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4. Hibernation and Radioprotection: Gene Expression in the Liver and Testicle of Rats Irradiated under Synthetic Torpor

Author

Tinganelli, Walter, Hitrec, Timna, Romani, Fabrizio, Simoniello, Palma, Squarcio, Fabio, Stanzani, Agnese, Piscitiello, Emiliana, Marchesano, Valentina, Luppi, Marco, Sioli, Maximiliano, Helm, Alexander, Compagnone, Gaetano, Morganti, Alessio G., Amici, Roberto, Negrini, Matteo, Zoccoli, Antonio, Durante, Marco, Cerri, Matteo, Tinganelli, Walter, Hitrec, Timna, Romani, Fabrizio, Simoniello, Palma, Squarcio, Fabio, Stanzani, Agnese, Piscitiello, Emiliana, Marchesano, Valentina, Luppi, Marco, Sioli, Maximiliano, Helm, Alexander, Compagnone, Gaetano, Morganti, Alessio G., Amici, Roberto, Negrini, Matteo, Zoccoli, Antonio, Durante, Marco, and Cerri, Matteo

Abstract

Hibernation has been proposed as a tool for human space travel. In recent years, a procedure to induce a metabolic state known as "synthetic torpor" in non-hibernating mammals was successfully developed. Synthetic torpor may not only be an efficient method to spare resources and reduce psychological problems in long-term exploratory-class missions, but may also represent a countermeasure against cosmic rays. Here we show the preliminary results from an experiment in rats exposed to ionizing radiation in normothermic conditions or synthetic torpor. Animals were irradiated with 3 Gy X-rays and organs were collected 4 h after exposure. Histological analysis of liver and testicle showed a reduced toxicity in animals irradiated in torpor compared to controls irradiated at normal temperature and metabolic activity. The expression of ataxia telangiectasia mutated (ATM) in the liver was significantly downregulated in the group of animal in synthetic torpor. In the testicle, more genes involved in the DNA damage signaling were downregulated during synthetic torpor. These data show for the first time that synthetic torpor is a radioprotector in non-hibernators, similarly to natural torpor in hibernating animals. Synthetic torpor can be an effective strategy to protect humans during long term space exploration of the solar system.

Published
2024

7. FLASH with carbon ions: Tumor control, normal tissue sparing, and distal metastasis in a mouse osteosarcoma model

Author

Tinganelli, Walter, Weber, Uli, Durante, Marco, Puspitasari, Anggraeini, Simoniello, Palma, Abdollahi, Amir, Oppermann, Julius, Schuy, Christoph, Horst, Felix, Helm, Alexander, and Fournier, Claudia

Subjects
Mice, Inbred C3H, Osteosarcoma, therapy, Lung Neoplasms, Metastasis suppression, Radiotherapy Dosage, Bone Neoplasms, FLASH effect, FLASH effect,Carbon ions,Heavy ion, therapy,Mouse osteosarcoma,Metastasis suppression, Hematology, Carbon, Mice, Oncology, Animals, Female, Radiology, Nuclear Medicine and imaging, ddc:610, Protons, Carbon ions, Heavy ion, Mouse osteosarcoma
Abstract

The FLASH effect is a potential breakthrough in radiotherapy because ultra-high dose-rate irradiation can substantially widen the therapeutic window. While the normal tissue sparing at high doses and short irradiation times has been demonstrated with electrons, photons, and protons, so far evidence with heavy ions is limited to in vitro cell experiments. Here we present the first in vivo results with high-energyLM8 osteosarcoma cells were subcutaneously injected in the posterior limb of female C3H/He mice 7 days before radiation exposure. Both hind limbs of the animals were irradiated with 240 MeV/nIrradiation with carbon ions was able to control the tumour both at conventional and ultra-high dose rate. FLASH decreases normal tissue toxicity as demonstrated by the reduced structural changes in muscle compared to conventional dose-rate irradiation. Carbon ion irradiation in FLASH conditions significantly reduced lung metastasis compared to conventional dose-rate irradiation and sham-irradiated animals.We demonstrated the FLASH effect in vivo with high-energy carbon ions. In addition to normal tissue sparing, we observed tumor control and a substantial reduction of lung metastasis in an osteosarcoma mouse model.

