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Treatment with 5-azacitidine delay growth of glioblastoma xenografts: a potential new treatment approach for glioblastomas

  • Original Article – Cancer Research
  • Published:
Journal of Cancer Research and Clinical Oncology Aims and scope Submit manuscript

Abstract

Purpose

Glioblastoma multiforme (GBM) is the most lethal primary brain tumor in adults. The epigenetically active ribonucleoside analog 5-azacitidine is a new therapy option that changes tumor cell chromatin, which is frequently modified by methylation and deacetylation in malignant gliomas.

Methods

In vitro, we analyzed cell viability, cell apoptosis, and migration of human GBM cells. In vivo, we established subcutaneous and intracerebral GBM mouse models originating from U87MG, U373MG, and primary GBM cells as well as one patient-derived xenograft. Xenografts were treated with 5-azacitidine as well as valproic acid, bevacizumab, temozolomide, and phosphate buffered saline. The tumor sizes and Ki67 proliferation indices were determined. Glioma angiogenesis was examined immunohistochemically by expression analysis of endothelial cells (CD31) and pericytes (PDGFRβ).

Results

In vitro, 5-azacitidine treatment significantly reduced human glioblastoma cell viability, increased cellular apoptosis, and reduced cellular migration. In vivo, 5-azacitidine significantly reduced growth in two intracerebral GBM models. Notably, this was also shown for a xenograft established from a patient surgery sample; whereas, epigenetically acting valproic acid did not show any growth reduction. Highly vascularized tumors responded to treatment, whereas low-vascularized xenografts showed no response. Furthermore, intracerebral glioblastomas treated with 5-azacitidine showed a clearly visible reduction of tumor angiogenesis and lower numbers of endothelial cells and tumor vessel pericytes.

Conclusions

Our data show significant growth inhibition as well as antiangiogenic effects in intracerebral as well as patient-derived GBM xenografts. This encourages to investigate in detail the multifactorial effects of 5-azacitidine on glioblastomas.

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Acknowledgements

We would like to thank Dr. J. Walter (Department of Neurosurgery; University Hospital of Jena; Jena, Germany) for providing human primary glioblastoma cells. We would also like to thank Margit Lemm and Carsta Werner (MaxDelbrückCenter for Molecular Medicine; Berlin, Germany) for excellent technical assistance.

Funding

No funding received.

Author information

Authors and Affiliations

  1. Department of Neurosurgery, Charité University Hospital, Chariteplatz 1, 10117, Berlin, Germany

    Tobias Kratzsch & Peter Vajkoczy

  2. Department of Neurosurgery, Ernst von Bergmann Hospital, Potsdam, Germany

    Susanne Antje Kuhn

  3. Department of Neurosurgery, Vivantes Hospital Berlin Neukölln, Berlin, Germany

    Andreas Joedicke

  4. Institute of Neuropathology, University Hospital, Göttingen, Germany

    Uwe Karsten Hanisch

  5. Paul Flechsig Institute for Brain Research, University of Leipzig, Leipzig, Germany

    Uwe Karsten Hanisch

  6. Experimental Pharmacology and Oncology GmbH, Berlin, Germany

    Jens Hoffmann

  7. Max Delbrueck Center for Molecular Medicine, Berlin, Germany

    Iduna Fichtner

Authors
  1. Tobias Kratzsch
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  2. Susanne Antje Kuhn
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  3. Andreas Joedicke
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  4. Uwe Karsten Hanisch
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  5. Peter Vajkoczy
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  6. Jens Hoffmann
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  7. Iduna Fichtner
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Corresponding author

Correspondence to Tobias Kratzsch.

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Conflict of interest

All authors declare that they have no conflicts of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Informed consent was obtained from all individual participants included in the study.

Additional information

This study is dedicated to Prof. Uwe-Karsten Hanisch.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Suppl. 1

Five-azacitidine elicited no antiangiogenic effects in scanty vascularized CX1 and CX2 GBM xenografts. (A, B) In CX1 GBMs, CD31 labeling revealed no significant reduction of the number of endothelial cells after 5-azacitidine therapy (p > 0.05), in comparison to the antiangiogenic effects of the positive controls bevacizumab and temozolomide (p < 0.001). (C, D) Vascularization was also not significantly affected by 5-azacitidine therapy in CX2 xenografts (p > 0.05), compared to the reduced number in the positive controls bevacizumab (p < 0.001) and temozolomide (p < 0.05), as demonstrated by CD31 labeling. (E) Otherwise, in highly vascularized subcutaneous U87MG GBMs, 5-azacitidine as well as bevacizumab therapy significantly (p < 0.001) reduced the number of CD31 positive endothelial cells. (PPT 651 KB)

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Kratzsch, T., Kuhn, S.A., Joedicke, A. et al. Treatment with 5-azacitidine delay growth of glioblastoma xenografts: a potential new treatment approach for glioblastomas. J Cancer Res Clin Oncol 144, 809–819 (2018). https://doi.org/10.1007/s00432-018-2600-1

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  • DOI: https://doi.org/10.1007/s00432-018-2600-1

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