Determining the effects of hyperthermia on the tumor and acute normal tissue response of FLASH radiation

Authors

DOI:

https://doi.org/10.2340/1651-226X.2025.44043

Keywords:

Electron radiation, Conventional dose rate (CONV), Ultra-high dose rate (FLASH), Hyperthermia, Tumor growth time, Acute skin toxicity

Abstract

Introduction: There is limited indication of how hyperthermia would influence the unique proposition of FLASH radiation – its ability to maintain comparable tumor response while offering protection to normal tissues. Hence, this study was designed to investigate the potential of combining FLASH radiation with hyperthermia.

Materials and methods: Experiments were performed using female CDF1 mice, where the tumor bearing or non-tumor bearing right hind legs were irradiated with either conventional dose rate (CONV) or FLASH radiation ± hyperthermia. Hyperthermia was applied 30 minutes after radiation at 42.5°C for 60 minutes. The tumor endpoint was growth delay to three times its initial treatment volume (TGT3) and the normal tissue endpoint was an acute skin toxicity of score 2.5 and above, characterized by moderate moist desquamation and partial leg deformity.

Results: In tumor studies, the thermal enhancement ratio (TER) was 1.68 for FLASH radiotherapy and 1.50 for conventional (CONV) radiation. In acute skin toxicity studies, the TER was slightly lower, at 1.37 for FLASH and 1.29 for CONV. The dose modifying factor (DMF) in tumor studies was 1.12 but decreased to 1.00 when hyperthermia was added. Similarly, in acute skin toxicity studies, the DMF was initially 1.53 and dropped to 1.45 with the addition of hyperthermia.

Interpretation: Hyperthermia significantly sensitized both the CONV and FLASH radiation, but the enhancement is comparable between the two different dose rate radiations in both tumors and normal tissues.

Downloads

Download data is not yet available.

References

Favaudon V, Caplier L, Monceau V, Pouzoulet F, Sayarath M, Fouillade C, et al. Ultrahigh dose-rate FLASH irradiation increases the differential response between normal and tumor tissue in mice. Sci Transl Med. 2014;6:245ra93.

https://doi.org/10.1126/scitranslmed.3008973 DOI: https://doi.org/10.1126/scitranslmed.3008973

Böhlen TT, Germond J-F, Petersson K, Ozsahin EM, Herrera FG, Bailat C, et al. Effect of conventional and ultrahigh dose rate FLASH irradiations on preclinical tumor models: a systematic analysis. Int J Radiat Oncol Biol Phys. 2023;117:1007–17.

https://doi.org/10.1016/j.ijrobp.2023.05.045 DOI: https://doi.org/10.1016/j.ijrobp.2023.05.045

Böhlen TT, Germond J-F, Bourhis J, Vozenin M-C, Ozsahin EM, Bochud F, et al. Normal tissue sparing by FLASH as a function of single-fraction dose: a quantitative analysis. Int J Radiat Oncol Biol Phys. 2022;114:1032–44.

https://doi.org/10.1016/j.ijrobp.2022.05.038 DOI: https://doi.org/10.1016/j.ijrobp.2022.05.038

Sørensen BS, Sitarz MK, Ankjærgaard C, Johansen JG, Andersen CE, Kanouta E, et al. Pencil beam scanning proton FLASH maintains tumor control while normal tissue damage is reduced in a mouse model. Radiother Oncol. 2022;175:178–84.

https://doi.org/10.1016/j.radonc.2022.05.014 DOI: https://doi.org/10.1016/j.radonc.2022.05.014

Cunningham S, McCauley S, Vairamani K, Speth J, Girdhani S, Abel E, et al. FLASH proton pencil beam scanning irradiation minimizes radiation-induced leg contracture and skin toxicity in mice. Cancers (Basel) 2021;13:1012.

https://doi.org/10.3390/cancers13051012 DOI: https://doi.org/10.3390/cancers13051012

Lin B, Huang D, Gao F, Yang Y, Wu D, Zhang Y, et al. Mechanisms of FLASH effect. Front Oncol. 2022;12:995612.

https://doi.org/10.3389/fonc.2022.995612 DOI: https://doi.org/10.3389/fonc.2022.995612

Overgaard J. The current and potential role of hyperthermia in radiotherapy. Int J Radiat Oncol Biol Phys. 1989;16:535–49.

https://doi.org/10.1016/0360-3016(89)90470-7 DOI: https://doi.org/10.1016/0360-3016(89)90470-7

Elming PB, Sørensen BS, Oei AL, Franken NAP, Crezee J, Overgaard J, et al. Hyperthermia: the optimal treatment to overcome radiation resistant hypoxia. Cancers (Basel). 2019;11:60.

https://doi.org/10.3390/cancers11010060 DOI: https://doi.org/10.3390/cancers11010060

Oei AL, Kok HP, Oei SB, Horsman MR, Stalpers LJA, Franken NAP, et al. Molecular and biological rationale of hyperthermia as radio- and chemosensitizer. Adv Drug Deliv Rev. 2020;163–164:84–97.

https://doi.org/10.1016/j.addr.2020.01.003 DOI: https://doi.org/10.1016/j.addr.2020.01.003

Dewey WC. Interaction of heat with radiation and chemotherapy. Cancer Res. 1984;44:4714–20.