Published
2022

8. Benzodiazepine Delorazepam Induces Locomotory Hyperactivity and Alterations in Pedal Mucus Texture in the Freshwater Gastropod Planorbarius corneus.

Author

Fogliano, Chiara, Carotenuto, Rosa, Agnisola, Claudio, Simoniello, Palma, Karam, Myriam, Manfredonia, Claudia, Avallone, Bice, and Motta, Chiara Maria

Subjects
MUCUS, SEWAGE disposal plants, FRESHWATER snails, GASTROPODA, HYPERACTIVITY, BENZODIAZEPINES, FRESH water
Abstract

Benzodiazepines, psychotropic drugs, are ubiquitous in the aquatic environment due to over-consumption and inefficient removal by sewage treatment plants. Bioaccumulation with consequent behavioral and physiological effects has been reported in many aquatic species. However, the responses are species-specific and still poorly understood. To improve the knowledge, we exposed the freshwater snail Planorbarius corneus to 1, 5, or 10 µg/L of delorazepam, the most widely consumed benzodiazepine in Italy. Conventional behavioral tests were used to assess the effects on locomotor and feeding behavior. Histological and biochemical analyses were also performed to detect possible changes in the structure and composition of the foot mucus and glands. The results show a paradoxical response with reduced feeding activity and locomotor hyperactivity. Pedal mucus was altered in texture but not in composition, becoming particularly rich in fibrous collagen-like material, and a significant change in the protein composition was highlighted in the foot. In conclusion, exposure to delorazepam induces disinhibited behavior in Planorbarius corneus, potentially increasing the risk of predation, and an increase in mucus protein production, which, together with reduced feeding activity, would severely compromise energy resources. [ABSTRACT FROM AUTHOR]

Published
2023
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14. Synthetic torpor protects rats from exposure to accelerated heavy ions

Author

Puspitasari, Anggraeini, primary, Squarcio, Fabio, additional, Quartieri, Martina, additional, Totis, Cristina, additional, Hitrec, Timna, additional, Takahashi, Akihisa, additional, Yoshida, Yukari, additional, Hanamura, Kenji, additional, Yako, Tomoko, additional, Cerri, Matteo, additional, Simoniello, Palma, additional, Durante, Marco, additional, and Tinganelli, Walter, additional

Published
2022
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16. Comparative Toxicity of Vegan Red, E124, and E120 Food Dyes on Three Rapidly Proliferating Model Systems

Author

Avallone, Bice, primary, Arena, Carmen, additional, Simoniello, Palma, additional, Di Lorenzo, Mariana, additional, Vitale, Ermenegilda, additional, Capriello, Teresa, additional, Ferrandino, Ida, additional, Raggio, Anja, additional, Sasso, Margherita, additional, Napolitano, Gaetana, additional, De Bonis, Salvatore, additional, Fogliano, Chiara, additional, Agnisola, Claudio, additional, and Motta, Chiara Maria, additional

Published
2022
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17. Biological Cardiac Tissue Effects of High-Energy Heavy Ions – Investigation for Myocardial Ablation

Author

Rapp, Felicitas, Simoniello, Palma, Wiedemann, Julia, Bahrami, Karola, Grünebaum, Valeria, Ktitareva, Svetlana, Durante, Marco, Lugenbiel, Patrick, Thomas, Dierk, Lehmann, Helge Immo, Packer, Douglas L., Graeff, Christian, Fournier, Claudia, Rapp, Felicitas, Simoniello, Palma, Wiedemann, Julia, Bahrami, Karola, Grünebaum, Valeria, Ktitareva, Svetlana, Durante, Marco, Lugenbiel, Patrick, Thomas, Dierk, Lehmann, Helge Immo, Packer, Douglas L., Graeff, Christian, and Fournier, Claudia

Abstract

Noninvasive X-ray stereotactic treatment is considered a promising alternative to catheter ablation in patients affected by severe heart arrhythmia. High-energy heavy ions can deliver high radiation doses in small targets with reduced damage to the normal tissue compared to conventional X-rays. For this reason, charged particle therapy, widely used in oncology, can be a powerful tool for radiosurgery in cardiac diseases. We have recently performed a feasibility study in a swine model using high doses of high-energy C-ions to target specific cardiac structures. Interruption of cardiac conduction was observed in some animals. Here we report the biological effects measured in the pig heart tissue of the same animals six months after the treatment. Immunohistological analysis of the target tissue showed (1.) long-lasting vascular damage, i.e. persistent hemorrhage, loss of microvessels, and occurrence of siderophages, (2.) fibrosis and (3.) loss of polarity of targeted cardiomyocytes and wavy fibers with vacuolization. We conclude that the observed physiological changes in heart function are produced by radiation-induced fibrosis and cardiomyocyte functional inactivation. No effects were observed in the normal tissue traversed by the particle beam, suggesting that charged particles have the potential to produce ablation of specific heart targets with minimal side effects.

