Abstract Background: Amplitude-modulated 27.12 MHz radiofrequency electromagnetic fields (AM RF EMF) delivered via a spoon-shaped antenna placed on the patient’s tongue result in shrinkage of the primary and metastatic tumors in patients with advanced hepatocellular carcinoma (HCC). Treatment is FDA-approved for patients with advanced HCC who fail 1stline and 2ndline therapy (H220001). The mechanism by which AM RF EMF has a direct antiproliferative effect depends on the influx of extracellular Ca2+ into the cancer cell via CaV3.2 (CACNA1H) T-type voltage-gated calcium channel (VGCC). Here we show that HCC-specific AM RF EMF’s (HCCMF) therapeutic action is inhibited by drugs with the same binding profile as ethosuximide. Methods: Computational docking study, using AlphaFold Protein Structure Database (AFDB accession: AF-O95180-F1-v4) showed that ethosuximide binds to an intracellular, hydrophobic pocket formed by amino acids (AAs) Val79 and Phe80 of the free N-terminal segment and Leu160 and Phe161 of S2-S3 linker of CACNA1H Domain 1. To confirm the ethosuximide binding-pocket (EBP) AAs [Val79, Phe80, Leu160, and Phe161] are important for the inhibition of HCCMF, we virtually screened (VS) the FDA library for drugs that would bind similarly to ethosuximide. Drugs predicted / identified via VS where then tested (colony formation, qPCR, and Fluo-4 Ca2+ dye) HCCMF-mediated cell proliferation in HCC cell lines. Results: Tadalafil and abiraterone acetate are the first two drugs found, via VS and tested, to inhibit HCCMF. In the Huh7 cell line, tadalafil blocked HCCMF-mediated cell proliferation with respect to colony formation and Ki-67/Cyclin D1 expression. Tadalafil also blocked Ca2+influx assessed by Fluo-4 in Huh7 and HepG2 cell lines. Interestingly, we found that sildenafil, while belonging to the same class of phosphodiesterase inhibitors as tadalafil, did not block HCCMF-mediated Ca2+influx. This was shown via Fluo-4 Ca2+ staining in both Huh7 and HepG2. Specifically, mean fluorescent intensity (MFI) was higher in the HCCMF treated group even when sildenafil was present. HCCMF-MFI increased by 33.07% (p-value: 0.0162, n=5) in Huh7 and by 31.86% (p-value: 0.0012, n=5) in HepG2 compared to SHAM (no treatment control). Conclusions: The availability of the EBP AAs is necessary for HCCMF-mediated inhibition of HCC cell proliferation. Theoretical modelling confirmed by laboratory experiments demonstrate that binding of drugs to this pocket interferes with Ca2+ influx and block the anti-cancer effects of HCCMF. These results support the use of computational modeling to predict and identify all FDA drugs which could be used during cancer therapy and should thereby be avoided in patients receiving HCCMF treatment. Citation Format: Hugo Jimenez, Ping Dou, Aaban A. Azmi, Elyas Khan, Alhussein Elshebiny, Yusuf Fateh, Nathan Elias, Cameron Osenkowski, Liyue Zhang, Grayson Barker, Allan M. Johansen, Ravi Paluri, Janaka S. Liyanage, Mohammed N. Al Hallak, Anthony F. Shields, Muhammad W. Saif, Husain Y. Khan, Amro Aboukameel, Md Hafiz Uddin, Sahar F. Bannoura, Ramzi M. Mohammad, Alexandre Barbault, Boris C. Pasche. Virtual screening identification of FDA approved drugs blocking the therapeutic action of tumor-specific amplitude-modulated radiofrequency electromagnetic fields [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 5712.

