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Heterogeneity of radiation response in mesenchymal subtype glioblastoma: molecular profiling and reactive oxygen species generation

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Abstract

Background

Radiotherapy-induced tumor death remains critical in the successful first-line management of glioblastoma, whereas resistance to radiation serves as a major factor in disease progression. Mesenchymal shift has been identified as a driver in GBM recurrence, with gene expression associated with enhanced repair of macromolecular damage caused by radiation.

Methods

Using distinct mesenchymal subtype GBM cells lines, radiation response was assessed by clonogenic assay and orthotopic mouse tumor model. RNA-sequencing was performed in the setting of increasing radiation dosing while real-time assessment of ROS generation was achieved by the measurement of hydroxyl spin trap adducts via electron paramagnetic resonance.

Results

Radiation-induced cell death determined by clonogenic assay was significantly different at low dose (4-8 Gy) between the resistant U3035 cells and the sensitive U3020 cells. Similar trends were present in the in vivo NSG mouse model following radiation dosing on post-implantation day 7–10, with the rate of reduction in tumor bioluminescence reversing between the U3020 and U3035 cells after the third dose of radiation. Changes in gene expression following radiation determined by RNA-sequencing indicate both U3035 and U3020 cells demonstrate a shift toward more mesenchymal profiles, with concurrent shift away from pro-neural subtype gene expression in the U3020 cells that appeared to develop resistance to radiation in vivo. Persistence of ROS generated following radiation was greater in U3020 cells shown to be more sensitive to radiation.

Conclusions

Despite the same molecular classification, distinct GBM cell lines can demonstrate differential response to radiation and potential for mesenchymal shift associated with radiation resistance.

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Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Funding

Research reported in this publication was supported by the National Institutes of Health under Award Number P20GM121322 to CPC and AB. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. CPC was also supported by U54GM104942 through the NIH.

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Authors and Affiliations

  1. Department of Neurosurgery, West Virginia University, 1 Medical Center Drive, Suite 4300, Morgantown, WV, 26506-9183, USA

    Christopher P. Cifarelli & Angelica Jacques

  2. Department of Radiation Oncology, West Virginia University, Morgantown, WV, USA

    Christopher P. Cifarelli

  3. Department of Biochemistry, West Virginia University, Morgantown, WV, USA

    Andrey Bobko

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  1. Christopher P. Cifarelli
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CPC, AJ, and AB have made substantial contributions to conception and design, or acquisition of data, or analysis and interpretation of data; have been involved in drafting the manuscript or revising it critically for important intellectual content; and have given final approval of the version to be published.

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Correspondence to Christopher P. Cifarelli.

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Conflict of interest

CPC reports personal fees from Carl Zeiss Meditec, Inc, outside of the submitted work. No other conflicts exists for the other authors.

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No human subjects were involved in this work. Animal use was approved by the WVU IACUC (Protocol No.1602000263.2) and includes adherence to the ethical use of laboratory animals.

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Cifarelli, C.P., Jacques, A. & Bobko, A. Heterogeneity of radiation response in mesenchymal subtype glioblastoma: molecular profiling and reactive oxygen species generation. J Neurooncol 152, 245–255 (2021). https://doi.org/10.1007/s11060-021-03707-9

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  • DOI: https://doi.org/10.1007/s11060-021-03707-9

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