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A co-formulation of interferons type I and II enhances temozolomide response in glioblastoma with unmethylated MGMT promoter status

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Abstract

Temozolomide (TMZ) is a chemotherapeutic used for the treatment of glioblastoma. The MGMT repair enzyme (O′-(6)-methyl guanine-DNA-methyltransferase) promoter methylation is a predictive biomarker to TMZ response; interferons (IFNs) type I can downregulate MGMT expression improving survival in patients with unmethylated MGMT promoter. HeberFERON is a co-formulation of IFNs type I and II with higher antiproliferative effect over glioblastoma cell lines than individual IFNs. We investigated the proliferative response of patient-derived glioblastoma cultures to HeberFERON and its combination with TMZ in relation to MGMT promoter methylation and the regulation of MGMT transcript after HeberFERON treatment. Eleven glioblastoma-derived cultures, molecularly classified according to TCGA and MGMT promoter methylation, were assayed for proliferation inhibition with HeberFERON at low doses (1–25 IU/mL) [alone or combined with TMZ] or at higher doses (50–200 IU/mL) using CellTiter-Glo Luminescent Cell Viability Assay (Promega). Eight cultures were further treated with 100 IU/mL of HeberFERON for 72 h, total RNA purified (Qiagen) and converted to cDNA (Superscript III kit, Invitrogen) as quantitative PCR templates. Changes of MGMT&P53 transcripts level were monitored. Response of cultures to HeberFERON is variable, dose-dependent and apparently independent from TCGA classification and MGMT methylation status, based on the eight Classical cultures data. When combining HeberFERON with TMZ there was an increase in cell death for cultures, 2/4 with methylated and 5/5 with unmethylated MGMT promoter. In two out five cultures with unmethylated MGMT status, we observed a decrease of MGMT gene levels and an increase in P53 encoding gene levels. HeberFERON and TMZ combination should be further assayed in glioblastoma, mainly for those with unmethylated MGMT promoter.

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Available antiproliferative and gene expression analysis data will be available through the corresponding author.

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Acknowledgements

We would like to thank to Pharmacogenomic group at CIGB and Neurosurgery Department at ErasmusMC for their technical support. Special thanks to Jan-willem Jachtenberg, formerly from Neurosurgery Department at ErasmusMC, for the initial testing of glioblastoma patient-derived cultures with HeberFERON. Part of the work was carried out as part of a UICC International Cancer Technology Transfer Fellowship (ICR/2014/328167).

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Authors and Affiliations

  1. Pharmacogenomic Group, Biomedical Research Direction, Center for Genetic Engineering and Biotechnology, Havana, Cuba

    Dania Vázquez-Blomquist & Adelaida Villarreal

  2. Clinical Assay Direction, Center for Genetic Engineering and Biotechnology, Havana, Cuba

    Iraldo Bello-Rivero

  3. Neurosurgical Department, Erasmus MC, Rotterdam, The Netherlands

    Sieger Leenstra & Mariëlle van der Kaaij

Authors
  1. Dania Vázquez-Blomquist
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  2. Sieger Leenstra
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  4. Adelaida Villarreal
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  5. Iraldo Bello-Rivero
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Corresponding author

Correspondence to Dania Vázquez-Blomquist.

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Conflict of interest

DVB, AV and IBR are employees of the Center for Genetic Engineering and Biotechnology, in Havana where HeberFERON is produced. The authors have no competing interests.

Ethical approval

The research protocol and patient informed consent form that specified the use of patient material for research on brain tumors neurology, neurosurgery and pathology was approved by the Medical Ethics Review Committee (MERC; METC in Dutch) of ErasmusMC (MEC 221.520/2002/262). The research protocol was conducted under the responsibility of the ErasmusMC departments of Neurology, Neurosurgery and Pathology.

Informed consent

Primary glioblastoma cell cultures were derived from tumors of operated patients from the Erasmus Medical Center Rotterdam, The Netherlands. Prior to surgery, patients sign informed consent. All authors consent for publication of the results included in this article.

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Vázquez-Blomquist, D., Leenstra, S., van der Kaaij, M. et al. A co-formulation of interferons type I and II enhances temozolomide response in glioblastoma with unmethylated MGMT promoter status. Mol Biol Rep 47, 5263–5271 (2020). https://doi.org/10.1007/s11033-020-05604-2

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  • DOI: https://doi.org/10.1007/s11033-020-05604-2

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