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Cellular and Molecular Biology

PTEN-restoration abrogates brain colonisation and perivascular niche invasion by melanoma cells

British Journal of Cancer volume 130, pages 555–567 (2024)Cite this article

Subjects

Abstract

Background

Melanoma brain metastases (MBM) continue to be a significant clinical problem with limited treatment options. Highly invasive melanoma cells migrate along the vasculature and perivascular cells may contribute to residual disease and recurrence. PTEN loss and hyperactivation of AKT occur in MBM; however, a role for PTEN/AKT in perivascular invasion has not been described.

Methods

We used in vivo intracranial injections of murine melanoma and bulk RNA sequencing of melanoma cells co-cultured with brain endothelial cells (brECs) to investigate brain colonisation and perivascular invasion.

Results

We found that PTEN-null melanoma cells were highly efficient at colonising the perivascular niche relative to PTEN-expressing counterparts. PTEN re-expression (PTEN-RE) in melanoma cells significantly reduced brain colonisation and migration along the vasculature. We hypothesised this phenotype was mediated through vascular-induced TGFβ secretion, which drives AKT phosphorylation. Disabling TGFβ signalling in melanoma cells reduced colonisation and perivascular invasion; however, the introduction of constitutively active myristolated-AKT (myrAKT) restored overall tumour size but not perivascular invasion.

Conclusions

PTEN loss facilitates perivascular brain colonisation and invasion of melanoma. TGFβ-AKT signalling partially contributes to this phenotype, but further studies are needed to determine the complementary mechanisms that enable melanoma cells to both survive and spread along the brain vasculature.

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Fig. 1: Melanoma cells deficient in PTEN invade along blood vessels in the brain microenvironment.
Fig. 2: Perivascular invasive melanoma upregulates transcriptional programmes important for adhesion, migration, and survival.
Fig. 3: Pten re-expression reduces perivascular invasion along the brain vasculature in vivo.
Fig. 4: Pten re-expression reduces melanoma adhesion to brECs and inhibits migration on ex vivo brain slices.
Fig. 5: TGFβ is upregulated in brEC:melanoma co-cultures.
Fig. 6: TGFβ signalling contributes to brain colonisation and perivascular invasion.

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Data availability

Bioinformatics data will be made available immediately upon publication. Data can be accessed from the NCBI GEO database under accession code GSE247232 or the NCBI SRA database under the accession code PRJNA1032415.

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Acknowledgements

We would like to thank the UVA Flow Cytometry Core, particularly Alex Wendling and Taylor Harper, for their assistance with Luminex and FACS experiments. The UVA Bioinformatics Core, including Pankaj Kumar, assisted with bulk RNA sequencing analysis. The UVA Biospecimen and Tissue Research Facility provided use of equipment for cryosections (Leica microtome). We would also like to acknowledge Drs Kimberly Stegmaier and Brian Crompton, who provided the CRISPR plasmids and protocols. Sonia Patel and Yuvraj Sethi assisted with genotyping mice. BioRender was used to design all schematics.

Funding

SW is supported by the National Institute of Health/National Cancer Institute (NIH/NCI) Ruth L. Kirschstein NRSA for Individual Predoctoral Fellows Award (5F30CA268842-02) and previously received support from the NIH/NCI T32 CA009109 and NIH/NIGMS T32 GM007267 training grants, a trainee award from the UVA Comprehensive Cancer Center, and a medical student research award from the Melanoma Research Foundation. CPR is supported by the NIH/NCI training grant T32 CA009109. ACD is supported by grants from the National Institutes of Health/National Cancer Institute (2RO1 CA177875 and RO1 CA2558451), the Melanoma Research Alliance (ID612638), and funds from the Emily Couric Cancer Center at the University of Virginia. Portions of this research were supported by the NCI Cancer Center Support Grant 5P30CA044579 and by the UVA Genome Analysis and Technology Core (RRID:SCR_018883). Additional support was provided by the University of Virginia Flow Cytometry Core (RRID:SCR_018883). While the authors were supported by the NIH, the content of this publication is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.

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Authors and Affiliations

  1. Department of Microbiology, Immunology, and Cancer Biology, The University of Virginia, Charlottesville, VA, 22908, USA

    Sarah Wang, Caroline P. Riedstra, Yu Zhang, Swetha Anandh & Andrew C. Dudley

  2. The University of Virginia Comprehensive Cancer Center, Charlottesville, VA, USA

    Andrew C. Dudley

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Contributions

SW and ACD conceptualised the study and wrote the manuscript. SW carried out the experiments. CPR and YZ assisted with in vivo and in vitro functional studies. SA assisted with additional experiments.

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Correspondence to Andrew C. Dudley.

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Wang, S., Riedstra, C.P., Zhang, Y. et al. PTEN-restoration abrogates brain colonisation and perivascular niche invasion by melanoma cells. Br J Cancer 130, 555–567 (2024). https://doi.org/10.1038/s41416-023-02530-5

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