Amplitude-modulated 27.12 MHz radiofrequency electromagnetic fields (AM RF EMF) delivered via a spoon-shaped antenna placed on the patient’s tongue result in shrinkage of the primary and metastatic tumors in patients with advanced hepatocellular carcinoma (HCC). The mechanism by which AM RF EMF has a direct antiproliferative effect has been found to be dependent on the influx of extracellular Ca2+ into the cancer cell via CaV3.2 (gene name CACNA1H) T-type voltage-gated calcium channel. Fibrolamellar Carcinoma (FLC), a rare subtype of HCC, has few options available for treatment. Here we present, for the first time, data showing that HCC-specific AM RF EMF (HCCMF) are effective in FLC. HepG2 is an HCC cell line and H33 (HepG2DNAJB1-PRKACA) was generated by using the HepG2 cell line and generating the DNAJB1-PRKACA gene fusion at the native locus (CRISPR-based 400kbp excision). Cells either received no treatment (SHAM) or were exposed to HCCMF three hours per day for seven days total. HepG2 and H33 cells were exposed using systems replicating human exposure levels and treatment duration. In vitro experiments (colony formation and tumor sphere formation) with HepG2 and H33 cells were performed. Two-tailed Student’s t-test was used for statistical comparison. Proliferation (colony formation assay) of HepG2 and H33 is inhibited by 37.54% (p-value: 0.0023, N=6) and 27.08% (p-value: 0.0006, N=6), respectively. Stemness (tumor sphere formation assay) of HepG2 and H33 is inhibited by 33.17% (p-value < 0.0001, N=8 SHAM and N=7 HCCMF) and 43.95% (p-value< 0.0001, N=7 SHAM and N=8 HCCMF), respectively. Proliferation and stemness of the HepG2 and H33 [HepG2 (DNAJB1-PRKACA)] cell lines were inhibited following HCCMF treatment. Results show the presence of the FLC fusion protein (DNAJB1-PRKACA) does not inhibit the potential of HCCMF as a novel systemic targeted therapy for FLC. This proof-of-principle data shows significant potential for the treatment of FLC. Hugo Jimenez, Liyue Zhang, Grayson Barker, Callum McGrath, Allan M. Johansen, Janaka S. Liyanage, Mohammed N. Al Hallak, Anthony F. Shields, Wasif Saif, Husian Y. Khan, Amro aboukameel, Hafiz Uddin, Sahar F. Bannoura, Ramzi Mohammad, Sean Ronnekleiv-Kelly, Alexandre Barbault, Carl F. Blackman, Asfar Azmi, Boris Pasche. Treatment of fibrolamellar carcinoma with hepatocellular carcinoma -specific amplitude-modulated radiofrequency electromagnetic fields. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 330.

BACKGROUND:Intrabuccal administration of amplitude-modulated 27.12 MHz radiofrequency electromagnetic fields (AM RF EMF) resulting in the systemic delivery of low and safe levels of AM RF EMF has shown activity in several forms of cancer. METHODS:Glioblastoma (GB) cell lines were exposed to GB-specific AM RF EMF (GBMF) three hours per day at a level of exposure identical to patients during treatment. Cellular assays and agnostic genomic approaches were used to characterize the mechanism-of-action. One patient with therapy refractory GB received compassionate use treatment with GBMF as well as a second patient with refractory oligodendroglioma. RESULTS:Treatment with GBMF inhibited the proliferation of several GB cell lines. CACNA1H mediates the effect of GBMF. GBMF modulates the "Mitotic Roles of Polo-Like Kinase" pathway resulting in the disruption of GB mitotic spindle. There was evidence of clinical and radiological benefit in a 38-year-old patient with recurrent GB and evidence of safety and feasibility in a 47-year-old patient with oligodendroglioma. CONCLUSIONS:This is the first report showing in vitro antitumor activity, disruption of the mitotic spindle, activation of the Mitotic Roles of Polo-like kinase pathway in GB. This is also the first report showing feasibility and clinical activity in patients with brain tumor.

613 Background: There are no commercially available devices for the treatment of patients with advanced hepatocellular carcinoma who have failed 1st line and 2nd line therapy. We have identified tumor-specific modulation frequencies in patients with advanced hepatocellular carcinoma. We exposed patients to radiofrequency electromagnetic fields, which are amplitude-modulated from 0.1 Hz to 100 kHz and measured changes in pulse pressure. Frequencies eliciting measurable changes in pulse pressure were selected as tumor-specific frequencies. Methods: Treatment is administered to patients with advanced hepatocellular carcinoma by means of a battery-operated portable device emitting 27 MHz radiofrequency electromagnetic fields, which are amplitude-modulated at hepatocellular carcinoma frequencies. The device is connected to a coaxial cable ending with a spoon-shaped antenna placed on the anterior part of the patient's tongue during treatment. Treatment is administered three times a day for one hour. Results: A total of 69 patients with advanced hepatocellular carcinoma received treatment with the TheraBionic device until progression or death. The median overall survival of patients who received treatment with the TheraBionic device after failing 1st line and 2nd line therapy was 9.15 (95% CI 1.6-34.8) months and the median overall survival of Child-Pugh A patients was 9.5 months, which are comparatively longer than the 7.8 months pooled estimated medians of the placebo arms of 11 randomized 1st line and 2nd line placebo-controlled studies assessing the safety and efficacy of new agents for the treatment of Child-Pugh A advanced HCC (Llovet, Montal et al., J Hepatol 2019). We also analyzed the survival of patients who had received two lines of systemic therapy and did not receive any additional cancer treatment while and after receiving treatment with the TheraBionic device. The median OS of these patients was 9.8 months and the median OS of the Child-Pugh A patients was 17.2 months, which are comparatively longer than the 7.74 months pooled estimated medians of the placebo arms of the above cited 11 randomized studies. To assess unknown device-related adverse events, we assessed patient reported symptoms using the same scale in the SHARP and Asian Pacific Sorafenib studies. The incidence of any grade adverse events among Child-Pugh A who received treatment with the TheraBionic device was not different from the Placebo group of the SHARP and Asian Pacific Sorafenib studies. There were no NCI Grade 2, 3 or 4 toxicities. One patient developed grade 1 mucositis and one patient developed grade 1 fatigue. Conclusions: The data presented here provide reasonable assurance of safety and probable benefit in patients with advanced hepatocellular carcinoma who have failed 1st line and 2nd line therapy: https://www.fda.gov/medical-devices/recently-approved-devices/therabionic-p1-h220001 .