Published
2022

18. Commercial Red Food Dyes Preparations Modulate the Oxidative State in Three Model Organisms (Cucumis sativus, Artemia salina, and Danio rerio)

Author

Napolitano, Gaetana, primary, Motta, Chiara Maria, additional, Agnisola, Claudio, additional, Venditti, Paola, additional, Fasciolo, Gianluca, additional, Ferrandino, Ida, additional, Capriello, Teresa, additional, Vitale, Ermenegilda, additional, Costanzo, Giulia, additional, Avallone, Bice, additional, Simoniello, Palma, additional, and Arena, Carmen, additional

Published
2022
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26. In Reply to Elmali et al

Author

Helm, Alexander, primary, Tinganelli, Walter, additional, Fournier, Claudia, additional, Simoniello, Palma, additional, Kurosawa, Fuki, additional, Shimokawa, Takashi, additional, and Durante, Marco, additional

Published
2021
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29. 'IMMUNOGENIC POTENTIAL OF CARBON IONS IN COMBINATION WITH IMMUNOTHERAPEUTIC DRUGS TESTED IN A MOUSE MODEL'

Author

Simoniello, Palma, Alexander, Helm, Walter, Tinganelli, Fuki, Kurosawa, Claudia, Fournier, and Takashi Shimokawa and Marco, Durante.

Subjects
Radiotherapy is one of the main treatments for cancers, in turn resulting in the shrinkage of out-of-field metastases. This phenomenon is known as abscopal effect. The right combination of radiotherapy with immunotherapy could be a strategy to induce durable responses and improve survival. Particle therapy with protons and especially carbon-ions is assumed to further increase the immunogenic effect. The aim of this study was to investigate the efficacy of X-ray and high-energy carbon ion therapy in a combination with immunotherapy. In this work we investigated the combination of radiotherapy, i.e. it may induce an immunogenic cell death in tumor cells leading to immunoactivation, LM8 cells). Tumor cells were injected in both hind limbs of the mice. Following an initial growth period, metastatic disease still remains a problem. It is known that immune system plays a key role in the elimination of primary tumors and preventing tumor recurrence. Ionizing radiation is reported to be immunogenic, Radiotherapy is one of the main treatments for cancers, which has experienced important improvements in the last years, metastatic disease still remains a problem. It is known that immune system plays a key role in the elimination of primary tumors and preventing tumor recurrence. Ionizing radiation is reported to be immunogenic, i.e. it may induce an immunogenic cell death in tumor cells leading to immunoactivation, in turn resulting in the shrinkage of out-of-field metastases. This phenomenon is known as abscopal effect. The right combination of radiotherapy with immunotherapy could be a strategy to induce durable responses and improve survival. Particle therapy with protons and especially carbon-ions is assumed to further increase the immunogenic effect. The aim of this study was to investigate the efficacy of X-ray and high-energy carbon ion therapy in a combination with immunotherapy. In this work we investigated the combination of radiotherapy, with either carbon ions or X-rays, and immune checkpoint inhibitors (CPI) (anti- CTLA-4 and anti-PD-1), in a syngeneic osteosarcoma mouse model (C3H/He mice, LM8 cells). Tumor cells were injected in both hind limbs of the mice. Following an initial growth period, one of the tumors was irradiated while the other was kept out of the irradiation field, mimicking the abscopal tumor. Macroscopic analysis on out-of-field metastases were carried out to investigate spontaneously formed lung metastases. Immune responses were investigated by histological analyses on the abscopal tumors screening. The results show that combination of carbon ion therapy and CPI lead to a reduced growth of abscopal tumors and reduced formation of lung metastases. In addition, in the abscopal tumors treated with combining therapy an immunogenic response resulting in increasing of infiltration of CD8+ cells was observed. Combination of CPI with high-energy carbon ion radiotherapy could be a promising strategy for the treatment of tumor and metastasis, in a syngeneic osteosarcoma mouse model (C3H/He mice, one of the tumors was irradiated while the other was kept out of the irradiation field, and immune checkpoint inhibitors (CPI) (anti- CTLA-4 and anti-PD-1), with either carbon ions or X-rays, which has experienced important improvements in the last years, mimicking the abscopal tumor. Macroscopic analysis on out-of-field metastases were carried out to investigate spontaneously formed lung metastases. Immune responses were investigated by histological analyses on the abscopal tumors screening. The results show that combination of carbon ion therapy and CPI lead to a reduced growth of abscopal tumors and reduced formation of lung metastases. In addition, in the abscopal tumors treated with combining therapy an immunogenic response resulting in increasing of infiltration of CD8+ cells was observed. Combination of CPI with high-energy carbon ion radiotherapy could be a promising strategy for the treatment of tumor and metastasis
Published
2020

