Abstract
Purpose
Iron acquisition is key to maintaining cell survival and function. Cancer cells in general are considered to have an insatiable iron need. Iron delivery via the transferrin/transferrin receptor pathway has been the canonical iron uptake mechanism. Recently, however, our laboratory and others have explored the ability of ferritin, particularly the H-subunit, to deliver iron to a variety of cell types. Here, we investigate whether Glioblastoma (GBM) initiating cells (GICs), a small population of stem-like cells, are known for their iron addiction and invasive nature acquire exogenous ferritin, as a source of iron. We further assess the functional impact of ferritin uptake on the invasion capacity of the GICs.
Methods
To establish that H-ferritin can bind to human GBM, tissue-binding assays were performed on samples collected at the time of surgery. To interrogate the functional consequences of H-ferritin uptake, we utilized two patient-derived GIC lines. We further describe H-ferritin’s impact on GIC invasion capacity using a 3D invasion assay.
Results
H-ferritin bound to human GBM tissue at the amount of binding was influenced by sex. GIC lines showed uptake of H-ferritin protein via transferrin receptor. FTH1 uptake correlated with a significant decrease in the invasion capacity of the cells. H-ferritin uptake was associated with a significant decrease in the invasion-related protein Rap1A.
Conclusion
These findings indicate that extracellular H-ferritin participates in iron acquisition to GBMs and patient-derived GICs. The functional significance of the increased iron delivery by H-ferritin is a decreased invasion capacity of GICs potentially via reduction of Rap1A protein levels.
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Data availability
All data and methodology supporting the findings of this study are available within the paper and its Supplementary Information files.
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Acknowledgements
We would like to sincerely thank Gonzalo Hernandez Viedma and Dr. Mayka Sánchez Fernández of UIC Barcelona International University of Catalonia for their help in analyzing the mRNA sequences for IRE. We also thank Nate Sheaffer, Joseph Bednarczyk, and Dr. Thomas Abraham from Penn State College of Medicine’s Flow Cytometry and Microscopy cores for their assistance and guidance with flow cytometry and microscopy analysis. The Advanced Light Microscopy core (RRID: SCR_022526) services and instruments used in this project were funded, in part, by the Pennsylvania State University College of Medicine via the Office of the Vice Dean of Research and Graduate Students. We also thank Penn State Laboratory for Isotopes and Metals in the Environment (LIME) for performing ICP-AES on our protein samples. The GBM tissue was obtained from the Penn State Neurosurgery BioRepository.
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This work was funded by National Institutes of Health under Grant P01CA245705. The contents of this paper are solely the responsibility of the authors and do not necessarily represent the official views of the National Institutes of Health.
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Author contributions are as follows. BPS: conceptualization, data curation, formal analysis, investigation, methodology, writing—original draft, and writing—review and editing. BWS: methodology, and human subjects’ data collection and analysis. GS: data curation, formal analysis, and software. VS: data curation, formal analysis, and software. BEZ: human patient tissue resection and collection and patient data collection. JRC: conceptualization, funding acquisition, supervision, and writing—review and editing. All authors reviewed the manuscript.
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Human GBM tissues used in the study were collected following an institutional review board (IRB)-compliant protocol. (Penn State Hershey Neuroscience Institute Biorepository IRB #2914, approved). Informed consents were obtained from all individual participants included in the study.
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(Pandya) Shesh, B., Slagle-Webb, B., Shenoy, G. et al. Uptake of H-ferritin by Glioblastoma stem cells and its impact on their invasion capacity. J Cancer Res Clin Oncol 149, 9691–9703 (2023). https://doi.org/10.1007/s00432-023-04864-2
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DOI: https://doi.org/10.1007/s00432-023-04864-2