Pancreatic ductal adenocarcinoma (PDAC) is the fourth leading cause of cancer-related mortality, and about 50% of PDC patients present with metastatic disease. Despite advances in systemic therapy, the prognosis for PDC is dismal, with a five-year overall survival (OS) for metastatic PDC of about 5%. The most used regimens for advanced pancreatic cancer include gemcitabine/nab-paclitaxel (Gem/NabP) and FOLFIRINOX, with the recent addition of NALIRIFOX. All these regimens result in median survival rates under one year. Amplitude-modulated 27.12 MHz radiofrequency electromagnetic fields (AM RF EMF) delivered via a spoon-shaped antenna placed on the patient’s tongue results in shrinkage of the primary and metastatic tumors in patients with cancer. The mechanism by which AM RF EMF have direct antiproliferative effect has been found to be dependent on the influx of extracellular Ca2+ into the cancer cell via Cav3.2 (gene name CACNA1H) T-type voltage gated calcium channel. Here we present, for the first time, data showing that Pancreatic cancer-specific AM RF EMF (PancMF) is effective in pancreatic cancer cells. MIAPaCa-2 and PANC-1 cell lines were exposed either to no treatment (SHAM) to PancMF three hours per day for seven days total. Cell lines were exposed using systems replicating human exposure levels and treatment duration. Ethosuximide (Ethos), a pan t-type voltage gated calcium channel (VGCC) blocker was used to inhibit the effects of PancMF. In vitro experiments, (proliferation) Tritiated Thymidine incorporation and (stemness) tumor sphere formation were performed. ANOVA and Two-tailed Student’s t-test were used for comparison. Panc1 cell line: Proliferation was inhibited 38.51% by PancMF treatment (p-value: <0.0001) but in the presence of ethosuximide PancMF proliferation inhibition was blocked (p-value: 0.9897). Stemness was inhibited 41.02% by PancMF treatment (p-value < 0.0001). MiaPaca2 cell line: Proliferation was inhibited 20.67% by PancMF treatment (p-value: 0.0208) but in the presence of ethosuximide PancMF proliferation inhibition was blocked (p-value: 0.8933). Stemness was inhibited 26.54% by PancMF treatment (p-value < 0.0001). Proliferation and Stemness of Panc1 and MiaPaca2 cell were inhibited following PancMF treatment, however, the presence of ethosuximide blocked the anti-proliferative effects of PancMF. Results indicate that the t-type VGCC mediates the anti-proliferative effect. This results trend with previously published data showing that Cav3.2 is the bioantenna and mediator for tumor-specific AM RF EMF treatment. Hugo Jimenez, Callum McGrath, Liyue Zhang, Grayson Barker, Allan M. Johansen, Janaka S. Liyanage, Mohammed N. Al Hallack, Anthony F. Shields, Wasif Saif, Husain Y. Khan, Amro Aboukameel, Hafiz Uddin, Sahar F. Bannoura, Ramzi M. Mohammad, Alexandre Barbault, Carl F. Blackman, Ravi K. Paluri, Asfar Azmi, Boris Pasche. Proliferative inhibition of pancreatic cancer cells treated with pancreatic cancer - specific amplitude-modulated radiofrequency electromagnetic fields. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 329.