31. Gene expression in the liver of the rat induced in synthetic torpor

Author

Hitrec, Timna, primary, Tinganelli, Walter, additional, Romani, Fabrizio, additional, Simoniello, Palma, additional, Squarcio, Fabio, additional, Piscitiello, Emiliana, additional, Marchesano, Valentina, additional, Luppi, Marco, additional, Occhinegro, Alessandra, additional, Sioli, Maximiliano, additional, Helm, Alexander, additional, Compagnone, Gaetano, additional, Morganti, Alessio G, additional, Amici, Roberto, additional, Negrini, Matteo, additional, Zoccoli, Antonio, additional, Durante, Marco, additional, and Cerri, Matteo, additional

Published
2020
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33. Biological Cardiac Tissue Effects of High-Energy Heavy Ions – Investigation for Myocardial Ablation

Author

Simoniello, Palma, Wiedemann, Julia, Bahrami, Karola, Grünebaum, Valeria, Ktitareva, Svetlana, Durante, Marco, Lugenbiel, P., Thomas, D., Lehmann, H. Immo, Packer, Douglas L., and Graeff, Christian

Abstract

Scientific reports 9(1), 5000 (2019). doi:10.1038/s41598-019-41314-x, Published by Macmillan Publishers Limited, part of Springer Nature, [London]

Published
2019
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34. Combination immunotherapy-radiation therapy using particles with Ipilimumab and Nivolumab in mouse models

Author

Helm, Alexander, Simoniello, Palma, Bisio, Alessandra, Marchesano, Valentina, Durante, Marco, Tinganelli, Walter, Kurosawa, Fuki, Ebner, Daniel, and Shimokawa, Takashi

Abstract

Background Combination radiotherapy and immunotherapy shows highly promising clinical results and is therefore a regimen of growing interest. Ionizing radiation is reported to be immunogenic, i.e. it may induce an immunogenic cell death in tumor cells leading to immunoactivation, in turn resulting in the shrinkage of out-of-field metastases. This phenomenon is termed the abscopal effect. Particle therapy with protons and especially carbon-ions is assumed to further increase the immunogenic effect. Brief methods We set up a combination of particle radiation (carbon ions or protons) together with Ipilimumab and/or Nivolumab before or after exposure, so as to enhance the abscopal effect in mouse models (C3H/He and C57BL/6) harboring tumors originating from B16 melanoma cells and LM8 osteosarcoma, respectively. Results In a C57BL/6-B16 model, a combination of carbon ion therapy and Ipilimumab/Nivolumab tended to result in a reduced growth of abscopal tumors in comparison with carbon ions or Ipilimumab/Nivolumab only. In a C3H/He-LM8 model, the combination of carbon ion therapy and Ipilimumab/Nivolumab resulted in a reduced growth of the abscopal tumor and reduced formation of lung metastases. Conclusions The model seems suitable to study the optimal temporally application of drugs in combination with radiation. Further experiments are currently under analysis. Here we present our group's first results., the 3th international conference of Immunotherapy Radiotherapy combinations

