Alternating electric fields transform the intricate network of tumour vasculature into orderly parallel capillaries and enhance the anti-angiogenesis effect of bevacizumab

Abstract

The search for effective strategies to target tumour angiogenesis remains a critical goal of cancer research. We present a pioneering approach using alternating electric fields to inhibit tumour angiogenesis and enhance the therapeutic efficacy of bevacizumab. Chicken chorioallantoic membrane, cell viability and in vitro endothelial tube formation assays revealed that electric fields with a frequency of 1000 kHz and an electric intensity of 0.6 V/cm inhibited the growth of vascular endothelial cells and suppressed tumour-induced angiogenesis. In an animal U87MG glioma model, 1000 kHz electric fields inhibited tumour angiogenesis and suppressed tumour growth. As demonstrated by 3D vessel analysis, tumour vasculature in the control group was a stout, interwoven network. However, electric fields transformed it into slim, parallel capillaries that were strictly perpendicular to the electric field direction. This architectural transformation was accompanied by apoptosis of vascular endothelial cells and a notable reduction in tumour vessel number. Additionally, we found that the anti-angiogenesis and tumour-suppression effects of electric fields synergised with bevacizumab. The anti-angiogenic mechanisms of electric fields include disrupting spindle formation during endothelial cell division and downregulating environmental angiogenesis-related cytokines, such as interleukin-6, CXCL-1, 2, 3, 5 and 8, and matrix metalloproteinases. In summary, our findings demonstrate the potential of alternating electric fields (AEFs) as a therapeutic modality to impede angiogenesis and restrain cancer growth.

References

[1] ShamsS, PatelCB. Anti-cancer mechanisms of action of therapeutic alternating electric fields (tumor treating fields [TTFields]). J Mol Cell Biol. 2022;14(8):mjac047.

[2] MoserJC, Salvador E, DenizK, et al. The mechanisms of action of tumor treating fields. Cancer Res. 2022;82(20):3650-3658.

[3] LeeJ, Grabowski MM, LathiaJD. Tumor treating fields: killing two birds with one stone. J Clin Invest. 2022;132(8):e159073.

[4] NeuhausE, Zirjacks L, GanserK, et al. Alternating electric fields (TTFields) activate Cav1.2 channels in human glioblastoma cells. Cancers (Basel). 2019;11(1):110.

[5] JoY, HanYI, LeeE, et al. The combination of tumor treating fields and hyperthermia has synergistic therapeutic effects in glioblastoma cells by downregulating STAT3. Am J Cancer Res. 2022;12(3):1423-1432.

[6] ChenD, LeSB, HutchinsonTE, et al. Tumor treating fields dually activate STING and AIM2 inflammasomes to induce adjuvant immunity in glioblastoma. J Clin Invest. 2022;132(8):e149258.

[7] KimEH, JoY, SaiS, et al. Tumor-treating fields induce autophagy by blocking the Akt2/miR29b axis in glioblastoma cells. Oncogene. 2019;38(39):6630-6646.

[8] PatelCB, BeinatC, XieY, ChangE, GambhirSS. Tumor treating fields (TTFields) impairs aberrant glycolysis in glioblastoma as evaluated by [18F]DASA-23, a non-invasive probe of pyruvate kinase M2 (PKM2) expression. Neoplasia. 2021;23(1):58-67.

[9] SalvadorE, Köppl T, HörmannJ, et al. Tumor treating fields (TTFields) induce cell junction alterations in a human 3D in vitro model of the blood-brain barrier. Pharmaceutics. 2023;15(1):185.

[10] VoloshinT, KaynanN, DavidiS, et al. Tumor-treating fields (TTFields) induce immunogenic cell death resulting in enhanced antitumor efficacy when combined with anti-PD-1 therapy. Cancer Immunol Immunother. 2020;69(7):1191-1204.

[11] OhJY, LeeYJ, KimEH. Tumor-treating fields inhibit the metastatic potential of osteosarcoma cells. Technol Cancer Res Treat. 2020;19:1533033820947481.

[12] KimEH, SongHS, YooSH, Yoon M. Tumor treating fields inhibit glioblastoma cell migration, invasion and angiogenesis. Oncotarget. 2016;7(40):65125-65136.

[13] GriffioenAW, DudleyAC. The rising impact of angiogenesis research. Angiogenesis. 2022;25(4):435-437.

[14] GarciaJ, Hurwitz HI, SandlerAB, et al. Bevacizumab (Avastin®) in cancer treatment: a review of 15 years of clinical experience and future outlook. Cancer Treat Rev. 2020;86:102017.

[15] LuganoR, Ramachandran M, DimbergA. Tumor angiogenesis: causes, consequences, challenges and opportunities. Cell Mol Life Sci. 2020;77(9):1745-1770.

[16] RoyME, Veilleux C, PaquinA, GagnonA, AnnabiB. Transcriptional regulation of CYR61 and CTGF by LM98: a synthetic YAP-TEAD inhibitor that targets in-vitro vasculogenic mimicry in glioblastoma cells. Anticancer Drugs. 2024;35:709-719.

