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A New Application of Microbeam Radiation Therapy (MRT) on the Treatment of Epilepsy and Brain Disorders

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A New Application of Microbeam Radiation Therapy (MRT) on the Treatment of Epilepsy and Brain Disorders A new application of microbeam radiation therapy (MRT) on the treatment of epilepsy and brain disorders. Erminia Fardone To cite this version: Erminia Fardone. A new application of microbeam radiation therapy (MRT) on the treatment of epilepsy and brain disorders.. Human health and pathology. Université de Grenoble, 2013. English. NNT : 2013GRENV060. tel-01552806 HAL Id: tel-01552806 https://tel.archives-ouvertes.fr/tel-01552806 Submitted on 3 Jul 2017 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. THÈSE Pour obtenir le grade de DOCTEUR DE L’UNIVERSITÉ DE GRENOBLE Spécialité : Neurosciences - Neurobiologie Arrêté ministériel : 7 août 2006 Présentée par Erminia Fardone Thèse dirigée par Serge PEREZ codirigée par Alberto BRAVIN et Pantaleo ROMANELLI préparée au sein de l’ Installation européenne de rayonnement synchrotron (ESRF) dans l'École Doctorale Chimie et Sciences du Vivant A new application of microbeam radiation therapy (MRT) on the treatment of epilepsy and brain disorders Thèse soutenue publiquement le Vendredi 29 Novembre 2013, devant le jury composé de : Prof Alain Buisson (Président) Prof Université Joseph Fourier, Grenoble, France Prof Paola Coan (Examinateur) Prof Ludwig Maximilians University, Munich, Germany Prof Guido Cavaletti (Rapporteur) Prof University of Milan-Bicocca, Italy Prof Elisabeth Schültke (Rapporteur) MD, Dr Freiburg University Medical Center, Freiburg, Germany Dr Serge Perez (Directeur de Thèse) D.R. CNRS-CESMAV, Grenoble, France Dr Pantaleo Romanelli (Co-directeur de Thése) MD, Dr Brain Radiosurgery, Cyberknife Center, CDI Milan, Italy et Scientific Director, AB Medica, Italy Dr Alberto Bravin (Co-directeur de Thèse) Dr ESRF, Grenoble, France 1 Serendipity There's a big a big hard sun beaten on the big people in the big hard world Eddie Vedder – Hard Sun – 2 Acknowledgements This chapter of my life is completed; my new life in Grenoble and my PhD at the ESRF was made possible by many people around me. First of all I want to thank my jury, each member who accepted to evaluate this Thesis work and gave me the possibility to become a Philosophy doctor! I am grateful to Profs Elisabeth Schültke and Guido Cavaletti, for their precious time reading my thesis and for their constructive comments.I thank Profs Alain Buisson, Paola Coan and Silvio Garofalo for having accepted to be member of my jury. Furthermore, I want to thank the co-supervisor and main proposer of my PhD projects, Dr Pantaleo Romanelli, who, together with Dr Giuseppe Battaglia, gave me the opportunity to be a PhD student at the ESRF. I thank my co-supervisor at the ESRF Dr Alberto Bravin for his great teaching on how this Synchrotron works and then giving his support before and during these 3-year experiments. I am grateful to my Thesis Director Dr Serge Perez for the good advice and support specially during the last “hard” year of work. I thank Geraldine Le Duc for having hosted my animals, giving me the opportunity to create a new animal behavioural test never performed before in our team!!! I would like to thank Herwig Requardt for his support and patience during these 3 years of experiments! Furthermore, I am grateful to Elke Braur-Krisch and Christian Nemoz for their good advice, support and friendship. For many valuable encouragements and discussions on my analysis, I want to thank Domenico Bucci, Benoit Pouyatos and Lucia Ruocco that also shared with me their knowledge during our experiments. Each experiment wouldn’t have been possible without the useful help of Dominique Dallèry, Celine Leclec’H, Charlène Caloud and Hélène Bernard. Furthermore, I thank Thierry Brochard, Hélène Elleaume, Francois Estéve, Jeffrey Crosbie, Layal Obeid, “Trottinette” Gimenez and all ID17/INSERM-U836 Team 6 for having provided a stimulating and fun environment in which to learn and grow. Imma Martinez, Laura Nervo, Raph Serduc, Jeff Adam, Manu Brun, Elodie Riquet, Ludo Broche, FloFlo Taupin , Mél Flanders, “PiccolaStella” Potez, “Paulette “Fournier, Audrey