2012 Activity Report of the Regional Research Programme on Hadrontherapy for the ETOILE Center

2012 is the penultimate year of financial support by the CPER 2007-2013 for ETOILE's research program, sustained by the PRRH at the University Claude Bernard. As with each edition we make the annual review of the research in this group, so active for over 12 years now. Over the difficulties in the decision-making process for the implementation of the ETOILE Center, towards which all our efforts are focussed, some "themes" (work packages) were strengthened, others have progressed, or have been dropped. This is the case of the eighth theme (technological developments), centered around the technology for rotative beam distribution heads (gantries) and, after being synchronized with the developments of ULICE's WP6, remained so by ceasing its activities, coinciding also with the retirement of its historic leader at IPNL, Marcel Bajard. Topic number 5 ("In silico simulations") has suffered the departure of its leader, Benjamin Ribba, although the work has still been provided by Branka Bernard, a former postdoctoral fellow in Lyon Sud, and now back home in Croatia, still in contract with UCBL for the ULICE project. Aside from these two issues (and the fact that the theme "Medico-economical simulations" is now directly linked to the first one ("Medical Project"), the rest of the teams are growing, as evidenced by the publication statistics at the beginning of this report. This is obviously due to the financial support of our always faithful regional institutions, but also to the synergy that the previous years, the European projects, the arrival of the PRIMES LabEx, and the national France Hadron infrastructure have managed to impulse. The Rhone-Alpes hadron team, which naturally includes the researchers of LPC at Clermont, should also see its influence result in a strong presence in France Hadron's regional node, which is being organized. The future of this regional research is not yet fully guaranteed, especially in the still uncertain context of ETOILE, but the tracks are beginning to emerge to allow past and present efforts translate into a long future that we all want to see established. Each of the researchers in PRRH is aware that 2013 will be (and already is) the year of great challenge : for ETOILE, for the PRRH, for hadron therapy in France, for French hadrontherapy in Europe (after the opening and beginning of treatments in the German [HIT Heidelberg, Marburg], Italian [CNAO, Pavia] and Austrian [MedAustron, Wien Neuerstadt]) centers. Let us meet again in early 2014 for a comprehensive review of the past and a perspective for the future ..

Transient Alteration of Cellular Redox Buffering before Irradiation Triggers Apoptosis in Head and Neck Carcinoma Stem and Non-Stem Cells

Background: Head and neck squamous cell carcinoma (HNSCC) is an aggressive and recurrent malignancy owing to intrinsic radioresistance and lack of induction of apoptosis. The major focus of this work was to design a transient glutathione depleting strategy during the course of irradiation of HNSCC in order to overcome their radioresistance associated with redox adaptation. Methodology/Principal Findings: Treatment of SQ20B cells with dimethylfumarate (DMF), a GSH-depleting agent, and L-Buthionine sulfoximine (BSO), an inhibitor of GSH biosynthesis 4 h before a 10 Gy irradiation led to the lowering of the endogenous GSH content to less than 10 % of that in control cells and to the triggering of radiation-induced apoptotic cell death. The sequence of biochemical events after GSH depletion and irradiation included ASK-1 followed by JNK activation which resulted in the triggering of the intrinsic apoptotic pathway through Bax translocation to mitochondria. Conclusions: This transient GSH depletion also triggered radiation-induced cell death in SQ20B stem cells, a key event to overcome locoregional recurrence of HNSCC. Finally, our in vivo data highlight the relevance for further clinical trials o

Transient Glutathione Depletion before Irradiation triggers Apoptosis in Head and Neck Squamous Cell Carcinoma

Glutathione, a major intra-cellular antioxidant, has been reported to play a fundamental role in the resistance of some cancer cells to radiotherapy so that this molecule can be considered as a potential clinical target for their radio-sensitization. The radio-resistant head and neck squamous carcinomas cell line SQ20B was shown to display a high endogenous level of reduced glutathione, up to 8-fold when compared to its radiosensitive counterpart SCC61 cell line. This result led us to consider the endogenous reduced glutathione as a key factor involved in the resistance of SQ20B cells to ionizing radiation. The purpose of this study was to experiment the incidence of a short and transient depletion of intracellular glutathione before irradiation of SQ20B cells. We therefore made use of the combination of dimethylfumarate (DMF), a glutathione-depleting agent in association with buthionine sulfoximine (BSO), a specific inhibitor of glutamate-cysteine ligase, the first enzyme of the glutathione biosynthesis pathway. In preliminary experiments, we have determined both the efficiency in lowering the endogenous glutathione level and the potential cyto-toxicity of these molecules in un-irradiated SQ20B cells. Although no toxicity was evidenced, a 95 % decrease of intracellular glutathione was obtained under our experimental conditions. This treatment combined to irradiation led to the triggering of apoptosis of the radioresistant SQ20B cell line which significantly increased from 48 hours to longer times, as evidenced by an enhancement of 50 % of the activity of caspases and of the number of cells in the sub-G1 phase, up to 55 % 96h after irradiation. Triggering of apoptosis in SQ210B cells was found to involve the c-Jun N-terminal Kinase (JNK) pathway which was phosphorylated 2 h after irradiation. This activation of JNK was dependent upon the generation of radical oxygen species which resulted in the dissociation of thioredoxin-ASK 1 complex (MAPKKK of JNK pathway). The phosphorylated form of JNK led to the activation and translocation of Bax to mitochondria which further resulted in the alteration of the organelle, as evidenced by the loss of the mitochondrial transmembrane potential (ΔΨm) and the increase of the secondary radical species generated through the mitochondrial respiratory chain. Taken altogether, our results demonstrate that a transient glutathione depletion before irradiation can trigger apoptosis of radio-resistant SQ20B cells through the activation of a JNK-dependent pathway. Moreover, the absence of a significant toxicity of this pharmacological treatment is of fundamental importance for further in vivo studies

