Absracts of the 39th Conference of the International Clinical Hyperthermia Society
It’s time for prime time for oncologic hyperthermia/thermotherapy ?
Stephan Bodis 1-2-3, Oliver Riesterer 1, Emsad Puric 1, Niloy Datta 4
1 Kantonsspital Aarau
2 University Hospital Zurich
3 ITIS Foundation Zurich
4 Mahatma Gandhi Institute, Sevagram, Wardha, Maharashtra, India, Pin – 442102
Scientific evidence and the road towards public acceptance of regional, low temperature, non-ablative oncologic hyperthermia/thermotherapy combined with fractionated state of the art radiotherapy is bumpy and reflects to some extent the history of radiation oncology in the 70s. (e.g. Discrepancy in reported results from often small single center phase II trials; lack of phase III multicentric, international trials with long term f/u; establishment and acceptance of a common interest platform for all stakeholders from industry, medicine, patient advocacy groups and science; a common language/dictionary understood and accepted by all stakeholders).
To keep things simple, we postulate 7 bullet points likely to be most relevant to make hyperthermia/thermotherapy combined with RT prime time within the next 5 years:
1) We need more evidence based medecine
2) We need multicentric, prospective, randomised, international clinical trials
3) We need adherence to international, published guidelines (e.g. ESHO guidelines for superficial und deep hyperthermia) and an ISO-certification for clinical hyperthermia units to guarantee a minimal QA standard
4) We need (to strenghthen existing) clinical international research networks in biology, clinics, physics and technology
5) We need national hyperthermia re-imbursement for defined oncology indications in the curative and palliative setting
6) We need a better standardisation of existing and new hard-and software
7) We need better standardisation of patient workflow and QA for both hyperthermia and the combination of HT and RT
Hyperthermia: A potential game changer in the management of cancers in low-middle-income group countries
Niloy Ranjan Datta1, Bharati Mahindrakar Jain1, Zatin Mathi1, Sneha Datta2, Satyendra
Johari3, Ashok Ramji Singh1, Pallavi Kalbande1, Pournima Kale1, Vitaladevuni Shivkumar4,
1 Department of Radiotherapy, Mahatma Gandhi Institute of Medical Sciences, Sevagram, Wardha,
2 Animal Production and Health Laboratory, Joint FAO/IAEA Division of Nuclear
Techniques in Food and Agriculture, Department of Nuclear Sciences and Applications, International
Atomic Energy Agency (IAEA), Vienna, Austria,
3 Johari Digital Healthcare Limited, Jodhpur, India,
4Department of Pathology, Mahatma Gandhi Institute of Medical Sciences, Sevagram, Wardha,
5Foundation for Research on Information Technologies in Society (IT’IS), Zurich,
Loco-regional hyperthermia at 40°C – 44°C is a multifaceted therapeutic modality with a triple advantage of being a potent radiosensitizer, a chemosensitizer and an immunomodulator. Risk difference estimates from pairwise meta-analysis have shown that the local tumour control could be improved by 22.3% (p<0.001), 22.1% (p<0.001) and 25.5% (p<0.001) in recurrent breast cancers, locally advanced cancer cervix (LACC) and locally advanced head neck cancers respectively by adding hyperthermia to radiotherapy over radiotherapy alone. Furthermore, thermochemoradiotherapy in LACC has shown to reduce the local failure rates by 10.1% (p=0.03) and decrease deaths by 5.6% (95% CI: 0.6% – 11.8%) over chemoradiotherapy alone. As around one-third of the cancer cases in lowmiddle-income group countries belong to breast, cervix and head neck regions; hyperthermia could be a potential game changer and expected to augment the clinical outcomes of these patients in conjunction with radiotherapy and/or chemotherapy. Further, hyperthermia could also be a costeffective therapeutic modality as the capital costs for setting up a hyperthermia facility is relatively low. The positive outcomes evident from various phase III randomized trials and meta-analysis with thermoradiotherapy or thermochemoradiotherapy justifies the integration of hyperthermia in the therapeutic armamentarium of clinical management of cancer, especially in low-middle-income group countries.
