Abstract
BackgroundGlioblastomas stem-like cells (GSCs) by invading the brain parenchyma, remains after resection and radiotherapy and the tumoral microenvironment become stiffer. GSC invasion is reported as stiffness sensitive and associated with altered N-glycosylation pattern. Glycocalyx thickness modulates integrins mechanosensing, but details remain elusive and glycosylation enzymes involved are unknown. Here, we studied the association between matrix stiffness modulation, GSC migration and MGAT5 induced N-glycosylation in fibrillar 3D context.MethodTo mimic the extracellular matrix fibrillar microenvironments, we designed 3D-ex-polyacrylonitrile nanofibers scaffolds (NFS) with adjustable stiffnesses by loading multiwall carbon nanotubes (MWCNT). GSCs neurosphere were plated on NFSs, allowing GSCs migration and MGAT5 was deleted using CRISPR-Cas9.ResultsWe found that migration of GSCs was maximum at 166 kPa. Migration rate was correlated with cell shape, expression and maturation of focal adhesion (FA), Epithelial to Mesenchymal Transition (EMT) proteins and (β1,6) branched N-glycan binding, galectin-3. Mutation of MGAT5 in GSC inhibited N-glycans (β1–6) branching, suppressed the stiffness dependence of migration on 166 kPa NFS as well as the associated FA and EMT protein expression.ConclusionMGAT5 catalysing multibranched N-glycans is a critical regulators of stiffness induced invasion and GSCs mechanotransduction, underpinning MGAT5 as a serious target to treat cancer.
Highlights
Glioblastomas stem-like cells (GSCs) by invading the brain parenchyma, remains after resection and radiotherapy and the tumoral microenvironment become stiffer
We initially developed a nanofibers scaffolds (NFS) made of 3D electrospun ex-polyacrylonitrile which supported GSC migration in vitro reflecting the behaviour of glioblastomas in vivo [7]
In this paper we report the mechanosensing interaction existing between substratum stiffness and MGAT5 activity by profiling wild-type GSCs (WT GSC) and MGAT5 knock-out GSCs (MGAT5 KO GSC) seeded onto NFSs made with stiffnesses from 3 to 1260 kPa
Summary
Glioblastomas stem-like cells (GSCs) by invading the brain parenchyma, remains after resection and radiotherapy and the tumoral microenvironment become stiffer. We studied the association between matrix stiffness modulation, GSC migration and MGAT5 induced N-glycosylation in fibrillar 3D context. Extra Cellular Matrix (ECM) stiffness is reported to increase during tumor growth and from low grade glioma to GBM [3], whereby tissue stiffness has been suggested to modulate GSCs migration capacity [2] and tumour progression. These gliomainduced changes in ECM stiffness are accompanied by changes of the glioma cell morphology and nuclear volume [4] and promote epithelial to mesenchymal transition (EMT) [2]. Little is known about stiffness impact of fibrillar 3D environment on migration
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