Horsman MR, Overgaard J. Hyperthermia: a potent enhancer of radiotherapy. Clin Oncol (R Coll Radiol). 2007;19:418–26.

https://doi.org/10.1016/j.clon.2007.03.015 DOI: https://doi.org/10.1016/j.clon.2007.03.015

Datta NR, Ordóñez SG, Gaipl US, Paulides MM, Crezee H, Gellermann J, et al. Local hyperthermia combined with radiotherapy and-/or chemotherapy: recent advances and promises for the future. Cancer Treat Rev. 2015;41:742–53.

https://doi.org/10.1016/j.ctrv.2015.05.009 DOI: https://doi.org/10.1016/j.ctrv.2015.05.009

Peeken JC, Vaupel P, Combs SE. Integrating hyperthermia into modern radiation oncology: what evidence is necessary? Front Oncol. 2017;7:132.

https://doi.org/10.3389/fonc.2017.00132 DOI: https://doi.org/10.3389/fonc.2017.00132

Wang Z, Armour EP, Corry PM, Martinez A. Elimination of dose-rate effects by mild hyperthermia. Int J Radiat Oncol Biol Phys. 1992;24:965–73.

https://doi.org/10.1016/0360-3016(92)90481-v DOI: https://doi.org/10.1016/0360-3016(92)90481-V

Sakurai H, Mitsuhasi N, Takahashi T, Hashida I, Niibe H. Enhanced cytotoxicity in combination of low dose-rate irradiation with hyperthermia in vitro. Int J Hyperthermia. 1996;12:355–66.

https://doi.org/10.3109/02656739609022524 DOI: https://doi.org/10.3109/02656739609022524

Armour EP, Wang Z, Corry PM, Chen PY, Martinez A. Hyperthermic enhancement of high dose-rate irradiation in 9L gliosarcoma cells. Int J Radiat Oncol Biol Phys. 1994;28:171–7.

https://doi.org/10.1016/0360-3016(94)90155-4 DOI: https://doi.org/10.1016/0360-3016(94)90155-4

Krossnes BK, Schem B-C, Nygaard B, Dahl O, Mella O. Interaction between microwave-induced brain hyperthermia and high dose rate radiation in the BT4An brain glioma in rats. J Neurooncol. 1996;27:205–14.

https://doi.org/10.1007/BF00165476 DOI: https://doi.org/10.1007/BF00165476

Suo M, Shen H, Lyu M, Jiang Y, Liao X, Tang W, et al. Biomimetic nano‐cancer stem cell scavenger for inhibition of breast cancer recurrence and metastasis after FLASH‐radiotherapy. Small. 2024;20:e2400666.

https://doi.org/10.1002/smll.202400666 DOI: https://doi.org/10.1002/smll.202400666

Shen H, Wang H, Mo J, Zhang J, Xu C, Sun F, et al. Unrestricted molecular motions enable mild photothermy for recurrence-resistant FLASH antitumor radiotherapy. Bioact Mater. 2024;37:299–312.

https://doi.org/10.1016/j.bioactmat.2024.03.024 DOI: https://doi.org/10.1016/j.bioactmat.2024.03.024

Præstegaard LH. Radiation safety of ultra-high dose rate electron accelerators for FLASH radiotherapy. Med Phys. 2024;51:6206–19.

https://doi.org/10.1002/mp.17245 DOI: https://doi.org/10.1002/mp.17245

Overgaard CB, Reaz F, Sitarz M, Poulsen P, Overgaard J, Bassler N, et al. An experimental setup for proton irradiation of a murine leg model for radiobiological studies. Acta Oncol (Madr). 2023;62:1566–73.

https://doi.org/10.1080/0284186X.2023.2246641 DOI: https://doi.org/10.1080/0284186X.2023.2246641

Kristensen L, Poulsen PR, Kanouta E, Rohrer S, Ankjærgaard C, Andersen CE, et al. Spread-out Bragg peak FLASH: quantifying normal tissue toxicity in a murine model. Front Oncol. 2024;14:1427667.

https://doi.org/10.3389/fonc.2024.1427667 DOI: https://doi.org/10.3389/fonc.2024.1427667