Use of the preclinical system LabEHY-200 to demonstrate the enhancing anticancer effects of amplitude modulation
Peter Wust1, Danai P. Veltsista1, Stefan Meinke2, Eva Oberacker1, Dennis Kobelt3, 4, 5, Marcus Beck1, Wolfgang Walther3, Jacek Nadobny1, Anja Sterner-Kock3, Florian Heyd2, Ulrike Stein3, 4, 5, Pirus Ghadjar1*
1Department of Radiation Oncology, Charité – Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Augustenburger Platz 1, 13353 Berlin, Germany
2Institute of Chemistry and Biochemistry, Freie Universität Berlin, Laboratory of RNA Biochemistry, Takustrasse 6, 14195 Berlin, Germany
3Experimental and Clinical Research Center, Charité – Universitätsmedizin Berlin and Max Delbrück Center for Molecular Medicine in the Helmholtz Association, Robert-Rössle-Str. 10, 13092 Berlin, Germany
4Max Delbrück Center for Molecular Medicine in the Helmholtz Association, AG Translational Oncology of Solid Tumors, Berlin, Germany
5German Cancer Consortium (DKTK), Im Neuenheimer Feld 280, 69120 Heidelberg, Germany
*Corresponding author: Prof. Dr. Pirus Ghadjar, Charité Universitätsmedizin Berlin – Department of Radiation Oncology,Augustenburger Platz 1, 13353 Berlin, Germany, Phone: +49-30-450657055,
The existence of non-thermal effects of radiofrequency electromagnetic fields (RF-EMF) remains largely elusive. We applied the preclinical system LabEHY-200 at 13.56 MHz plus/minus amplitude modulation at ≥ 100 Hz (1/f spectrum) with dedicated applicators for in vitro and in vivo studies. Our experiments demonstrated that additional amplitude modulation increases the non-thermal effects of RF-EMF in a colorectal cancer (CRC) model using HT-29 cells. Modulated fields promote cell death in CRC cells in vitro as compared to temperature increase alone. This was confirmed in a xenograft mouse model in vivo where tumor growth was inhibited most strongly by modulated fields and less by RF-EMF or temperature alone. Physical parameters and power density are also different and might time-efficiently predict biological endpoints. Our findings suggest that amplitude modulation enhance non-thermal anticancer effects of unmodulated RF-EMF at 13.56 MHz.
Update on mEHTGLIO Phase III trial. First results and comments
Elisabeth Arrojo1, Paola Navarrete1, Uriel Corro1
1University Hospital Marques de Valdecilla, Santander, Spain
Introduction: High grade glioma is a very serious cancer disease with a extremely short life expectancy. Modulated electro-hyperthermia (mEHT) may improve tumor control and survival without significant toxicities in patients with high grade glioma. No randomized trials combining mEHT + standard treatment (ST) with radiotherapy (RT) + chemotherapy (CT) vs ST alone in this type of patients have been published.
Material and methods: Patients diagnosed with WHO grade III/IV glioma at University Hospital Marques of Valdecilla were recruited in phase III mEHTGlio trial between August 2019 and March 2021. The trial has two arms:
- Arm 1: Patients at first diagnosis randomized to control group (C) with ST vs experimental group (E) with ST+mEHT.
- Arm 2: Patients with progression after ST, treated with mEHT in monotherapy or concomitant with CT.
Due to COVID19’s pandemic and technical reasons recruitment was stopped for 18 months during this period.
Results: Twenty-six patients were recruited (14 Arm 1, 12 Arm 2). There were no significant toxicities from mEHT in any of the groups. Average overall survival in Arm 2 was 16,7 months with an average follow-up of 23 months (7-43). Regarding Arm 1, 8 patients were randomized to E group and 6 to C group. 4 patients were excluded for this report: 3 did not begin/complete treatment (2E and 1C group), 1 was a grade III glioma (E group). All the other patients (C y E) were Grade IV (GBM). Due to the low number of patients recruited and in order to make more comparable groups the grade III patient was excluded.
