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.
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.
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.
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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Omuro A, DeAngelis LM (2013) Glioblastoma and other malignant gliomas: a clinical review. JAMA 310(17):1842–1850
McDannold N, Clement GT, Black P, Jolesz F, Hynynen K (2010) Transcranial magnetic resonance imaging—guided focused ultrasound surgery of brain tumors: initial findings in 3 patients. Neurosurgery 66(2):323–332
Walters H, Shah BB (2019) Focused ultrasound and other lesioning therapies in movement disorders. Curr Neurol Neurosci Rep 19(9):66
Fishman PS, Frenkel V (2017) Focused ultrasound: an emerging therapeutic modality for neurologic disease. Neurotherapeutics 14(2):393–404
Zhu L, Altman MB, Laszlo A et al (2019) Ultrasound hyperthermia technology for radiosensitization. Ultrasound Med Biol 45(5):1025–1043
McHale AP, Callan JF, Nomikou N, Fowley C, Callan B (2016) Sonodynamic therapy: concept, mechanism and application to cancer treatment. Adv Exp Med Biol 880:429–450
Yang Y, Tu J, Yang D, Raymond JL, Roy RA, Zhang D (2019) Photo- and sono-dynamic therapy: a review of mechanisms and considerations for pharmacological agents used in therapy incorporating light and sound. Curr Pharm Des 25(4):401–412
Endo S, Kudo N, Yamaguchi S, Sumiyoshi K, Motegi H, Kobayashi H, Terasaka S, Houkin K (2015) Porphyrin derivatives-mediated sonodynamic therapy for malignant gliomas in vitro. Ultrasound Med Biol 41(9):2458–2465
Jeong EJ, Seo SJ, Ahn YJ, Choi KH, Kim KH, Kim JK (2012) Sonodynamically induced antitumor effects of 5-Aminolevulinic acid and fractionated ultrasound irradiation in an orthotopic rat glioma model. Ultrasound Med Biol 38(12):2143–2150
Lafond M, Yoshizawa S, Umemura SI (2019) Sonodynamic therapy: advances and challenges in clinical translation. J Ultrasound Med 38(3):567–580
Pan X, Wang H, Wang S, Sun X, Wang L, Wang W, Shen H, Liu H (2018) Sonodynamic therapy (SDT): a novel strategy for cancer nanotheranostics. Sci China Lif Sci 61(4):415–429
Hersh DS, Kim AJ, Winkles JA, Eisenberg HM, Woodworth GF, Frenkel V (2016) Emerging applications of therapeutic ultrasound in neuro-oncology: moving beyond tumor ablation. Neurosurgery 79(5):643–654
Hadjipanayis CG, Widhalm G, Stummer W (2015) What is the surgical benefit of utilizing 5-Aminolevulinic acid for fluorescence-guided surgery of malignant gliomas? Neurosurgery 77(5):663–673
Li YJ, Huang P, Jiang CL, de Jia X, Du XX, Zhou JH, Han Y, Sui H, Wei XL, Liu L, Yuan HH, Zhang TT, Zhang WJ, Xie R, Lang XH, Wang LY, Liu T, Bai YX, Tian Y (2014) Sonodynamically induced anti-tumor effect of 5-Aminolevulinic acid on pancreatic cancer cells. Ultrasound Med Biol 40(11):2671–2679
Li Y, Zhou Q, Hu Z, Yang B, Li Q, Wang J, Zheng J, Cao W (2015) 5-Aminolevulinic acid-based sonodynamic therapy induces the apoptosis of osteosarcoma in mice. PLoS ONE 10(7):e0132074
Ji C, Yang B, Yang YL, He SH, Miao DS, He L, Bi ZG (2010) Exogenous cell-permeable C6 ceramide sensitizes multiple cancer cell lines to Doxorubicin-induced apoptosis by promoting AMPK activation and mTORC1 inhibition. Oncogene 29(50):6557–6568
Saraste A, Pulkki K (2000) Morphologic and biochemical hallmark of Apoptosis. Cardiovasc Res 45(3):528–537
D’Amours D, Sallmann FR, Dixit VM, Poirier GG (2001) Gain-of-function of poly(ADP-ribose) polymerase-1 upon cleavage by apoptotic proteases: implications for apoptosis. J Cell Sci 114(20):3771–3778
Grisham J (2015) The Tumor. eBook.
Fomenko A, Lozano AM (2019) Neuromodulation and ablation with focused ultrasound—toward the future of noninvasive brain therapy. Neural Regen Res 14(9):1509–1510
Yoshida M, Kobayashi H, Terasaka S, Endo S, Yamaguchi S, Motegi H, Itay R, Suzuki S, Brokman O, Shapira Y, Moriyama K, Kawase Y, Akahane T, Kato Y, Kamada H, Houkin K (2019) Sonodynamic therapy for malignant glioma using 220-kHz transcranial magnetic resonance imaging-guided focused ultrasound and 5-Aminolevulinic acid. Ultrasound Med Biol 45(2):526–538
Lv Y, Zheng J, Zhou Q, Jia L, Wang C, Liu N, Zhao H, Ji H, Li B, Cao W (2017) Antiproliferative and apoptosis-inducing effect of exo-protoporphyrin IX based sonodynamic therapy on human oral squamous cell carcinoma. Sci Rep 7:40967
Yue W, Chen L, Yu L, Zhou B, Yin H, Ren W, Liu C, Guo L, Zhang Y, Sun L, Zhang K, Xu H, Chen Y (2019) Checkpoint blockade and nanosonosensitizer-augmented noninvasive sonodynamic therapy combination reduces tumour growth and metastases in mice. Nat Commun 10(1):2025
Wan GY, Liu Y, Chen BW, Liu YY, Wang YS, Zhang N (2016) Recent advances of sonodynamic therapy in cancer treatment. Cancer Biol Med 13(3):325–338
Wang X, Jia Y, Wang P, Liu Q, Zheng H (2017) Current status and future perspectives of sonodynamic therapy in glioma treatment. Ultrason Sonochem 37:592–599
Wang X, Jia Y, Su X, Wang X, Zhang K, Feng X et al (2015) Combination of protoporphyrin IX-mediated sonodynamic treatment with doxorubicin synergistically induced apoptotic cell death of a multidrug-resistant leukemia K562/DOX cell line. Ultrasound Med Biol 41:2731–2739
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.
The current research does not involve human participants and/or animals and therefore informed consent was not required.
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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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
- Focused ultrasound
- Sonodynamic therapy