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Isolation of Cells from Glioblastoma Multiforme Grade 4 Tumors for Infection with Zika Virus prME and ME Pseudotyped
Celine Pöhlking1, Sebastian Beier1, Jan Patrick Formanski1
1Department of Virology, LG-Schreiber, Bernhard Nocht Institute for Tropical Medicine, Bernhard Nocht Str. 74, 20359 Hamburg, Germany.
Abstract:
This study aimed to isolate cells from grade 4 glioblastoma multiforme tumors for infection experiments with Zika virus (ZIKV) prME or ME enveloped HIV-1 pseudotypes. The cells obtained from tumor tissue were successfully cultured in human cerebrospinal fluid (hCSF) or a mixture of hCSF/DMEM in cell culture flasks with polar and hydrophilic surfaces. The isolated tumor cells as well as the U87, U138, and U343 cells tested positive for ZIKV receptors Axl and Integrin αvβ5. Pseudotype entry was detected by the expression of firefly luciferase or green fluorescent protein (gfp). In prME and ME pseudotype infections, luciferase expression in U-cell lines was 2.5 to 3.5 logarithms above the background, but still two logarithms lower than in the VSV-G pseudotype control. Infection of single cells was successfully detected in U-cell lines and isolated tumor cells by gfp detection. Even though prME and ME pseudotypes had low infection rates, pseudotypes with ZIKV envelopes are promising candidates for the treatment of glioblastoma.
Insights
Researchers explored using Zika virus (ZIKV) envelopes on HIV pseudotypes to infect glioblastoma cells. These ZIKV-enveloped pseudotypes show potential for glioblastoma treatment, despite initial low infection rates.
Area of Science:
- Neuroscience
- Virology
- Oncology
Background:
- Glioblastoma multiforme (GBM) is an aggressive brain tumor with limited treatment options.
- Zika virus (ZIKV) infects cells by binding to specific receptors, including Axl and Integrin αvβ5.
- Gene therapy and targeted viral delivery are potential therapeutic strategies for GBM.
Purpose of the Study:
- To isolate and culture primary glioblastoma cells for infection experiments.
- To evaluate the infectivity of Zika virus (ZIKV) prME or ME enveloped HIV-1 pseudotypes in glioblastoma cells.
- To assess the potential of ZIKV-enveloped pseudotypes as a therapeutic delivery system for glioblastoma.
Main Methods:
- Isolation and culture of grade 4 glioblastoma cells using human cerebrospinal fluid (hCSF).
- Testing for ZIKV receptors (Axl, Integrin αvβ5) on primary glioblastoma cells and U87, U138, U343 cell lines.
- Infection assays using ZIKV prME/ME enveloped HIV-1 pseudotypes, detecting entry via luciferase or GFP expression.
Main Results:
- Primary glioblastoma cells were successfully cultured and expressed ZIKV receptors.
- ZIKV pseudotypes demonstrated measurable entry into glioblastoma cells, detected by reporter gene expression.
- Infection rates with ZIKV pseudotypes were lower than control VSV-G pseudotypes but above background levels.
Conclusions:
- Zika virus receptors are present on glioblastoma cells, suggesting susceptibility to ZIKV-based vectors.
- ZIKV-enveloped pseudotypes can infect glioblastoma cells, indicating potential for targeted delivery.
- Despite low initial efficiency, ZIKV envelopes show promise for developing novel glioblastoma therapies.