N (Patients evaluated)
17% Positive (n=1)
17% Positive (n=1)
22,09 months (8,8-28,4)
27,99 months (22,10-30,4)
CONCLUSIONS: These are the first preliminary results from a phase III trial comparing ST vs ST+mEHT in patients diagnosed with high grade glioma. COVID19 pandemic has delayed research and we don’t have enough number of patients to rise conclusions but both groups seem to be comparable and the E group seems to have better results regarding overall survival. Adding mEHT to RT/CT did not increase toxicities. More patients and follow-up is needed to rise conclusions.
Enhancing surface electromagnetic intensity by covering a silver fabric for superficial located tumor treatment during oncothermia
Kwan-Hwa Chi, M.D.
The water bolus electrode benefits surface cooling, the body surface close-contacting, and avoiding the edge overheating. However, it might decrease the intensity of electromagnetic (EM) current from metal electrodes. We have previously reported that large and superficially located tumors had better treatment results than deeply seated tumors by oncothermia. We hypothesized that covering a thin silver fabric over the tumor site between the water bolus and body surface might gain more EM current. Textile lined with silver fiber has been developed for protection from EM waves. The interference of EM current may be absorbed and then re-irradiated from silver fabrics as a repeater. (Textile-based generator) The idea was tested in muscle tissue-equivalent agar phantom and three breast cancer patients with Oncotherm EHT-2000 system. The temperature measured by thermocouples inserted in a 15cm catheter and the temperature profiles ±6cm from center, 3cm below the surface of the phantom, was plotted under a 20cm diameter electrode with or without 8cm diameter silver fabric with the power of 60W.
One primary breast cancer and another two locally recurrent breast cancer patients were enrolled and treated with weekly oncothermia combined with radiotherapy and oral chemotherapy with low dose Xelode and Navelbin. All three patents used 20cm diameter electrodes with silver fabrics. Two patients had complete responses, and one had a good partial response, including the distant abscopal effect. She had combined treatment with low-dose immune checkpoint inhibitors.
The power absorption at 3cm below the surface of the phantom shall be regarded as the temperature measured. The temperature was higher for 0.3℃ to 0.6℃ after the application of silver fabric. Interestingly, the temperature profiles seemed to be more homogeneous by adding silver fabric. The skin surface under silver material has a similar temperature as measured by direct temperature measurement, but it was higher by infrared thermometry. Patients felt hotter over the skin but expressed a little discomfort during the highest power of treatment.
The reflected power read from the standing wave ratio (SWR) was always less the 1.06 during treatment. The results of this study may promote the optimal use of Oncomtermia for superficially located tumors. The silver textile may reduce the hot/cold spot for a more realistic inhomogeneous situation. The silver fabric can quickly follow the complexed skin contours in a patient with tumor eruption out of the skin.
An overview of Oncothermia as a treatment modality for cervical cancer
Carrie Anne Minnaar1,2, Jeffrey Kotzen1,2
1 Wits Donald Gordon Medical Centre; Radiation Oncology
2 University of the Witwatersrand; Radiation Sciences
Objective: Modulated electro-hyperthermia (mEHT) is a mild heating technique, using a capacitive-coupled set-up to transmit amplitude modulated 13.56MHz radiofrequency waves between two electrodes. We summarise the literature on mEHT for the management of cervical cancer, describe a cost effectiveness analysis (CEA) on mEHT plus radiotherapy (RT) for the management of locally advanced cervical cancer (LACC), and report on the new three year survival data from the ongoing randomised controlled Phase III trial on mEHT plus chemoradiotherapy (CRT) in South Africa (SA).
