Glioblastoma extracellular vesicles influence glial cell hyaluronic acid deposition to promote invasiveness

Dominik Koessinger; David Novo; Anna Koessinger; America Campos; Jasmine Peters; Louise Dutton; Peggy Paschke; Désirée Zerbst; Madeleine Moore; Louise Mitchell; Matthew Neilson; Katrina Stevenson; Anthony Chalmers; Stephen Tait; Joanna Birch; Jim Norman

doi:10.1093/noajnl/vdad067

Glioblastoma extracellular vesicles influence glial cell hyaluronic acid deposition to promote invasiveness

Neurooncol Adv. 2023 May 27;5(1):vdad067. doi: 10.1093/noajnl/vdad067. eCollection 2023 Jan-Dec.

Authors

Dominik Koessinger^{1

2

3}, David Novo¹, Anna Koessinger^{1

2}, America Campos¹, Jasmine Peters¹, Louise Dutton², Peggy Paschke¹, Désirée Zerbst^{1

2}, Madeleine Moore¹, Louise Mitchell¹, Matthew Neilson¹, Katrina Stevenson², Anthony Chalmers², Stephen Tait^{1

2}, Joanna Birch², Jim Norman^{1

2}

Affiliations

¹ Cancer Research UK Beatson Institute, Glasgow, UK.
² School of Cancer Sciences, University of Glasgow, Glasgow, UK.
³ Department of Neurosurgery, Freiburg University Hospital, Freiburg, Germany.

Abstract

Background: Infiltration of glioblastoma (GBM) throughout the brain leads to its inevitable recurrence following standard-of-care treatments, such as surgical resection, chemo-, and radiotherapy. A deeper understanding of the mechanisms invoked by GBM to infiltrate the brain is needed to develop approaches to contain the disease and reduce recurrence. The aim of this study was to discover mechanisms through which extracellular vesicles (EVs) released by GBM influence the brain microenvironment to facilitate infiltration, and to determine how altered extracellular matrix (ECM) deposition by glial cells might contribute to this.

Methods: CRISPR was used to delete genes, previously established to drive carcinoma invasiveness and EV production, from patient-derived primary and GBM cell lines. We purified and characterized EVs released by these cells, assessed their capacity to foster pro-migratory microenvironments in mouse brain slices, and evaluated the contribution made by astrocyte-derived ECM to this. Finally, we determined how CRISPR-mediated deletion of genes, which we had found to control EV-mediated communication between GBM cells and astrocytes, influenced GBM infiltration when orthotopically injected into CD1-nude mice.

Results: GBM cells expressing a p53 mutant (p53^R273H) with established pro-invasive gain-of-function release EVs containing a sialomucin, podocalyxin (PODXL), which encourages astrocytes to deposit ECM with increased levels of hyaluronic acid (HA). This HA-rich ECM, in turn, promotes migration of GBM cells. Consistently, CRISPR-mediated deletion of PODXL opposes infiltration of GBM in vivo.

Conclusions: This work describes several key components of an EV-mediated mechanism though which GBM cells educate astrocytes to support infiltration of the surrounding healthy brain tissue.

Keywords: GBM; astrocytes; extracellular matrix; extracellular vesicles; invasion.

Abstract

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