Published
2018

36. PROTECTION FROM X-RAY DAMAGE IN RATS INDUCED INTO SYNTHETIC TORPOR

Author

Simoniello, Palma, Timna, Hitrec, Walter, Tinganelli, Matteo, Cerri, Fabio, Squarcio, Roberto, Amici, Emiliana, Piscitiello, Marco, Luppi, Massimiliano, Sioli, Alexander, Helm, Alessio, Morganti, Matteo, Negrini, and Marco Durante and Antonio Zoccoli

Published
2018

38. Effects of four food dyes on development of three model species, Cucumis sativus, Artemia salina and Danio rerio: Assessment of potential risk for the environment

Author

Motta, Chiara Maria, primary, Simoniello, Palma, additional, Arena, Carmen, additional, Capriello, Teresa, additional, Panzuto, Raffaele, additional, Vitale, Ermenegilda, additional, Agnisola, Claudio, additional, Tizzano, Monica, additional, Avallone, Bice, additional, and Ferrandino, Ida, additional

Published
2019
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40. Hibernation and Radioprotection: Gene Expression in the Liver and Testicle of Rats Irradiated under Synthetic Torpor

Author

Tinganelli, Walter, primary, Hitrec, Timna, additional, Romani, Fabrizio, additional, Simoniello, Palma, additional, Squarcio, Fabio, additional, Stanzani, Agnese, additional, Piscitiello, Emiliana, additional, Marchesano, Valentina, additional, Luppi, Marco, additional, Sioli, Maximiliano, additional, Helm, Alexander, additional, Compagnone, Gaetano, additional, Morganti, Alessio, additional, Amici, Roberto, additional, Negrini, Matteo, additional, Zoccoli, Antonio, additional, Durante, Marco, additional, and Cerri, Matteo, additional

Published
2019
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41. Exposure to low doses of p,p’-DDE exerts testicular toxicity in male rats

Author

MIGLIACCIO, VINCENZO, SIMONIELLO, PALMA, PUTTI, ROSALBA, LIONETTI, LILLA', SCUDIERO, ROSARIA, Sica, Raffaela, Universitat de Barcelona, Migliaccio, Vincenzo, Sica, Raffaela, Simoniello, Palma, Putti, Rosalba, Lionetti, Lilla', and Scudiero, Rosaria

Subjects
DDE toxicity, testis, rattus
Published
2016

43. Generating and grading the abscopal effect: Proposal for comprehensive evaluation of combination immunoradiotherapy in mouse models

Author

Ebner, Daniel K., Tinganelli, Walter, Helm, Alexander, Bisio, Alessandra, Simoniello, Palma, Natale, Francesco, Yamada, Shigeru, Kamada, Tadashi, Shimokawa, Takashi, Durante, Marco, Collaboration, the Abscopal Research, Ebner, D. K., Tinganelli, W., Helm, A., Bisio, A., Simoniello, P., Natale, F., Yamada, S., Kamada, T., Shimokawa, T., and Durante, M.

Subjects
Cancer Research, Oncology, Particle therapy, Radiology, Nuclear Medicine and imaging, Carbon-ion radiotherapy, Immunotherapy
Abstract

Numerous published case reports and forthcoming clinical trials combine immunotherapy and radiotherapy with the goal of demonstrating an abscopal effect reaction. However, reports and results are varied, and a comprehensive animal study combining radiotherapy modalities and immunotherapy agents has yet to be performed. Further, clinical reports are mixed, with inconsistent generation of the effect as well as abscopal effects seen ranging from widespread disease elimination to growth stagnation of local lymph node metastasis. We propose a grading system for use in differentiating abscopal effects seen in animal and clinical trials. Further, we will conduct a comprehensive study in mice, evaluating three radiotherapeutic modalities (photon, proton, and carbon-ion) combined with five immunotherapeutic agents with differing actions. Comprehensive cell analysis will be conducted with the aim of improving upon draft models of abscopal effect generation, as well as differentiating between locoregional and systemic methods of action. Further, evaluation of radiation fractionation, as well as combination with ex vivo activated dendritic cell (DC) inoculation, will be performed. Though the abscopal effect has been reported for nearly 70 years, only recently has delineation of its mechanism seemed possible. Comprehensive evaluation of available modalities may shed light on the precise requirements for generating the effect, potentially enabling its regular usage in the treatment of disease.

Published
2017

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