Materials and methods: A literature search for “hyperthermia”, “modulated electro-hyperthermia”, and “Oncothermia” in “oncology”, and “cervical cancer” was conducted. Studies that did not utilise mEHT were excluded. All papers on mEHT used for the management of cervical cancer were included. For the CEA, an economic analysis was done with a time horizon of three years, and from the perspective of a third party payer, using three year data from the Dutch Deep HT trial , extrapolated into the SA setting. The costs are reported in SA Rands (ZAR; 2012 value). This model did not assign costs to dying which gives the least effective treatment a cost benefit. The cost effectiveness Markov model, with six months cycle length, considered direct medical costs only, with the primary outcome being Cost per Quality Adjusted Life Year (QALY). Data from the ongoing LACC SA trial were used to evaluated three year survival for patients treated with mEHT plus CRT.
Results: In 2017 Lee et al. reported on a randomised trial evaluating the effect of mEHT (three times/week for a total of 36 treatments), combined with platinum based chemotherapy (ChT; n=18) compared with ChT alone (n=20) for recurrent/residual cervical cancer that had been previously irradiated. Patients with loco-regional metastases were included. Power was set at 80W for the first 10 minutes, 120 W for the next 10 minutes and 150W for the remaining mEHT treatment time. The overall response (complete remission + partial remission + stable disease/progressive disease) was significantly higher in the ChT+mEHT group than in the ChT group (p=0.0461), and this difference remained significant at the last follow-up visit (p=0.0218), although the difference in survival between the two groups was not significant (p=0.235). mEHT did not result in any differences in treatment toxicity . Lee et al. also reported on the improved blood perfusion of cervical cancer tumours after the administration of mEHT, confirming the basis for prescribing mEHT as a radiosensitiser for these tumours . Minnaar et al. have published three papers on the ongoing mEHT trial in which they reported improved local disease control (LDC) with the addition of mEHT to CRT protocols (45.5% versus 24.1%; p=0.003) for the management of LACC in a resource-constrained setting in South Africa , without any significant effect on early toxicity . In the study, mEHT was administered twice per week, immediately before external beam radiation (EBRT) for 55 minutes, starting at 60W and increasing to 130W over five minutes. An abscopal effect was reported in participants in whom extra-pelvic nodal disease was visualized on the pre-treatment 18F-FDG PET/CT studies , confirming the immune-modulating effects of mEHT described in pre-clinical studies. For the CEA, a base case analysis showed that the addition of mEHT to RT dominated treatment by RT alone, and that the addition of mEHT was less costly and more effective. This result was driven by the difference in progression free survival, due to the high costs of progressive disease, and a probabilistic sensitivity analysis confirmed the results of the base case analysis. In the ongoing trial in SA, 98 out of 106[93%] and 99 out of 104[95%] participants in the mEHT and control groups respectively, were available for evaluation at three years post-treatment. The mEHT group was significantly more likely to achieve three year all mortality survival and disease free survival (DFS) at three years than the control group (mEHT: n=48[49%] vs Control: n=38[38%]; HR: 1.45; 95%CI: 1.0-2.1; p=0.044 and mEHT: n=33[34%] vs Control: n=14[14%]; p=0.001; OR:2.4; 95%CI: 1.3-4.4; p=0.003 respectively).
Conclusion: Local recurrences and local disease progression from LACC are associated with morbidity and poor quality of life and the use of mEHT combined with ChT for the management of residual or recurrent disease significantly improves local disease response in these patients. mEHT also improves LDC, three year survival, and three year DFS without increasing the toxicity profile of CRT for LACC patients. Following the review, we recommend mEHT be included in the guidelines for the management of LACC and recurrent/residual cervical cancer. A CEA analysis of mEHT plus CRT using the new three year survival data is underway.
- Van Der Zee J, González González D. The Dutch Deep Hyperthermia trial: Results in cervical cancer. International Journal of Hyperthermia. 2002;18(1):1-12.