[17] Laborda-IllanesA, Sánchez-Alcoholado L, Castellano-CastilloD, et al. Development of in vitro and in vivo tools to evaluate the antiangiogenic potential of melatonin to neutralize the angiogenic effects of VEGF and breast cancer cells: CAM assay and 3D endothelial cell spheroids. Biomed Pharmacother. 2023;157:114041.

[18] Al-ZabinA, Abu Thiab T, ZihlifM, et al. Anti-angiogenic and cytotoxic evaluation of green-synthesized Fe2ZnO4 nanoparticles against MCF-7 cell line. Biomed Rep. 2024;20(3):36.

[19] BerkelmannL, BaderA, MeshksarS, et al. Tumour-treating fields (TTFields): investigations on the mechanism of action by electromagnetic exposure of cells in telophase/cytokinesis. Sci Rep. 2019;9(1):7362.

[20] BarnettS, Hellmann F, ParkeE, et al. Vincristine dosing, drug exposure and therapeutic drug monitoring in neonate and infant cancer patients. Eur J Cancer. 2022;164:127-136.

[21] JainRK. Normalization of tumor vasculature: an emerging concept in antiangiogenic therapy. Science. 2005;307(5706):58-62.

[22] NagashimaT, Hadiwidjaja S, OhsumiS, et al. In vitro model of human skeletal muscle tissues with contractility fabricated by immortalized human myogenic cells. Adv Biosyst. 2020;4(11):e2000121.

[23] LeiN, MellemJE, BrockiePJ, Madsen DM, MaricqAV. NRAP-1 is a Presynaptically released NMDA receptor auxiliary protein that modifies synaptic strength. Neuron. 2017;96(6):1303-1316.e6.

[24] BuleP, AguiarSI, Aires-Da-SilvaF, DiasJNR. Chemokine-directed tumor microenvironment modulation in cancer immunotherapy. Int J Mol Sci. 2021;22(18):9804.

[25] BordeleauF, MasonBN, LollisEM, et al. Matrix stiffening promotes a tumor vasculature phenotype. Proc Natl Acad Sci U S A. 2017;114(3):492-497.

[26] JonesSA, Jenkins BJ. Recent insights into targeting the IL-6 cytokine family in inflammatory diseases and cancer. Nat Rev Immunol. 2018;18(12):773-789.

[27] AndersonNM, SimonMC. The tumor microenvironment. Curr Biol. 2020;30(16):R921-R925.

[28] CaoY, LangerR, FerraraN. Targeting angiogenesis in oncology, ophthalmology and beyond. Nat Rev Drug Discov. 2023;22(6):476-495.

[29] OnkenJ, Goerling U, HeinrichM, et al. Patient reported outcome (PRO) among high-grade glioma patients receiving TTFields treatment: a two center observational study. Front Neurol. 2019;10:1026.

[30] ShiW, Blumenthal DT, Oberheim BushNA, et al. Global post-marketing safety surveillance of tumor treating fields (TTFields) in patients with high-grade glioma in clinical practice. J Neurooncol. 2020;148(3):489-500.

[31] NarayananS, Srinivas S. Incorporating VEGF-targeted therapy in advanced urothelial cancer. Ther Adv Med Oncol. 2017;9(1):33-45.

[32] HongP, Kudulaiti N, WuS, NieJ, ZhuangD. Tumor treating fields: a comprehensive overview of the underlying molecular mechanism. Expert Rev Mol Diagn. 2022;22(1):19-28.

[33] CeresoliGL, AertsJG, DziadziuszkoR, et al. Tumour treating fields in combination with pemetrexed and cisplatin or carboplatin as first-line treatment for unresectable malignant pleural mesothelioma (STELLAR): a multicentre, single-arm phase 2 trial. Lancet Oncol. 2019;20(12):1702-1709.

[34] ChangE, Pohling C, BeyguiN, et al. Synergistic inhibition of glioma cell proliferation by Withaferin a and tumor treating fields. J Neurooncol. 2017;134(2):259-268.

[35] JoY, OhG, GiY, et al. Tumor treating fields (TTF) treatment enhances radiation-induced apoptosis in pancreatic cancer cells. Int J Radiat Biol. 2020;96(12):1528-1533.

[36] SalvadorE, Kessler AF, DomröseD, et al. Tumor treating fields (TTFields) reversibly Permeabilize the blood-brain barrier in vitro and in vivo. Biomolecules. 2022;12(10):1348.

[37] ZegeyeMM, Andersson B, SirsjöA, LjungbergLU. IL-6 trans-signaling impairs sprouting angiogenesis by inhibiting migration, proliferation and tube formation of human endothelial cells. Cells. 2020;9(6):1414.

[38] VoloshinT, Schneiderman RS, VolodinA, et al. Tumor treating fields (TTFields) hinder cancer cell motility through regulation of microtubule and acting dynamics. Cancers (Basel). 2020;12(10):3016.

RIGHTS & PERMISSIONS

2024 The Author(s). Cell Proliferation published by Beijing Institute for Stem Cell and Regenerative Medicine and John Wiley & Sons Ltd.