Importance : Hepatocellular carcinoma (HCC) is the third leading cause of cancer death worldwide. Despite the recent approval of several new agents, long-term disease control remains elusive for most patients. Administration of 27.12 MHz radiofrequency (RF) electromagnetic fields (EMF) by means of a spoon-shaped antenna (TheraBionic P1 device) placed on the anterior part of the tongue results in systemic delivery of low and safe levels of RF EMF from head to toe. Objective : To report treatment outcomes and adverse events associated with treatment with the TheraBionic P1 device in comparison to suitable historical placebo and actively treated controls. Design : Pooled case series with comparison to historical controls. Participants : Patients with advanced HCC receiving this treatment, 18 real-world patients and 41 patients from a previously reported phase II study. Historical controls from previously conducted clinical trials. Interventions : Three hours daily treatment with the TheraBionic P1 device compared with standard of care as received by historical controls in the previously conducted trials. Main outcomes and measures : Overall survival (OS), time to progression, response rate, and adverse events in the combined pooled patients and in appropriate subgroups comparable to the historical control groups. Results : In the pooled treatment group, median OS of patients with Child-Pugh A disease ( n = 32) was 10.36 (95% CI 5.42–14.07) months, 4.44 (95% CI 1.64–7.13) months for patients with Child-Pugh B disease ( n = 25), and 1.99 (95% CI 0.76–3.22) months for patients with Child-Pugh C disease ( n = 2). Median OS for Child-Pugh A patients was 2.62 (33.9%) months longer than the 7.74 months OS of comparable historical controls ( p = 0.036). The 4.73 (95% CI 1.18–8.28) months median OS for Child-Pugh B patients receiving TheraBionic P1 device as first line therapy is slightly higher than the 4.6 months median OS of historical controls receiving Sorafenib as first line therapy. Only grade 1 mucositis and fatigue were reported by patients using the device, even among Child-Pugh B and C patients. No patients discontinued treatment because of adverse events. Conclusions and Relevance : Treatment of advanced HCC with the TheraBionic P1 device is well tolerated, even in patients with severely impaired liver function, and results in improved overall survival compared to historical controls without any significant adverse events, even after many years of continuous treatment. This treatment modality appears to be well suited for patients who have failed or are intolerant to currently approved therapies.

"Comments on ‘Search for tumor-specific frequencies of amplitude modulated 27 MHz electromagnetic fields in mice with hepatocarcinoma xenografted tumors’." International Journal of Radiation Biology, 96(7), pp. 845–846

BACKGROUND:Administration of amplitude modulated 27·12 MHz radiofrequency electromagnetic fields (AM RF EMF) by means of a spoon-shaped applicator placed on the patient's tongue is a newly approved treatment for advanced hepatocellular carcinoma (HCC). The mechanism of action of tumour-specific AM RF EMF is largely unknown. METHODS:Whole body and organ-specific human dosimetry analyses were performed. Mice carrying human HCC xenografts were exposed to AM RF EMF using a small animal AM RF EMF exposure system replicating human dosimetry and exposure time. We performed histological analysis of tumours following exposure to AM RF EMF. Using an agnostic genomic approach, we characterized the mechanism of action of AM RF EMF. FINDINGS:Intrabuccal administration results in systemic delivery of athermal AM RF EMF from head to toe at levels lower than those generated by cell phones held close to the body. Tumour shrinkage results from differentiation of HCC cells into quiescent cells with spindle morphology. AM RF EMF targeted antiproliferative effects and cancer stem cell inhibiting effects are mediated by Ca2+ influx through Cav3·2 T-type voltage-gated calcium channels (CACNA1H) resulting in increased intracellular calcium concentration within HCC cells only. INTERPRETATION:Intrabuccally-administered AM RF EMF is a systemic therapy that selectively block the growth of HCC cells. AM RF EMF pronounced inhibitory effects on cancer stem cells may explain the exceptionally long responses observed in several patients with advanced HCC. FUND: Research reported in this publication was supported by the National Cancer Institute's Cancer Centre Support Grant award number P30CA012197 issued to the Wake Forest Baptist Comprehensive Cancer Centre (BP) and by funds from the Charles L. Spurr Professorship Fund (BP). DWG is supported by R01 AA016852 and P50 AA026117.