- Lee S, Lee N, Cho D, Kim J. Treatment outcome analysis of chemotherapy combined with modulated electro-hyperthermia compared with chemotherapy alone for recurrent cervical cancer, following irradiation. Oncology Letters. 2017;14(1):73-78.
- Lee S-Y, Kim J-H, Han Y-H, Cho D-H. The effect of modulated electro-hyperthermia on temperature and blood flow in human cervical carcinoma. International Journal of Hyperthermia. 2018;34(7):953-960.
- Minnaar CA, Kotzen JA, Ayeni OA, et al. The effect of modulated electro-hyperthermia on local disease control in HIV-positive and -negative cervical cancer women in South Africa: Early results from a phase III randomised controlled trial. PLoS ONE. 2019;14(6):1-23.
- Minnaar CA, Kotzen JA, Naidoo T, et al. Analysis of the effects of mEHT on the treatment- related toxicity and quality of life of HIV-positive cervical cancer patients. International Journal of Hyperthermia. 2020;37(1):263-272.
- Minnaar CA, Kotzen JA, Ayeni OA, Vangu M, Baeyens A. Potentiation of the Abscopal Effect by Modulated Electro-Hyperthermia in Locally Advanced Cervical Cancer Patients. Frontiers in Oncolgy. 2020;10:Article 376.
Advanced Pancreatic cancer treated with combo therapy (modulated electro hyperthermia and chemotherapy): a new hope?
Giammaria Fiorentini1, Donatella Sarti2, Girolamo Ranieri3, Cosmo Damiano Gadaleta3, Caterina Fiorentini4, Carlo Milandri5, Andrea Mambrini6, Stefano Guadagni7
1 Oncology, Poliambulatorio Esculapio, via Vincenzo Casciarolo 9 / A, 40127-Bologna.
2 Oncology Department, Ospedale S. Maria Della Misericordia, ASUR1, Urbino 61029, Italy
3 Interventional and Integrated Medical Oncology, National Cancer Research Centre, IRCCS Istituto Tumori “Giovanni Paolo II”, Bari 70124, Italy
4 Department of Medical Biothecnologies, Division of Cardiology, University Hospital of Siena, Siena 53100, Italy
5 Medical Oncology Unit, San Donato Hospital, Arezzo 52100, Italy
6 Oncology Department, Apuane Hospital, Massa-Carrara 54100, Italy
7 Applied Clinical Sciences and Biotechnology, Section of General Surgery, University of L’Aquila, L’Aquila 67100, Italy
Background: An increasing number of studies report the beneficial effects of regional hyperthermia in association to chemotherapy and radiotherapy for the treatment of pancreatic cancer, in particular, the use of modulated electro-hyperthermia (mEHT) results in increased survival and tumor response.
Aim: To compare outcomes of chemotherapy (CHT) alone or in association with mEHT for the treatment of stage III and IV pancreatic cancer.
Methods: This was an observational retrospective study, data were collected for patients with stage III-IV pancreatic cancer that were treated with CHT alone or in combination with mEHT from 2003 to 2019. A total of 158 patients were included in the study out 270 patients screened in 4 Italian hospitals, 58 (37%) of these received CHT+ mEHT and 100 (63%) CHT. CHT was mainly gemcitabine-based regimens in both groups.
Results: Overall (19.5 mo vs 11.02 mo, P < 0.001) and progression free (12 mo vs 3 mo, P < 0.001) survival were better for the CHT+mEHT group. The association of mEHT resulted also in an improvement of tumor response with disease control rate 95% vs 58 % (P < 0.001) at three months. Toxicity was comparable in the two study groups and modulated electro-hyperthermia related adverse events were limited in 8 patients presenting G1-2 skin burns.
Conclusion: The addition of mEHT to systemic CHT improved overall and progression free survival and local tumor control with comparable toxicity.