Sørensen BS, Bassler N, Nielsen S, Horsman MR, Grzanka L, Spejlborg H, et al. Relative biological effectiveness (RBE) and distal edge effects of proton radiation on early damage in vivo. Acta Oncol. 2017;56:1387–91.

https://doi.org/10.1080/0284186X.2017.1351621 DOI: https://doi.org/10.1080/0284186X.2017.1351621

Sørensen BS, Krzysztof Sitarz M, Ankjærgaard C, Johansen J, Andersen CE, Kanouta E, et al. In vivo validation and tissue sparing factor for acute damage of pencil beam scanning proton FLASH. Radiother Oncol. 2022;167:109–15.

https://doi.org/10.1016/j.radonc.2021.12.022 DOI: https://doi.org/10.1016/j.radonc.2021.12.022

Kristensen L, Rohrer S, Hoffmann L, Præstegaard LH, Ankjærgaard C, Andersen CE, et al. Electron vs proton FLASH radiation on murine skin toxicity. Radiother Oncol. 2025;206:110796.

https://doi.org/10.1016/j.radonc.2025.110796 DOI: https://doi.org/10.1016/j.radonc.2025.110796

Lindegaard JC, Overgaard J. Factors of importance for the development of the step-down heating effect in a C3H mammary

carcinoma in vivo. Int J Hyperthermia. 1987;3:79–91.

https://doi.org/10.3109/02656738709140375 DOI: https://doi.org/10.3109/02656738709140375

Horsman MR, Christensen KL, Overgaard J. Hydralazine-induced enhancement of hyperthermic damage in a C3H mammary carcinoma in vivo. Int J Hyperthermia. 1989;5:123–36.

https://doi.org/10.3109/02656738909140442 DOI: https://doi.org/10.3109/02656738909140442

von der Maase H. Effect of cancer chemotherapeutic drugs on the radiation-induced skin reactions in mouse feet. Br J Radiol. 1984;57:697–707.

https://doi.org/10.1259/0007-1285-57-680-697 DOI: https://doi.org/10.1259/0007-1285-57-680-697

GraphPad Software. GraphPad Prism for macOS, version 10.3.1. San Diego (CA): Dotmatics; 2024. Available from: https://www.graphpad.com

Sørensen BS, Kanouta E, Ankjærgaard C, Kristensen L, Johansen JG, Sitarz MK, et al. Proton FLASH: impact of dose rate and split dose on acute skin toxicity in a murine model. Int J Radiat Oncol Biol Phys. 2024;120:265–75.

https://doi.org/10.1016/j.ijrobp.2024.04.071 DOI: https://doi.org/10.1016/j.ijrobp.2024.04.071

Saladino C, Ben-Hur E. Heat-enhanced radio response in HeLa cells. Isr J Med Sci. 1976;12:134–8.

Baker DG, Sager HT, Constable WC. The response of a solid tumor to X-irradiation as modified by dose rate, fractionation, and hyperthermia. Cancer Invest. 1987;5:409–16.

https://doi.org/10.3109/07357908709032898 DOI: https://doi.org/10.3109/07357908709032898

Gerner EW, Oval JH, Manning MR, Sim DA, Bowden GT, Hevezi JM. Dose-rate dependence of heat radiosensitization. Int J Radiat Oncol Biol Phys. 1983;9:1401–4.

https://doi.org/10.1016/0360-3016(83)90274-2 DOI: https://doi.org/10.1016/0360-3016(83)90274-2

Pottern CS. Radiation effects on keratoblasts. Radiation Skin. London: Taylor & Francis; 1985, p. 102–52.

Leavitt RJ, Almeida A, Grilj V, Montay-Gruel P, Godfroid C, Petit B, et al. Acute hypoxia does not alter tumor sensitivity to FLASH radiation therapy. Int J Radiat Oncol Biol Phys. 2024;119:1493–505.

https://doi.org/10.1016/j.ijrobp.2024.02.015 DOI: https://doi.org/10.1016/j.ijrobp.2024.02.015

Overgaard J. Effect of hyperthermia on the hypoxic fraction in an experimental mammary carcinoma in vivo. Br J Radiol. 1981;54:245–9.

https://doi.org/10.1259/0007-1285-54-639-245 DOI: https://doi.org/10.1259/0007-1285-54-639-245

Published

2025-10-21

How to Cite

Sinha, P. M., Kristensen, L., Folefac, C. A., Præstegaard, L. H., Hoffmann, L., Poulsen, P. R., … Sørensen, B. S. (2025). Determining the effects of hyperthermia on the tumor and acute normal tissue response of FLASH radiation. Acta Oncologica, 64, 1437–1445. https://doi.org/10.2340/1651-226X.2025.44043