Background: Brain metastases are a major cause of death in patients with metastatic breast cancer. While surgical resection and radiation therapy are effective treatment modalities, the majority of patients will succumb from disease progression. We have developed a novel therapy for brain metastases that delivers athermal radio-frequency electromagnetic fields that are amplitude-modulated at breast cancer specific frequencies (BCF). Methods: 27.12 MHz amplitude-modulated BCF were administered to a patient with a breast cancer brain metastasis by placing a spoon-shaped antenna on the anterior part of the tongue for three one-hour treatments every day. In preclinical models, a BCE dose, equivalent to that delivered to the patient's brain, was administered to animals implanted with either brain metastasis patient derived xenografts (PDXs) or brain-tropic cell lines. We also examined the efficacy of combining radiation therapy with BCF treatment. Additionally, the mechanistic underpinnings associated with cancer inhibition was identified using an agnostic approach. Findings: Animal studies demonstrated a significant decrease in growth and metastases of brain-tropic cell lines. Moreover, BCE treatment of PDXs established from patients with brain metastases showed strong suppression of their growth ability. Importantly, BCF treatment led to significant and durable regression of brain metastasis of a patient with triple negative breast cancer. The tumour inhibitory effect was mediated by Ca2+ influx in cancer cells through CACNA1H T-type voltage-gated calcium channels, which, acting as the cellular antenna for BCF, activated CAMKII/p38 MAPK signalling and inhibited cancer stem cells through suppression of fi-catenin/HMGA2 signalling. Furthermore, BCF treatment downregulated exosomal miR-1246 level, which in turn decreased angiogenesis in brain environment. Therefore, targeted growth inhibition of breast cancer metastases was achieved through CACNA1H. Interpretation: We demonstrate that BCF, as a single agent or in combination with radiation, is a novel treatment approach to the treatment of brain metastases. This paradigm shifting modality warrants further clinical trials for this unmet medical need. (C) 2019 The Authors. Published by Elsevier B.V.

Cancer treatment and treatment options are quite limited in circumstances such as when the tumor is inoperable, in brain cancers when the drugs cannot penetrate the blood-brain-barrier, or when there is no tumor-specific target for generation of effective therapeutic antibodies. Despite the fact that electromagnetic fields (EMF) in medicine have been used for therapeutic or diagnostic purposes, the use of non-ionizing EMF for cancer treatment is a new emerging concept. Here we summarize the history of EMF from the 1890's to the novel and new innovative methods that target and treat cancer by non-ionizing radiation.

Whole body administration of low-level radiofrequency electromagnetic fields, amplitude-modulated (AM RF EMF) at specific frequencies ranging from 400 Hz to 21 kHz showed efficacy in advanced breast cancer and hepatocellular carcinoma (HCC) (Barbault, Costa et al., 2009, Costa de Oliveira et al., 2011). In vitro studies demonstrated that AM RF EMF treatment inhibits cancer cell growth in a tumor-specific manner (Zimmerman, Pennison et al., 2012). Indeed, the growth of both HCC (HepG2 and Huh7) and breast cancer (MCF-7) cells was inhibited when exposed to corresponding tumor-specific frequencies for 3 hours per day for 7 days. The GBM cell line U-251 as well as two glioblastoma (GBM) low passage explant cells (BTCOE-4536 and BTCOE-4795) were exposed to 27.12 MHz RF EMF modulated at GBM-specific frequencies using exposure systems designed to replicate human exposure levels. Control cells were not exposed to RF EMF. GBM cells were cultured for seven days receiving GBM-specific treatment for three hours daily. The specific absorption rate (SAR) of AM RF EMF was 0.4 W/kg, which is identical to the highest SAR measured in patients receiving treatment with this therapy. Proliferation was quantified via the tritiated thymidine incorporation assay. Neurosphere-formation was counted at 7 days post treatment. Statistics were performed by student’s T-test. The growth of U-251, BTCOE 4536 and BTCOE 4795 cell lines was decreased by 19% (N=10; p=0.0018), 23% (N=4; p=0.0348) and 14.5% (N=10; p=0.0431) following exposure to tumor-specific AM RF EMF. Neurosphere-forming ability of the U-251 and BTCOE-4795 cell lines was inhibited by 48.0% (N=6; p=0.0040) and 31.7% (N=6; p=0.0002) following exposure to tumor-specific AM RF EMF. GBM-specific AM RF EMF inhibits proliferation of three different GBM cell lines and neurosphere-forming ability which suggests that AM RF EMF should be further studied as a potentially new treatment modality of GBM.