Gadolinium-based nanoparticles can overcome the radioresistance of head and neck squamous cell carcinoma through the induction of autophagy

International audienceRadiation therapy is a mainstay in the therapeutic management of Head and Neck Squamous Cell Carcinoma (HNSCC). Despite significant progress in this field, radioresistance still accounts for most treatment failures. Gadolinium-based nanoparticles (GBNs) have shown great promises as radiosensitizers but the underlying sensitizing mechanism is still largely unknown with regards to the disparities obtained in in vitro studies. In this study, we show that a new formulation of GBNs, AGuIX®, can radiosensitize HNSCC after cell uptake and further accumulation in lysosomes. Although radiation alone triggered late apoptosis and mitochondrial impairment, the pre-treatment with GBNs led to complex DNA damage and a specific increase of autophagic cell death. In addition, a significant radio-enhancement effect was obtained after the pre-conditioning of cells with a glutathione inhibitor before GBNs treatment and radiation exposure. Overall, our results provide additional information on the radio-enhancing properties of GBNs in the management of radioresistant HNSCC

Flat band and Lifshitz transition in long-range-ordered supergraphene obtained by Erbium intercalation

International audienceWe report the observation of graphene doped up to the Lifshitz transition obtained solely by the intercalation of Erbium atoms. ARPES measurements show that a wide flat band is generated around the M point of the Brillouin zone. We propose that this is the effect of an induced spin-orbit coupling. Scanning Tunneling Microscopy (STM) experiments reveal a longrange ordered hexagonal superstructure: we argue that this reflects the ordering of the intercalated Er atoms acting as impurities. This system provides a playground to study the interaction between a magnetic order and a divergent density of states at the Fermi level

2012 Activity Report of the Regional Research Programme on Hadrontherapy for the ETOILE Center

2012 is the penultimate year of financial support by the CPER 2007-2013 for ETOILE's research program, sustained by the PRRH at the University Claude Bernard. As with each edition we make the annual review of the research in this group, so active for over 12 years now. Over the difficulties in the decision-making process for the implementation of the ETOILE Center, towards which all our efforts are focussed, some "themes" (work packages) were strengthened, others have progressed, or have been dropped. This is the case of the eighth theme (technological developments), centered around the technology for rotative beam distribution heads (gantries) and, after being synchronized with the developments of ULICE's WP6, remained so by ceasing its activities, coinciding also with the retirement of its historic leader at IPNL, Marcel Bajard. Topic number 5 ("In silico simulations") has suffered the departure of its leader, Benjamin Ribba, although the work has still been provided by Branka Bernard, a former postdoctoral fellow in Lyon Sud, and now back home in Croatia, still in contract with UCBL for the ULICE project. Aside from these two issues (and the fact that the theme "Medico-economical simulations" is now directly linked to the first one ("Medical Project"), the rest of the teams are growing, as evidenced by the publication statistics at the beginning of this report. This is obviously due to the financial support of our always faithful regional institutions, but also to the synergy that the previous years, the European projects, the arrival of the PRIMES LabEx, and the national France Hadron infrastructure have managed to impulse. The Rhone-Alpes hadron team, which naturally includes the researchers of LPC at Clermont, should also see its influence result in a strong presence in France Hadron's regional node, which is being organized. The future of this regional research is not yet fully guaranteed, especially in the still uncertain context of ETOILE, but the tracks are beginning to emerge to allow past and present efforts translate into a long future that we all want to see established. Each of the researchers in PRRH is aware that 2013 will be (and already is) the year of great challenge : for ETOILE, for the PRRH, for hadron therapy in France, for French hadrontherapy in Europe (after the opening and beginning of treatments in the German [HIT Heidelberg, Marburg], Italian [CNAO, Pavia] and Austrian [MedAustron, Wien Neuerstadt]) centers. Let us meet again in early 2014 for a comprehensive review of the past and a perspective for the future ..