Key Words: Modulated electro-hyperthermia; Locally advanced pancreatic cancer; Tumor response; Survival
Synergy between TMZ and individualized multimodal immunotherapy to improve Overall Survival of IDH1 wild-type MGMT promoter-unmethylated GBM patients
Stefaan W. Van Gool, Jennifer Makalowski, Michael Bitar, Peter Van de Vliet, Volker Schirrmacher, Wilfried Stuecker
IOZK, Hohenstaufenring 30-32, 50674 Köln; www.iozk.de
The prognosis of IDH1 wild-type MGMT promoter-unmethylated GBM patients remains poor. Addition of Temozolomide (TMZ) to first-line local treatment shifted the median overall survival (OS) from 11.8 to 12.6 months. We retrospectively analysed the value of individualized multimodal immunotherapy (IMI) to improve OS in these patients. All adults meeting the criteria and treated 06/2015-06/2021 were selected. Thirty-two patients (12f, 20m) had a median age of 47y (range 18-69) and a KPI of 70 (50-100). Extent of resection was complete (11), <complete (12) or not documented (9). Seven patients were treated with surgery/radio(chemo)therapy and subsequent IMI (Group-1); 25 patients were treated with radiochemotherapy followed by maintenance TMZ plus IMI during and after TMZ (Group-2). Age, KPI and extent of resection were not different amongst both groups. The median OS of group-1 patients was 11m (2y OS: 0%). Surprisingly the median OS of group-2 patients was 22m with 2y OS of 36% (CI95%: 16-57), which was significantly (Log-rank: p = 0.0001) different from group-1. The data suggest that addition of IMI after local therapy on its own has no relevant effect on OS in these GBM patients, similar to maintenance TMZ. However, the combination of both TMZ + IMI significantly improved OS.
Tumors of the hepato-pancreato-biliary system: can we tame the beast?
Szász, A. Marcell, Herold, Zoltán, Garay, Tamás, Borbényi, Erika, Dank, Magdolna
Hepato-pancreatobiliary tumors are challenging to diagnose and treat, while the advanced or metastatic forms have a very low survival rate. In the past years, we have not witnessed a significant improvement in their oncological approach and outcome. A rapidly deteriorating condition is not uncommon during the course of the disease making preventing the administration of aggressive therapies. Modulated hyperthermia, with its safety profile and improved efficiency in larger tumor volumes could be a candidate for mainstream therapy in the future care of such patients. In this presentation, the rationale for utilization will be discussed based on available evidence, literature data, ongoing clinical trials and general needs of such a peculiar patient population.
In vitro and in vivo Study of the Efficacy and toxicity of combination between Modulated electro Hyperthermia and Thermo-sensitive liposomal Doxorubicin in 4T1 Triple negative breast cancer model
PhD student: Kenan Aloss, Supervisor: Dr. Peter Hamar
Translational Medicine Institute, Semmelweis University
Modulated electro-hyperthermia (mEHT) is an advanced option in the hyperthermia field, applying 13.56 Radiofrequency electromagnetic field to induce tumor-specific damage. Thermo-sensitive Liposomal Doxorubicin (TSLDOX) offers a promising approach to reduce systematic toxicity of DOX. Therefore, we hypothesized that a combination between mEHT and TSLDOX could enhance the efficacy and reduce the toxicity of Dox. Treatment efficacy will be tested by measuring tumor growth inhibition and quantifying Dox concentration in blood and tumor by spectrophotometry. In addition, body weight, hematological parameters and cardiac function will be measured to evaluate treatment toxicity. Triple-negative breast cancer cell line, 4T1, in Balb/C mice was chosen as a model for this study. A pilot experiment was done to determine the most effective and well-tolerated dose of Dox. This study showed synergism between mEHT and DOX (7.5mg/kg) as this combination significantly decreased tumor volume (P= 0.01) and tumor weight (P= 0.02) in comparison to mEHT alone. Moreover, this combined treatment had the same efficacy on tumor inhibition with less toxicity (P= 0.05) compared to Dox monotherapy with the highest dose (10mg/kg). Hematoxylin and Eosin staining demonstrated similar tumor destruction with mEHT and mEHT+ DOX without additive effect of DOX in any dose. These data suggest that the role of Dox in this synergism is limited to tumor growth inhibition. However, TSLDOX could increase tumor penetration and accumulation of Dox leading to more effect on tumor damage. Therefore, TSLDOX+ mEHT in combination will be the focus of our future work.
