The invasive proteome of glioblastoma revealed by laser-capture microdissection

Daubon, Thomas; Guyon, Joris; Raymond, Anne-Aurélie; Dartigues, Benjamin; Rudewicz, Justine; Ezzoukhry, Zakaria; Dupuy, Jean-William; Herbert, John; Saltel, Frédéric; Bjerkvig, Rolf; Nikolski, Macha; Bikfalvi, Andréas

doi:10.1093/noajnl/vdz029

Cited by 12 publications

(10 citation statements)

References 24 publications

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“…In particular, we have studied the matricellular, thrombospondin-1 (TSP1), and shown that it involved in tumor cell invasion, through the activation of CD47 in tumor cells 12 . Furthermore, using a proteomic approach, we have discovered unexpected role of PLP1 and DNM1 in GBM invasion 17 . In these studies, 3D invasion assays were successfully used with or without pharmacological blockade of TSP1, PLP1 or DNM1.…”

Section: Discussionmentioning

confidence: 99%

“…Glioblastomas are known to extensively invade the surrounding tissues by different processes (co-option, white matter tract invasion, interstitial invasion) 16 . We have recently described two novel mechanisms of glioblastoma invasion 9,12,17 . In particular, we have studied the matricellular, thrombospondin-1 (TSP1), and shown that it involved in tumor cell invasion, through the activation of CD47 in tumor cells 12 .…”

Section: Discussionmentioning

confidence: 99%

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A 3D Spheroid Model for Glioblastoma

Guyon

Andrique

Pujol

et al. 2020

JoVE

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

A 3D Spheroid Model for Glioblastoma

Guyon

Andrique

Pujol

et al. 2020

JoVE

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“…LCM-proteomics allows spatially precise and technically reproducible exploration of the proteomic landscape of human tissue microenvironments. Previously, LCM-proteomic analysis has been applied to frozen tissues from patient-derived xenograph (PDX) glioblastoma models (46). However, PDX models often lack many of the TME constituents present in the primary tumour, including the native human ECM, vasculature, and non-neoplastic cell populations, and thus, may only recapitulate the cellular features of human tumours.…”

Section: Discussionmentioning

confidence: 99%

Novel Laser Capture Microdissection-Proteomic Analysis Identifies Spatially Distinct Extracellular Matrix Signatures in the Core and Infiltrating Edge of Human Glioblastoma

Pedley

Prescott

Appleton

et al. 2022

Preprint

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Background: Glioblastoma is the most common and aggressive primary brain tumour in adults. Hallmarks of glioblastoma include its intra-tumoural heterogeneity and extensive invasion of the surrounding brain. Glioblastoma is known to remodel the extracellular matrix (ECM) of the brain, resulting in altered mechanical properties and the establishment of a tumour-promoting microenvironment. How changes in the expression and spatial distribution of ECM constituents within glioblastoma contribute to invasion and disease progression is still unclear. Methods: Here we report on a protocol for laser-capture microdissection coupled with mass spectrometry (LCM-proteomics) that allowed a spatially resolved and unbiased analysis of the regional ECM proteome (matrisome) in formalin-fixed and paraffin-embedded (FFPE) samples of human glioblastoma. We investigated five molecularly characterised hemispheric adult glioblastomas where the brain/tumour interface and tumour epicentre were represented in the surgical specimens and snap-frozen tissue was available. LCM-proteomic analysis was validated with immunohistochemistry. Results: LCM-proteomics identified 53 matrisome proteins in FFPE tissue, demonstrating comparable performance with conventional analysis of snap-frozen tissue. The analysis revealed distinct matrisome components in the brain/tumour interface versus the tumour epicentre. Guided by data from LCM-proteomic analysis, immunostaining for tenascin-R confirmed greater staining in the brain/tumour interface, whilst expression of fibronectin was higher in the tumour epicentre. Conclusion: The protocol described in this work allowed for accurate, spatially resolved analysis of ECM in FFPE tissues, with performance comparable to analysis of snap-frozen tissue. While the focus for this work was on the regional ECM composition of glioblastoma, we found that the LCM-proteomics protocol is also applicable to the study of the wider proteome, and represents a valuable tool for investigating tumour/tissue heterogeneity. This protocol opens the possibility to apply LCM-proteomics to retrospective studies with the advantage of accessing clinical history and follow-up information, providing a valuable resource for translational research in glioblastoma.

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“…GSCs might also reside in hypoxic and invasive niches. The latest might bolster their migratory and invasive behavior along the vascular structures ( Darmanis et al, 2017 ; Mathivet et al, 2017 ; Griveau et al, 2018 ; Alieva et al, 2019 ; Daubon et al, 2019 ). Accordingly, GBM cells barely disseminate using intravasation within the bloodstream, unlike most peripheral tumor cells that misuse vessel and lymphatic circulation routes ( Rosińska and Gavard, 2021 ).…”

Section: Introductionmentioning

confidence: 99%

Neuropilin-1 modulates the 3D invasive properties of glioblastoma stem-like cells

Kerhervé

Rosińska²,

Trillet³

et al. 2022

Front. Cell Dev. Biol.

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Glioblastoma multiforme (GBM) is a rare, yet devastating, primary brain tumor in adults. Current treatments remain generally ineffective and GBM almost invariably recurs, resulting in median survival of 15 months. This high malignancy sources notably from the resilience and invasive capabilities of tumor cells. Within GBM, exists a population of self-sustaining transformed cells with stem-like properties (GSCs), which are thought to be responsible for tumor initiation, growth, and invasion, as well as recurrence. In the tumor microenvironment, GSCs might be found in the vicinity of brain endothelial cells, which provide a protective habitat. Likewise, these resistant, quiescent GSCs may accumulate in hypoxic zones, away from the perivascular niche, or travel towards the healthy brain parenchyma, by eminently co-opting neuro-vascular tracks. Herein, we established an ex vivo model to explore GSC invasive behavior. We found that patient-derived cells massively invade the collagen matrix. In addition, we described that the glycoprotein Neuropilin-1 (NRP1) contributes to GSC spreading and invasion. Indeed, both RNA interference-mediated silencing and CRISPR-mediated gene editing deletion of NRP1 strongly impaired the 3D invasive properties of patient-derived GSCs and their close localization to the brain blood vessels. Of note, other typical features of GSCs, such as expansion and self-renewal were maintained. From a mechanistic standpoint, this biological effect might rely on the expression of the β3 subunit integrin cell-extracellular matrix adhesive receptor. Our data, therefore, propose a reliable approach to explore invasive properties of patient glioma cells ex vivo and identify NRP1 as a mediator in this malignant process.

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The invasive proteome of glioblastoma revealed by laser-capture microdissection

Cited by 12 publications

References 24 publications

A 3D Spheroid Model for Glioblastoma

A 3D Spheroid Model for Glioblastoma

Novel Laser Capture Microdissection-Proteomic Analysis Identifies Spatially Distinct Extracellular Matrix Signatures in the Core and Infiltrating Edge of Human Glioblastoma

Neuropilin-1 modulates the 3D invasive properties of glioblastoma stem-like cells

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