11079 Background: Experimental and clinical evidence shows that whole body administration of low-level radiofrequency electromagnetic fields, amplitude-modulated (AM RF EMF) at specific frequencies ranging from 400 Hz to 21 kHz has efficacy in advanced hepatocellular carcinoma (HCC) (Costa, de Oliveira et al, 2011). In vitro studies demonstrate that AM RF EMF treatment inhibits cancer cell growth in a tumor-specific manner (Zimmerman, Pennison et al, 2012). RNA-seq analysis of HepG2 cells treated with AM RF EMF revealed altered expression of several mRNAs belonging to the IP3/DAG signaling pathway. Given the central modulatory effect of calcium in this pathway, we hypothesized that calcium modulates the growth inhibitory effect of RF EMF. Methods: HCC and ovarian cancer cell lines as well as NOD SCID mice, carrying subcutaneous HCC xenografts, were exposed to 27.12 MHz RF EMF modulated at cancer-specific frequencies or random chose frequencies using exposure systems designed to replicate human exposure levels. Huh-7 cells were cultured in the presence or absence of BAPTA, a Ca2+ chelator, for three hours daily, seven days in a row. Mice carrying Huh-7 xenografts received treatment three hours daily for six weeks. The specific absorption rate (SAR) of AM RF EMF was 0.4 W/kg, which is identical to the highest SAR measured in patients receiving treatment with this therapy. mRNA expression of key mediators of the IP3/DAG signaling pathway was analyzed. Results: Ca2+ chelation fully abrogated AM RF EMF inhibition of HCC cell growth. ARHGDIB and S100B mRNA was down regulated in Huh-7 cells treated with cancer-specific AM RF EMF. In vivo Huh-7 was significantly inhibited in mice exposed to AM RF EMF. Conclusions: Ca2+ is a necessary mediator of AM RF EMF anti-proliferative effects in HCC, which modulate the IP3/DAG signaling pathway (Costa, F. P., et al, 2011.

e15155 Background: Werecently described the antiproliferative effects of low levels of electromagnetic fields, amplitude-modulated at disease-specific frequencies (AM EMF) in patients with HCC (Br J Cancer, 2011,105:640) and on HCC cells (Br J Cancer, 2012,106:307). We identified HCC-specific modulation frequencies using a patient based biofeedback approach (J Exp Clin Cancer Res, 2009,28:51). Specific frequencies cause changes in pulse pressure in patients with HCC but not in healthy individuals. The effects of AM EMF in HBsAg+ patients, a pre-malignant condition, are unknown. Methods: 17 healthy individuals and 17 patients (5 HCC and 12 HBsAg+) were exposed to 206 HCC-specific modulation frequencies ranging from 100 Hz to 21 kHz (HCC-frequencies) and 237 random chosen frequencies (non-specific frequencies) within the same frequency range. All participants were exposed to both AM EMF and each frequency was tested separately. Changes in pulse pressure were recorded as previously described (J Exp Clin Cancer Res, 2009, 28:51). Results: HCC-frequencies caused changes in pulse pressure in all HBsAg+ and patients with HCC but no changes were observed in healthy individuals. The median number of frequencies causing changes in pulse pressure in patients with HCC was 175 (range: 97-206) and 7 (range: 3-16) in HBsAg+ patients. The non-specific frequencies did not cause changes in pulse pressure in patients with HCC but it caused changes in pulse pressure in 9 HBsAg+ (75%) with a median number of responding frequencies of 2 (range: 1-6). Non-specific frequencies also caused changes in pulse pressure in 11 healthy individuals (64%) with a median number of responding frequencies of 11 (range: 1-15). The pattern of changes in pulse pressure were significant between the 3 groups (p=0.0001). Conclusions: EMF modulated at specific frequencies cause a significantly different pattern of changes in pulse pressure in the three groups. These findings provide support for the development of EMF as a new diagnostic procedure. Clinical trial information: NCT01641276.

