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Updated: Aug 4, 2025
Characterization of Immune Cell-derived Extracellular Vesicles and Studying Functional Impact on Cell Environment
Published on: June 2, 2020
Small extracellular vesicles promote invadopodia activity in glioblastoma cells in a therapy-dependent manner
Clarissa A Whitehead1, Haoyun Fang2, Huaqi Su3,4
1Department of Surgery, Royal Melbourne Hospital, The University of Melbourne, Parkville, VIC, Australia.
Purpose:
The therapeutic efficacy of radiotherapy/temozolomide treatment for glioblastoma (GBM) is limited by the augmented invasiveness mediated by invadopodia activity of surviving GBM cells. As yet, however the underlying mechanisms remain poorly understood. Due to their ability to transport oncogenic material between cells, small extracellular vesicles (sEVs) have emerged as key mediators of tumour progression. We hypothesize that the sustained growth and invasion of cancer cells depends on bidirectional sEV-mediated cell-cell communication.
Methods:
Invadopodia assays and zymography gels were used to examine the invadopodia activity capacity of GBM cells. Differential ultracentrifugation was utilized to isolate sEVs from conditioned medium and proteomic analyses were conducted on both GBM cell lines and their sEVs to determine the cargo present within the sEVs. In addition, the impact of radiotherapy and temozolomide treatment of GBM cells was studied.
Results:
We found that GBM cells form active invadopodia and secrete sEVs containing the matrix metalloproteinase MMP-2. Subsequent proteomic studies revealed the presence of an invadopodia-related protein sEV cargo and that sEVs from highly invadopodia active GBM cells (LN229) increase invadopodia activity in sEV recipient GBM cells. We also found that GBM cells displayed increases in invadopodia activity and sEV secretion post radiation/temozolomide treatment. Together, these data reveal a relationship between invadopodia and sEV composition/secretion/uptake in promoting the invasiveness of GBM cells.
Conclusions:
Our data indicate that sEVs secreted by GBM cells can facilitate tumour invasion by promoting invadopodia activity in recipient cells, which may be enhanced by treatment with radio-chemotherapy. The transfer of pro-invasive cargos may yield important insights into the functional capacity of sEVs in invadopodia.
Insights
Glioblastoma cells use small extracellular vesicles (sEVs) to enhance invasion by promoting invadopodia activity. This process may be amplified by radiotherapy/temozolomide treatment, impacting glioblastoma progression.
Area of Science:
- Oncology
- Cell Biology
- Biochemistry
Background:
- Glioblastoma (GBM) treatment efficacy is limited by invasive surviving cells.
- Small extracellular vesicles (sEVs) mediate tumor progression by transferring oncogenic material.
- Mechanisms of GBM invasiveness and treatment resistance are poorly understood.
Purpose of the Study:
- To investigate the role of sEVs in GBM cell invasion.
- To explore the relationship between invadopodia activity and sEVs in GBM.
- To determine if radiotherapy/temozolomide treatment affects sEV-mediated invasion.
Main Methods:
- Invadopodia assays and zymography to assess GBM cell invasiveness.
- Differential ultracentrifugation to isolate sEVs.
- Proteomic analysis of GBM cells and their sEVs to identify cargo.
- Assessment of GBM cells post-radiotherapy/temozolomide treatment.
Main Results:
- GBM cells secrete sEVs containing MMP-2 and invadopodia-related proteins.
- sEVs from highly invasive GBM cells enhance invadopodia activity in recipient cells.
- Radiotherapy/temozolomide treatment increases GBM cell invadopodia activity and sEV secretion.
Conclusions:
- GBM-secreted sEVs promote tumor invasion by enhancing recipient cell invadopodia activity.
- Radio-chemotherapy may enhance this sEV-mediated invasion.
- sEV cargo transfer offers insights into GBM invasiveness and therapeutic resistance.
