Skip to Main content Skip to Navigation
Theses

Mécanotransduction des cellules souches de glioblastome dans un nouveau modèle de culture tridimensionnel : implication de la MGAT5 dans la perception de l'environnement mécanique

Abstract : Glioblastoma stem cells (CSC) have been reported to be sensitive to the mechanical properties of the surrounding tissue/microenvironment and to use the microenvironment stiffness to enhance invasion.In this context, we developed a 3D artificial fibrillary tissue which can allow in vitro recapitulation of the migration behavior observed in vivo. This 3D matrix is highly plastic which allows for modulation of stiffness, surface chemistry and fibers alignment.On the first hand, we functionalized the fibers with extracellular matrix proteins and then we plated GSCs neurospheres (NS) on the developed 3D artificial tissue. The addition of laminin modulates the migration behaviour from single cell to collective mode.On the second hand we have modified stiffness of the fibers. After five days of GSCs NS migration on 3D electrospun fibers of different stiffnesses, we have seen an increase in migration velocity for an intermediate stiffness of 166kPa in comparison with our softest 3.2 kPa and stiffest stiffnesses (1260 kPa). This maximum migration rate is associated with changes in cell shape, increase of EMT proteins expressions and modifications of focal adhesion proteins regulation.Mannoside acetyl glucosaminyltransferase 5 (MGAT5) overexpression is associated with malignant tumors and it is implicated in the clustering of membrane proteins through lattice formation, focal adhesions, cell migration, invasion, and epithelial-to-mesenchymal transition (EMT), resulting in functional advantages for tumor cells.In light of previous results about of EMT proteins expressions and modifications of focal adhesion proteins regulation, we generated MGAT5 CRISPR Cas9 GSCs and placed it on 3D matrix with different stiffnesses. We observe a decrease in migration velocity at the intermediate stiffness in comparison with GSCs WT associated with a decrease in focal adhesion maturation and EMT. This study demonstrates the implication of glycosylation and more particularly MGAT5-mediated glycosylation on GSCs mechanotransduction.
Complete list of metadatas

Cited literature [677 references]  Display  Hide  Download

https://tel.archives-ouvertes.fr/tel-02056673
Contributor : Abes Star :  Contact
Submitted on : Monday, March 4, 2019 - 5:17:07 PM
Last modification on : Tuesday, May 28, 2019 - 5:00:04 PM
Document(s) archivé(s) le : Wednesday, June 5, 2019 - 4:45:50 PM

File

2018_MARHUENDA_archivage.pdf
Version validated by the jury (STAR)

Identifiers

  • HAL Id : tel-02056673, version 1

Collections

Citation

Emilie Marhuenda. Mécanotransduction des cellules souches de glioblastome dans un nouveau modèle de culture tridimensionnel : implication de la MGAT5 dans la perception de l'environnement mécanique. Médecine humaine et pathologie. Université Montpellier, 2018. Français. ⟨NNT : 2018MONTT091⟩. ⟨tel-02056673⟩

Share

Metrics

Record views

230

Files downloads

2808