Brain and Breast Cancer Cells with PTEN Loss of Function Reveal Enhanced Durotaxis and RHOB Dependent Amoeboid Migration Utilizing 3D Scaffolds and Aligned Microfiber Tracts
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Annalena Wieland1, Pamela L Strissel1,2,3, Hannah Schorle1
1Laboratory for Molecular Medicine, Comprehensive Cancer Center Erlangen-EMN (CCC ER-EMN), Department of Gynecology and Obstetrics, University Hospital Erlangen, Friedrich-Alexander University Erlangen-Nuernberg, Universitaetsstrasse 21-23, 91054 Erlangen, Germany.
Background:
Glioblastoma multiforme (GBM) and metastatic triple-negative breast cancer (TNBC) with PTEN mutations often lead to brain dissemination with poor patient outcome, thus new therapeutic targets are needed. To understand signaling, controlling the dynamics and mechanics of brain tumor cell migration, we implemented GBM and TNBC cell lines and designed 3D aligned microfibers and scaffolds mimicking brain structures.
Methods:
3D microfibers and scaffolds were printed using melt electrowriting. GBM and TNBC cell lines with opposing PTEN genotypes were analyzed with RHO-ROCK-PTEN inhibitors and PTEN rescue using live-cell imaging. RNA-sequencing and qPCR of tumor cells in 3D with microfibers were performed, while scanning electron microscopy and confocal microscopy addressed cell morphology.
Results:
In contrast to the PTEN wildtype, GBM and TNBC cells with PTEN loss of function yielded enhanced durotaxis, topotaxis, adhesion, amoeboid migration on 3D microfibers and significant high RHOB expression. Functional studies concerning RHOB-ROCK-PTEN signaling confirmed the essential role for the above cellular processes.
Conclusions:
This study demonstrates a significant role of the PTEN genotype and RHOB expression for durotaxis, adhesion and migration dependent on 3D. GBM and TNBC cells with PTEN loss of function have an affinity for stiff brain structures promoting metastasis. 3D microfibers represent an important tool to model brain metastasizing tumor cells, where RHO-inhibitors could play an essential role for improved therapy.
Insights
Loss of PTEN in glioblastoma and breast cancer cells enhances brain metastasis. RHOB signaling drives this migration, suggesting RHO-inhibitors as potential therapies for brain tumors.
Area of Science:
- Oncology
- Biophysics
- Cell Biology
Background:
- Glioblastoma multiforme (GBM) and triple-negative breast cancer (TNBC) with PTEN mutations are linked to poor outcomes and brain metastasis.
- New therapeutic targets are crucial for managing brain dissemination in these aggressive cancers.
Purpose of the Study:
- To investigate the signaling pathways controlling brain tumor cell migration dynamics and mechanics.
- To model brain tumor cell migration using 3D microfibers and scaffolds that mimic brain tissue.
Main Methods:
- Utilized melt electrowriting to create 3D microfibers and scaffolds.
- Analyzed GBM and TNBC cell lines with varying PTEN genotypes using RHO-ROCK-PTEN inhibitors and PTEN rescue.
- Employed live-cell imaging, RNA-sequencing, qPCR, scanning electron microscopy, and confocal microscopy.
Main Results:
- PTEN-deficient GBM and TNBC cells exhibited increased durotaxis, topotaxis, adhesion, and amoeboid migration on 3D microfibers.
- These cells showed significantly higher RHOB expression compared to PTEN wildtype cells.
- RHOB-ROCK-PTEN signaling was confirmed to be essential for these migratory processes.
Conclusions:
- PTEN genotype and RHOB expression critically influence 3D-dependent cell migration, adhesion, and durotaxis.
- PTEN-deficient GBM and TNBC cells show an affinity for stiff brain structures, promoting metastasis.
- 3D microfibers are valuable tools for modeling brain-metastasizing tumor cells, highlighting the therapeutic potential of RHO-inhibitors.
