

Corresponding author

Correspondence to Courtney B. Subramaniam.

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Ethics Approval

The study was approved by the Animal Ethics Committee of the South Australian Health and Medical Research Institute (SAHMRI, #SAM20-034) and complied with the National Health and Research Council (Australia) Code of Practice for Animal Care in Research and Training (2014).

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Competing Interests

Joanne Bowen has received research funding from AstraZeneca, Helsinn Healthcare, Pfizer Pharmaceuticals, PanTheryx, and Puma Biotechnology Inc. Hannah Wardill has received research funding from Nutricia (Danone) Research. Remaining authors have no conflicts of interest to declare.

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Subramaniam, C.B., Wardill, H.R., Davies, M.R. et al. 5-Fluorouracil Induces an Acute Reduction in Neurogenesis and Persistent Neuroinflammation in a Mouse Model of the Neuropsychological Complications of Chemotherapy. Mol Neurobiol 60, 1408–1424 (2023). https://doi.org/10.1007/s12035-022-03136-3

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