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Investigation of the tumoricidal effects of sonodynamic therapy in malignant glioblastoma brain tumors

Abstract

Objective

Glioblastoma is the most common primary brain tumor; survival is typically 12–18 months after diagnosis. We sought to study the effects of sonodynamic therapy (SDT) using 5-Aminolevulinic acid hydrochloride (5-ALA) and high frequency focused ultrasound (FUS) on 2 glioblastoma cell lines.

Procedure

Rat C6 and human U87 glioblastoma cells were studied under the following conditions: 1 mM 5-ALA (5-ALA); focused ultrasound (FUS); 5-ALA and focused ultrasound (SDT); control. Studied responses included cell viability using an MTT assay, microscopic changes using phase contract microscopy, apoptotic induction through a caspase-3 assay, and apoptosis staining to quantify cell death.

Results

SDT led to a marked decrease in cell extension and reduction in cell size. For C6, the MTT assay showed reductions in cell viability for 5-ALA, FUS, and SDT groups of 5%, 16%, and 47%, respectively compared to control (p < 0.05). Caspase 3 induction in C6 cells relative to control showed increases of 109%, 110%, and 278% for 5-ALA, FUS, and SDT groups, respectively (p < 0.05). For the C6 cells, caspase 3 staining positivity was 2.1%, 6.7%, 11.2%, and 39.8% for control, 5-ALA, FUS, and SDT groups, respectively. C6 Parp-1 staining positivity was 1.9%, 6.5%, 9.0%, and 37.8% for control, 5-ALA, FUS, and SDT groups, respectively. U87 cells showed similar responses to the treatments.

Conclusions

Sonodynamic therapy resulted in appreciable glioblastoma cell death as compared to 5-ALA or FUS alone. The approach couples two already FDA approved techniques in a novel way to treat the most aggressive and malignant of brain tumors. Further study of this promising technique is planned.

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Acknowledgements

We are grateful for the support of Drs. Eames and Moore who allowed us to use their 3D CAD software and 3D printer at the Focused Ultrasound Foundation. Finally, we appreciate the assistance of Dr. Pramoonjago at the University of Virginia’s Biorepository and Tissue Research Facility who processed the cells using automated instrumentation for the cytospin techniques for immunohistochemical staining.

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Correspondence to Kimball Sheehan.

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The current research does not involve human participants and/or animals and therefore informed consent was not required.

Disclosures

Dr. Padilla and Dr. Moore are employees of the Focused Ultrasound Foundation. Otherwise, the authors have no disclosures related to this particular study.

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11060_2020_3504_MOESM1_ESM.jpg

Supplemental Figure 1: One of the 3-D printed devices for treating the glioblastoma cells with SDT. The ultrasound probe was slid into the cylindrical column and the plate or dish was placed on the upper flat surface. The entire system was placed in degassed water at the time of treatment. (JPEG 1,907 kb)

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Sheehan, K., Sheehan, D., Sulaiman, M. et al. Investigation of the tumoricidal effects of sonodynamic therapy in malignant glioblastoma brain tumors. J Neurooncol 148, 9–16 (2020). https://doi.org/10.1007/s11060-020-03504-w

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Keywords

  • Glioblastoma
  • Focused ultrasound
  • Sonodynamic therapy
  • Apoptosis