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Shift of microRNA profile upon orthotopic xenografting of glioblastoma spheroid cultures

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Abstract

Glioblastomas always recur despite surgery, radiotherapy and chemotherapy. A key player in the therapeutic resistance may be immature tumor cells with stem-like properties (TSCs) escaping conventional treatment. A group of promising molecular targets are microRNAs (miRs). miRs are small non-coding RNAs exerting post-transcriptional regulation of gene expression. In this study we aimed to identify over-expressed TSC-related miRs potentially amenable for therapeutic targeting. We used non-differentiated glioblastoma spheroid cultures (GSCs) containing TSCs and compared these to xenografts using a NanoString nCounter platform. This revealed 19 over-expressed miRs in the non-differentiated GSCs. Additionally, non-differentiated GSCs were compared to neural stem cells (NSCs) using a microarray platform. This revealed four significantly over-expressed miRs in the non-differentiated GSCs in comparison to the NSCs. The three most over-expressed miRs in the non-differentiated GSCs compared to xenografts were miR-126, -137 and -128. KEGG pathway analysis suggested the main biological function of these over-expressed miRs to be cell-cycle arrest and diminished proliferation. To functionally validate the profiling results suggesting association of these miRs with stem-like properties, experimental over-expression of miR-128 was performed. A consecutive limiting dilution assay confirmed a significantly elevated spheroid formation in the miR-128 over-expressing cells. This may provide potential therapeutic targets for anti-miRs to identify novel treatment options for GBM patients.

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Acknowledgments

This work was supported by Grants from Odense University Hospital, Region of Southern Denmark, Familien Erichsens Foundation, Svend Helge Arvid Schrøder og Hustru Ketty Larsen Foundation, by the Lundbeckfonden Grant for the NanoCAN Center of Excellence in Nanomedicine, the DAWN-2020 Project financed by Rektorspuljen SDU2020 program, and the MIO Project of the OUH Frontlinjepuljen. We are grateful for the excellent technical assistance from Helle Wohlleben and Tanja D. Højgaard, Department of Pathology, Odense University Hospital.

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Authors and Affiliations

  1. Department of Pathology, Odense University Hospital, Winsløwparken 15, 5000, Odense C, Denmark

    Bo Halle, Sune Munthe, Mia D. Sørensen, Stine S. Jensen & Bjarne W. Kristensen

  2. Department of Neurosurgery, Odense University Hospital, Sdr. Boulevard 29, 5000, Odense C, Denmark

    Bo Halle, Sune Munthe, Mette K. Schulz & Claus Andersen

  3. Institute of Clinical Research, University of Southern Denmark, Winsløwparken 19.3, 5000, Odense C, Denmark

    Bo Halle, Mads Thomassen, Sune Munthe, Mia D. Sørensen, Stine S. Jensen, Torben A. Kruse, Mette K. Schulz, Claus Andersen & Bjarne W. Kristensen

  4. Department of Clinical Genetics, Odense University Hospital, Sdr. Boulevard 29, 5000, Odense C, Denmark

    Mads Thomassen & Torben A. Kruse

  5. Regulus Therapeutics, 3545 John Hopkins Ct., Suite 210, San Diego, CA, 92121-1121, USA

    Ranga Venkatesan, Vivek Kaimal & Eric G. Marcusson

  6. Department of Nuclear Medicine, Odense University Hospital, Sdr. Boulevard 29, 5000, Odense C, Denmark

    Charlotte Aaberg-Jessen

  7. Department of Neurobiology Research, Institute of Molecular Medicine, University of Southern Denmark, Winsløwparken 21, 5000, Odense C, Denmark

    Morten Meyer

  8. Faculty of Health Sciences, Lundbeckfonden Center of Excellence NanoCAN and Molecular Oncology, Institute of Molecular Medicine, University of Southern Denmark, Winsløwparken 25, 5000, Odense C, Denmark

    Helle Christiansen, Steffen Schmidt & Jan Mollenhauer

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  1. Bo Halle
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Correspondence to Bo Halle.

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The experiments in this study complied with the laws of the countries in which they were performed.

Conflict of interest

Eric Marcusson was Senior Director of Oncology, Ranga Venkatesan was Senior Scientist and Vivek Kaimal is Scientist, Bioinformatics at Regulus Therapeutics. All other authors declare no conflict of interest.

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Halle, B., Thomassen, M., Venkatesan, R. et al. Shift of microRNA profile upon orthotopic xenografting of glioblastoma spheroid cultures. J Neurooncol 128, 395–404 (2016). https://doi.org/10.1007/s11060-016-2125-x

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