Non-steroidal anti-inflammatory drugs (NSAID) enhances the mEHT effect on TNBC (4T1,4T07) and melanoma(B16F10) tumor models
Nino Giunashvili, Jeremiah Thomas, Lea Danics, Csaba András Schvarcz, Pedro Viana, Kenan Aless, Mahak Zahra, Zoltán Benyó, Péter Hamar
Institute of Translational Medicine, Semmelweis University, Budapest, Hungary
Previous experiments demonstrated that 3 times Modulated Electro Hyperthermia(mEHT) treatments of 4T1 Triple Negative Breast Cancer (TNBC) mouse model caused acute phase reaction (APR), elevation of prostaglandin E2(PGE2) synthesis, optimization of the systemic immune response as well as activation of innate immunity and inhibition of tumor growth. Contrary, mEHT didn’t stimulate the adaptive immune response and secondary immunogenic cell death significantly in our TNBC (4T1, 4T07)models. Descriptive effect of mEHT could be considered as a part of the inflammation, which might have multifaceted roles in tumorigenesis, including immunosurveillance or tumor promoting effects by supporting tumor favorable microenvironment(TME). We hypothesized that inhibition of inflammation by Non-steroidal anti-inflammatory drugs (NSAID) could enhance the effect of mEHT treatment. Our group has already studied Acetylsalicylic acid (ASA) and mEHT synergistic effects on melanoma B16F10 cancer models in vivo. Thus, in our current study we investigated the synergistic effect of ASA and mEHT on 4T07 TNBC model in vivo. mEHT showed some reduction of tumor growth, although it was not significant, while combination of mEHT and ASA demonstrated significant diminish of tumor growth. TME inhibition can be one of the probable explanations of the above mentioned mEHT treatment effect, which molecular mechanism is going to be investigated further.
Modulated Electro-Hyperthermia Induces a Prominent Local Stress Response and Growth Inhibition in Triple Negative Mouse Breast Cancer
Csaba András Schvarcz 1, Lea Danics 1, Tibor Krenács 2 , Pedro Viana 1, Rita Béres 1, Tamás Vancsik 1, Ákos Nagy 3 , Zoltán Benyó 1 , Tamás Kaucsár 1 and Péter Hamar 1
1 Institute of Translational Medicine, Semmelweis University, 1094 Budapest, Hungary;
2 1st Department of Pathology and Experimental Cancer Research, Semmelweis University, 1085 Budapest, Hungary
3 Molecular Oncohematology Research Group, 1st Department of Pathology and Experimental Cancer Research, Semmelweis University, 1085 Budapest, Hungary
Introduction: Effective therapy of triple-negative breast cancer (TNBC) has not yet been achieved. Modulated electro-hyperthermia (mEHT) is a novel therapeutic option, based on the selective heating and energy transfer to the tumor tissue by an electromagnetic field.
Aims: Investigate the antitumoral effects of mEHT in TNBC mouse model and identify molecules that can serve as potential therapeutic targets.
Methods: Mouse TNBC 4T1 and 4T07 cells were inoculated orthotopically into female BALB/c mice. Tumor growth was measured by digital caliper and ultrasound (Phillips Sonos 5500). Animals were randomized into sham (n=9-10) and mEHT (n=8-11) treated groups and received treatment 3 or 5 times in every 48 hours with Labehy 200 (Oncotherm Ltd.). 24 hours after last treatment, tumors were dissected, weighed and processed. Slides were evaluated digitally (Caseviewer, 3DHistech Ltd.). Ratio of destructed vs whole tumor area (TDR%) was evaluated on H&E and cleaved caspase 3 stained slides. Heat-shock protein (HSP70) and Ki67 proliferation marker were evaluated (relative mask area %). Tumor samples were investigated with NGS and pathway analysis, and validated with Nanostring and mass spectrometry. Heat shock inhibition of KRIBB11 and effect on cell viability was studied in vitro.