Abstract Purpose: Hepatocellular carcinoma (HCC) incidence in the US is dramatically increasing. Five-year survival remains 3-5%, demonstrating urgent need for additional therapies. Intrabuccal administration of amplitude modulated electromagnetic fields (RF EMF) is a novel, minimally invasive treatment modality which results in whole body absorption of very low levels of RF EMF. Clinical studies show that this treatment approach elicits therapeutic responses in patients with hepatocellular carcinoma and breast cancer. Using an in vitro exposure system replicating the levels of exposure achieved in humans, we have described a phenotype in HCC cells following RF EMF exposure that included proliferative inhibition, modulation of gene expression, and disruption of the mitotic spindle. This phenotype was specific for HCC cells exposed to HCC-specific RF EMF at exposure levels ranging from 0.03 to 0.4 W/kg. Methods: HCC cells were exposed to 27.12 MHz electromagnetic fields modulated at specific frequencies from 400 Hz to 21 kHz, previously identified in HCC patients. MicroRNA arrays compared exposed and control groups of HCC cells, with microRNA validation followed by Western blot of target genes and proteins. HCC xenografts were injected subcutaneously in NOD SCID mice. Following palpable tumor establishment, mice were exposed to HCC-specific RF EMF at a specific absorption rate of 0.4 W/kg, euthanized following excessive tumor burden, and evaluated by immunohistochemistry. Results: We identified increased levels of miRNAs targeting proteins belonging to the PI3K pathway, specifically IP3/DAG signaling and intracellular calcium release. This pathway is frequently disrupted in HCC and breast cancer, making it an excellent candidate for modulation by RF EMF; furthermore, downstream effects include: cell cycle progression, proliferation, inhibition of apoptosis, and cell migration. We observed tumor shrinkage in mice exposed to HCC-specific modulation frequencies and residual xenograft tumor cells were infiltrated with fibrous tissue and showed significantly decreased proliferation and increased apoptosis. There was no evidence of altered cell proliferation or fibrosis in other organs. Conclusion: These findings are the first evidence of the efficacy and safety of RF EMF in HCC using a subcutaneous xenograft model and uncover a novel mechanism that controls cancer cell growth in vivo at specific modulation frequencies, possibly through modulation of PI3K signaling and downstream release of intracellular calcium. Citation Format: Hugo Jimenez, Jacquelyn W. Zimmerman, Michael J. Pennison, Ivan Brezovich, Nengjun Yi, Celeste T. Yang, Ryne Ramaker, Devin Absher, Richard M. Myers, Niels Kuster, Frederico P. Costa, Alexandre Barbault, Boris Pasche. Cancer cell proliferation is inhibited by specific modulation frequencies. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5612. doi:10.1158/1538-7445.AM2013-5612 Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.

In the past century, there have been many attempts to treat cancer with low levels of electric and magnetic fields. We have developed noninvasive biofeedback examination devices and techniques and discovered that patients with the same tumor type exhibit biofeedback responses to the same, precise frequencies. Intrabuccal administration of 27.12 MHz radiofrequency (RF) electromagnetic fields (EMF), which are amplitude-modulated at tumor-specific frequencies, results in long-term objective responses in patients with cancer and is not associated with any significant adverse effects. Intrabuccal administration allows for therapeutic delivery of very low and safe levels of EMF throughout the body as exemplified by responses observed in the femur, liver, adrenal glands, and lungs. In vitro studies have demonstrated that tumor-specific frequencies identified in patients with various forms of cancer are capable of blocking the growth of tumor cells in a tissue- and tumor-specific fashion. Current experimental evidence suggests that tumor-specific modulation frequencies regulate the expression of genes involved in migration and invasion and disrupt the mitotic spindle. This novel targeted treatment approach is emerging as an appealing therapeutic option for patients with advanced cancer given its excellent tolerability. Dissection of the molecular mechanisms accounting for the anti-cancer effects of tumor-specific modulation frequencies is likely to lead to the discovery of novel pathways in cancer.

e15049 Background: The incidence of hepatocellular carcinoma (HCC) is increasing but five-year survival remains dismal. Intrabuccal administration of amplitude-modulated radiofrequency electromagnetic fields (RF EMF) is a novel, non-invasive treatment modality resulting in whole body absorption of very low levels of RF EMF. Recent studies show that this approach elicits radiological responses in patients with HCC and breast cancer (J Exp Clin Cancer Res, 2009, 28:51; Br J Cancer, 2011, 105:640). Using an in vitro exposure system replicating the levels of exposure achieved in humans, we have shown that HCC cells and breast cancer cells are growth inhibited by HCC-specific and breast cancer-specific frequencies, respectively. Additionally, RF EMF exposure causes modulation of gene expression and disruption of the mitotic spindle (Br J Cancer 2012, 106:307). Methods: HCC cells were exposed to radiofrequency electromagnetic fields modulated at specific frequencies previously identified in HCC patients. MicroRNA arrays compared exposed and control groups of HCC cells. HCC cells were injected subcutaneously in NOD SCID mice. Following palpable tumor establishment, mice were exposed to HCC-specific RF EMF at a specific absorption rate of 0.4 W/kg and euthanized following excessive tumor burden. Results: We identified increased levels of miRNAs targeting proteins belonging to the PI3K pathway, specifically IP3/DAG signaling and intracellular calcium release, a pathway frequently disrupted in HCC and breast cancer. While HCC xenografts grew in control mice, we observed significant tumor shrinkage in mice exposed to HCC-specific modulation frequencies and residual xenograft tumor cells were infiltrated with fibrous tissue and showed significantly decreased proliferation and increased apoptosis. There was no evidence of altered cell proliferation or fibrosis in other organs. Conclusions: These findings are the first in vivo evidence of the efficacy of RF EMF in HCC and uncover a novel mechanism that targets cancer cell growth at specific modulation frequencies, by means of changes in PI3K signaling and release of intracellular calcium.

