A novel 3D nanofibre scaffold conserves the plasticity of glioblastoma stem cell invasion by regulating galectin-3 and integrin-\u03b21 expression

Ali Saleh,Hugues Duffau,Norbert Bakalara,Hassan Boukhaddaoui,Luc Bauchet,Jean- Philippe Hugnot,Jan De Weille,Emilie Marhuenda,David Cornu,Sophie Guelfi,Igor Lima Maldonado,Christine Fabre,Zahra Hassani

doi:10.1038/s41598-019-51108-w

Abstract

Glioblastoma Multiforme (GBM) invasiveness renders complete surgical resection impossible and highly invasive Glioblastoma Initiating Cells (GICs) are responsible for tumour recurrence. Their dissemination occurs along pre-existing fibrillary brain structures comprising the aligned myelinated fibres of the corpus callosum (CC) and the laminin (LN)-rich basal lamina of blood vessels. The extracellular matrix (ECM) of these environments regulates GIC migration, but the underlying mechanisms remain largely unknown. In order to recapitulate the composition and the topographic properties of the cerebral ECM in the migration of GICs, we have set up a new aligned polyacrylonitrile (PAN)-derived nanofiber (NF) scaffold. This system is suitable for drug screening as well as discrimination of the migration potential of different glioblastoma stem cells. Functionalisation with LN increases the spatial anisotropy of migration and modulates its mode from collective to single cell migration. Mechanistically, equally similar to what has been observed for mesenchymal migration of GBM in vivo, is the upregulation of galectin-3 and integrin-β1 in Gli4 cells migrating on our NF scaffold. Downregulation of Calpain-2 in GICs migrating in vivo along the CC and in vitro on LN-coated NF underlines a difference in the turnover of focal adhesion (FA) molecules between single-cell and collective types of migration.

Highlights

Glioblastoma Multiforme (GBM) invasiveness renders complete surgical resection impossible and highly invasive Glioblastoma Initiating Cells (GICs) are responsible for tumour recurrence
GICs use a mesenchymal single cell migration mode to migrate away from the main tumour bulk[13] which is characteristic of disseminating glioma[14]. They may form in vivo multicellular networks or clusters implicated in their invasive capacity and radioresistance[15,16]. To recapitulate these different migration modes and to mimic the topography of the white matter tracts the biochemical composition of the brain extracellular matrix (ECM), we developed new NF scaffolds of aligned and non-aligned of stabilized PAN, which are either partially functionalized with LN (+LN) or not (−LN)
The corpus callosum (CC) is the favourite route to the contralateral hemisphere of glioblastoma cells[17]

Summary

Introduction

Glioblastoma Multiforme (GBM) invasiveness renders complete surgical resection impossible and highly invasive Glioblastoma Initiating Cells (GICs) are responsible for tumour recurrence. GICs use a mesenchymal single cell migration mode to migrate away from the main tumour bulk[13] which is characteristic of disseminating glioma[14] They may form in vivo multicellular networks or clusters implicated in their invasive capacity and radioresistance[15,16]. To recapitulate these different migration modes and to mimic the topography of the white matter tracts the biochemical composition of the brain ECM, we developed new NF scaffolds of aligned (aNF) and non-aligned (naNF) of stabilized PAN, which are either partially functionalized with LN (+LN) or not (−LN). We correlated our results with in vivo xenografts of human GIC into the brain of nude mice

Methods

Results

Conclusion