Results: mEHT treatment reduced size growth (sham: 5.7x, mEHT: 2.4x relative to pre-treatment (day 6), p<0.0001) and weight (sham: 288.3±58.1 mg vs mEHT: 85.3±21.3 mg, p<0.05) in mEHT treated group, compared to sham group. HSP70 expression was 6.1x and cC3+ destructed tumor area was 6.3 x higher in treated group (p<0.05 and p<0.001) and the Ki67+ nucleus/mm2 count was significantly lower (sham: 2874 ± 193.6 pcs/mm2 vs mEHT: 1737 ± 315.3 pcs/mm2, p<0.05). NGS revealed that most upregulated genes are stress-related and fit into Response to Stimulus pathway (GO:0050896). KRIBB11 effectively decreases the expression of two identified molecules (HSP70, C4b) and tumor cell viability in vitro.
Conclusion: Repeated mEHT treatment can efficiently inhibit tumor growth and proliferation with the upregulation of several stress-related molecules. Inhibition of these molecules can potentiate mEHT’s effect and may improve the effectiveness of treatment.
Funding: NVKP_16-1-2016-0042; Dr. Korányi András az Egészségmegőrzés és Egészségkultúra Fejlesztéséért Alapítvány; EFOP-3.6.3-VEKOP-16-2017-00009; ÚNKP-21-3-II-SE-64; STIA-MEC;
139610/TMI/2020 Data of the presenter: Schvarcz Csaba András; poster presentation; Doctoral School: Doctoral School of Theoretical and Translational Medicine, Semmelweis University; e-mail: email@example.com; Supervisor: Dr. Hamar Péter
Modulated electro-hyperthermia induces angiogenic repair in murine triple negative breast cancer model
Syeda Mahak Zahra Bokhari, Lea Danics, Csaba András Schvarcz, Pedro Viana, Zoltán Benyó, Péter Hamar
Institute of Translational Medicine, Semmelweis University, Budapest, Hungary
Triple negative breast cancer (TNBC) is a highly aggressive breast cancer type. Modulated electro-hyperthermia is a novel complementary therapeutic option that can improve the effectivity of available TNBC treatments.
TNBC murine isografts were treated for three or five times with ergonomic pole electrode and LabEHY 200 device at 0.7 ± 0.3 W for 30 min every 48 hours. Tumour volume growth was monitored with ultrasound and digital calipers. Tumour destruction histology, blood capillary damage and molecular changes were detected using immunohistochemistry and RT-qPCR.
mEHT induced capillary damage in TNBC isografts, 24h after 3 mEHT treatments in a time dependent manner. 4h post treatment a significant increase in blood pools within tumour was quantified. The bleeding peaked at 12h and was significantly reduced at 24h. Similar bleeding was observed after five meht treatments and the number of viable blood vessels was significantly decreased.
Significant upregulation of stress related genes in response to mEHT treatment was observed in Multiplex analysis. An angiogenic repair in response to mEHT was detected by a reduction and subsequent continuous upregulation of tumour vasculature marker CD105. Additionally, tumour CD31 staining showed a decrease in expression at 12h followed by an upregulation after 24 hours.
Conclusively, we have strong suggestive evidence that mEHT induces blood capillary damage and triggers a stress response that repairs the tumour vasculature 24hour after the last treatment. The project will not only better elucidate the potential repair mechanisms switched on in response to mEHT but also will give information about optimal drug administration timings to observe synergistic effects of mEHT and drugs.