Abstract Background: Hepatocellular carcinoma (HCC), a leading cause of cancer death worldwide, now has the fastest growing rate in the United States. The intrabuccal administration of amplitude modulated electromagnetic fields (AM-EMF) is a novel, minimally invasive treatment modality. There is clinical evidence that this novel treatment approach elicits therapeutic responses in patients with cancer (1, 2). However, there is no known mechanism explaining the anti-proliferative effect of very low intensity electromagnetic fields. Methods: Hepatocellular carcinoma cells were exposed to 27.12 MHz radiofrequency electromagnetic fields using in vitro exposure systems designed to replicate in vivo conditions. Cancer cells were exposed to tumor-specific modulation frequencies, previously identified by biofeedback methods in patients with a diagnosis of cancer. Control modulation frequencies consisted of randomly-chosen modulation frequencies within the same 100 Hz to 21 kHz range as cancer-specific frequencies. Growth inhibition was evaluated by measuring tritiated thymidine incorporation. RNA-Seq was used to identify genes with differential expression in cells exposed to HCC-specific AM-EMF. Confocal laser scanning microscopy allowed the visualization of the mitotic spindle. Karyotype assessment compared HepG2 receiving HCC-specific AM-EMF to those not receiving exposure. Results: The growth of hepatocellular carcinoma and breast cancer cells was significantly decreased by hepatocellular carcinoma-specific and breast cancer-specific modulation frequencies, respectively. However, the same frequencies did not affect proliferation of nonmalignant hepatocytes or breast epithelial cells. Inhibition of hepatocellular carcinoma cell proliferation was associated with downregulation of XCL2 and PLP2. Furthermore, hepatocellular carcinoma-specific modulation frequencies disrupted the mitotic spindle. Karyotype analysis was unrevealing. Conclusion: These findings uncover a novel mechanism controlling the growth of cancer cells at specific modulation frequencies without affecting normal tissues, which may have broad implications in oncology. Reference List (1) Barbault A, Costa F, Bottger B, Munden R, Bomholt F, Kuster N, et al. Amplitude-modulated electromagnetic fields for the treatment of cancer: Discovery of tumor-specific frequencies and assessment of a novel therapeutic approach. Journal of Experimental & Clinical Cancer Research 2009;28:51. (2) Costa FP, de Oliveira AC, Meirelles R, Machado MCC, Zanesco T, Surjan R, et al. Treatment of advanced hepatocellular carcinoma with very low levels of amplitude-modulated electromagnetic fields. Br J Cancer 2011;105:640-8. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 916A. doi:1538-7445.AM2012-916A

e15573 Background: Over the past few years we have identified tumor-specific frequencies for several common forms of cancer. The goal of this study was to assess the tolerability and effectiveness of electromagnetic fields amplitude-modulated at tumor-specific frequencies and administered by means of an intrabuccal spoon-shaped probe in patients with advanced hepatocellular carcinoma (HCC). Methods: From October 2005 to July 2007, patients with advanced HCC and Child-Pugh A or B were recruited in a phase II study. Three daily 60 min outpatient treatments were administered until disease progression or death. Imaging studies were performed every eight weeks. The primary efficacy end point was progression-free survival ≥ 6 months. Secondary efficacy end points were progression-free survival and overall survival. Results: A total of 41 patients were enrolled, 17 had Child-Pugh A, 20 Child-Pugh B disease. The median age was 64.0 years. Seventeen patients (34.1%) were progression-free for more than 6 months. Median progression-free and overall survivals were 4.8 months (95% CI 2.3–6.0) and 6.9 months (95 CI 4.8–11.1). As of December 2008, four patients are alive and two patients, who are still undergoing therapy, remain progression-free for 30.4 and 30.7 months, respectively. Four patients had partial response (9.8%) and sixteen had stable disease for at least 12 weeks (39.0%) according to the RECIST criteria resulting in 48.8% disease control. All responses were confirmed by independent review. There were no NCI grade 2, 3 or 4 toxicities. One patient developed grade 1 mucositis and one patient grade 1 fatigue. Conclusions: In patients with advanced HCC and impaired hepatic function, treatment with amplitude-modulated electromagnetic fields is safe, well tolerated, and shows evidence of anti-tumor effects, which are long-lasting in some patients. No significant financial relationships to disclose.