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Targeting glioblastoma multiforme using a novel fusion protein comprising interleukin-13 and staphylococcal
Zahra Gholipour1, Abbas Ali Imani Fooladi2, Kazem Parivar1
1Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Abstract:
Targeting cell surface receptors with immunotoxins provides a novel, unique and highly potent treatment against cancers. A high expression of interleukin-13 (IL13) receptor α2 (IL13Rα2) has been reported in different types of cancers including glioblastoma multiforme (GBM). In this paper, to target IL13Rα2 on GBM cells, a fusion protein was generated comprising human IL13 and staphylococcal enterotoxin B (SEB), termed IL13-linker-SEB. The fusion protein was cloned into pET28a(+) and expressed in Escherichia coli strain BL21 (DE3); U251 (IL13Rα2-positive) and T98G (IL13Rα2-negative) GBM cell lines were employed and the functional activity of IL13-linker-SEB was evaluated by cell ELISA, cytotoxicity (MTT and LDH), apoptosis (flow cytometry and caspase-3 activity), adhesion, scratch and RT-PCR tests. SEB and chemotherapeutic drugs were employed to be compared to IL13-linker-SEB function. The IL13-linker-SEB exhibited higher binding affinity and cytotoxicity compared to SEB on U251 cells, although both recombinant proteins had shown similar behavior regarding T98G cells. Furthermore, the highest induction of apoptosis was observed in U251 cells treated with IL13-linker-SEB which was confirmed by Bax/Bcl-2 ratio. The expression of MMP2, MMP9 and VEGFR2 in U251 cells experienced a significant reduction after treatment with IL13-linker-SEB compared to SEB and T98G treated cells. The data showed that IL13-linker-SEB can be considered as a novel potential agent for GBM treatment; however, further research is needed to investigate the efficacy.
Insights
A novel immunotoxin, IL13-linker-SEB, shows potent anti-cancer activity against glioblastoma multiforme (GBM) by targeting the IL13 receptor α2 (IL13Rα2). This fusion protein effectively reduces tumor cell viability and induces apoptosis, offering a promising new therapeutic strategy for GBM.
Area of Science:
- Biotechnology
- Oncology
- Molecular Biology
Background:
- Immunotoxins offer a potent cancer treatment strategy by targeting cell surface receptors.
- Interleukin-13 receptor α2 (IL13Rα2) is highly expressed in glioblastoma multiforme (GBM) and other cancers.
Purpose of the Study:
- To develop and evaluate a novel fusion protein, IL13-linker-SEB, targeting IL13Rα2 for GBM treatment.
- To assess the efficacy of IL13-linker-SEB in vitro against GBM cell lines.
Main Methods:
- A fusion protein (IL13-linker-SEB) was constructed by combining human IL13 and staphylococcal enterotoxin B (SEB).
- The fusion protein was expressed in E. coli and tested on IL13Rα2-positive (U251) and negative (T98G) GBM cell lines.
- Functional assays included cell ELISA, cytotoxicity (MTT, LDH), apoptosis (flow cytometry, caspase-3), adhesion, scratch, and RT-PCR.
Main Results:
- IL13-linker-SEB demonstrated superior binding affinity and cytotoxicity compared to SEB on U251 cells.
- Treatment with IL13-linker-SEB significantly induced apoptosis in U251 cells, evidenced by the Bax/Bcl-2 ratio.
- The expression of MMP2, MMP9, and VEGFR2 was significantly reduced in U251 cells post-treatment with IL13-linker-SEB.
Conclusions:
- IL13-linker-SEB is a promising novel agent for targeting IL13Rα2-positive GBM.
- Further research is warranted to investigate the clinical efficacy of this immunotoxin for GBM therapy.
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