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Sexually dimorphic effect of H-ferritin genetic manipulation on survival and tumor microenvironment in a mouse model of glioblastoma

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Abstract

Purpose

Iron plays a crucial role in various biological mechanisms and has been found to promote tumor growth. Recent research has shown that the H-ferritin (FTH1) protein, traditionally recognized as an essential iron storage protein, can transport iron to GBM cancer stem cells, reducing their invasion activity. Moreover, the binding of extracellular FTH1 to human GBM tissues, and brain iron delivery in general, has been found to have a sex bias. These observations raise questions, addressed in this study, about whether H-ferritin levels extrinsic to the tumor can affect tumor cell pathways and if this impact is sex-specific.

Methods

To interrogate the role of systemic H-ferritin in GBM we introduce a mouse model in which H-ferritin levels are genetically manipulated. Mice that were genetically manipulated to be heterozygous for H-ferritin (Fth1+/-) gene expression were orthotopically implanted with a mouse GBM cell line (GL261). Littermate Fth1 +/+ mice were used as controls. The animals were evaluated for survival and the tumors were subjected to RNA sequencing protocols. We analyzed the resulting data utilizing the murine Microenvironment Cell Population (mMCP) method for in silico immune deconvolution. mMCP analysis estimates the abundance of tissue infiltrating immune and stromal populations based on cell-specific gene expression signatures.

Results

There was a clear sex bias in survival. Female Fth1+/- mice had significantly poorer survival than control females (Fth1+/+). The Fth1 genetic status did not affect survival in males. The mMCP analysis revealed a significant reduction in T cells and CD8 + T cell infiltration in the tumors of females with Fth1+/- background as compared to the Fth1+/+. Mast and fibroblast cell infiltration was increased in females and males with Fth1+/- background, respectively, compared to Fth1+/+ mice.

Conclusion

Genetic manipulation of Fth1 which leads to reduced systemic levels of FTH1 protein had a sexually dimorphic impact on survival. Fth1 heterozygosity significantly worsened survival in females but did not affect survival in male GBMs. Furthermore, the genetic manipulation of Fth1 significantly affected tumor infiltration of T-cells, CD8 + T cells, fibroblasts, and mast cells in a sexually dimorphic manner. These results demonstrate a role for FTH1 and presumably iron status in establishing the tumor cellular landscape that ultimately impacts survival and further reveals a sex bias that may inform the population studies showing a sex effect on the prevalence of brain tumors.

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Data availability

All data and methodology supporting the findings of this study are available within the paper. The RNA sequencing data is deposited at NCBI Gene Expression Omnibus repository.

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Acknowledgements

We sincerely thank Penn State College of Medicine’s Genomic Sciences Core members, Dr. Sirisha Pochareddy and Oana Bolt, for their assistance in tissue RNA isolation and mRNA sequencing-related efforts.

Funding

This work was funded by grant P01CA245705 by the National Institutes of Health. 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 (NIH).

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Author notes

  1. Bhavyata Pandya Shesh and Vonn Walter are co - lead authors.

Authors and Affiliations

  1. Department of Neurosurgery, Penn State College of Medicine, Hershey, PA, USA

    Bhavyata Pandya Shesh, Kondaiah Palsa, Becky Slagle-Webb, Elizabeth Neely & James R. Connor

  2. Department of Public Health Sciences, Penn State College of Medicine, Hershey, PA, USA

    Vonn Walter

  3. Department of Microbiology and Immunology, Penn State College of Medicine, Hershey, PA, USA

    Todd Schell

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  1. Bhavyata Pandya Shesh
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Contributions

Conceptualization: Bhavyata (Pandya) Shesh, James R. Connor. Methodology: Bhavyata (Pandya) Shesh, Becky Slagle- Webb, Elizabeth Neely, Vonn Walter, Kondaiah Palsa, Todd Schell. Formal analysis and investigation: Bhavyata (Pandya) Shesh, Vonn Walter. Writing - original draft preparation: Bhavyata (Pandya) Shesh. Writing - review and editing: Bhavyata (Pandya) Shesh, James R. Connor, Vonn Walter. Funding acquisition: James R. Connor. Resources: James R. Connor, Vonn Walter, Todd Schell. Supervision: James R. Connor.

Corresponding author

Correspondence to James R. Connor.

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The authors have no relevant financial or non-financial interests to disclose.

Ethics approval

Mice used in this study were handled following an Institutional Animal Care and Use Committee (IACUC) - compliant protocol. Penn State Hershey College of Medicine IACUC # PROTO202001393 approved.

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Pandya Shesh, B., Walter, V., Palsa, K. et al. Sexually dimorphic effect of H-ferritin genetic manipulation on survival and tumor microenvironment in a mouse model of glioblastoma. J Neurooncol 164, 569–586 (2023). https://doi.org/10.1007/s11060-023-04415-2

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