Bouchet, Alberto Mittone and Loriane Weber provided encouragement, sound advice, good teaching, good company, and lots of good ideas to look at the future. A phone call or an e-mail during the writing days, helped me so much and brought a smile to my face especially when facing bed situations,. I would have been lost without them. I really thank you guys!!! A huge “thank you” to all the people that I met here in Grenoble, in particular Lory Dafne, Tommy, Ema, Mario, Ale, Lisa, Fede, Raffa, Franco Candido da Sant’Agapito, Vale, Isa, Valerio, Roberto, Sheeba e Davide, Paul, Bea , Massimo, Davide B., Carlotta and Yolanda. I cannot forget to mention my O’Callaghan friends: Gilou, Virgue, Flo and Stéphane . I thank you guys for your friendship and I will never forget our Guiness and our laughs! I wish to thank my entire family (my cousins, uncles and aunts) and my friends that from Isernia & Sant’Agapito creating a loving environment around me . Lastly, and most importantly, I wish to thank my parents, my sister and my grandma. They bore me, raised me, supported me, taught me, and loved me. To them I dedicate this Thesis. 3 Abstract A new application of microbeam radiation therapy (MRT) on the treatment of epilepsy and brain disorders Synchrotron-generated X-ray microplanar beams (microbeams, MBs) are characterized by the ability to avoid widespread tissue damage following delivery of doses ranging from hundreds to over a thousand of Grays. The resistance of normal tissues to high doses of MBs is likely related to the fast repair of the microvessels and to the wide interface between normal and irradiated tissue, allowing fast recolonization of the microbeam ablated columns of tissue. The preservation of the cortical architecture following high-dose microbeam irradiation and the ability to induce non-invasively the equivalent of a surgical cut over the cortex is of great interest for the development of novel experimental models in neurobiology and new treatment avenues for a variety of brain disorders. In this Thesis microbeams transections were delivered to the rat cortex and hippocampus. MBs cortical transections were delivered to the sensory motor cortex (size 100 µm/600 µm, center-to-center distance of 400 µm/1200 µm, peak-valley doses of 360-240 Gy/150-100 Gy) of Wistar male normal rats and rats developing seizures following cortical injections of Kainic Acid (KA). The motor performances following sensorimotor cortex transections were assessed by Rotarod tests. The effect of microbeam transections on cortical architecture was assessed by immunohistology with NeuN and GFAP, markers of mature neurons and astrocytes. No neurological deficit was observed in normal animals undergoing sensorimotor cortex microbeam transections. Convulsive seizure duration was markedly reduced following transections in KA rats. MBs hippocampal transections (75 µm thickness, 400 c-t-c distance and 600 Gy as peak entrance dose) were performed in adult normal Wistar rats. MBs transections were compared with a uniformly delivered X-ray dose (broad beam, BB) at 10 Gy. Our aims were to quantify (i) the impact of microbeams versus conventional irradiation on neurogenesis (using proliferative cells markers such as BrdU and Ki-67), and (ii) to investigate on a long term (1 year) the correlation between neurogenesis and impairments in learning and memory. Preservation of proliferative cells in the microbeam treated group was observed. Microbeam transections delivered to the hippocampus did not induce significant behavioural impairment histological and immunohistochemical findings showed a preserved hippocampal structure with evidence of highly precise transections parcellizing the hippocampus in columns. This work confirms the safe delivery of high doses of radiation to specific and radiosensitive parts of the brain, if performed using microbeams. The development of clinical devices delivering submillimetric beams able to generate cortical or hippocampal transections might become a new powerful new tool for the clinical treatment of epilepsy, and other functional brain disorders. Key words: Microbeam radiation therapy, radiosurgery, epilepsy, neurogenesis, brain ***************** Résumé Nouvelle application de la radiothérapie par microfaisceaux (MRT) pour le traitement de l’épilepsie et des troubles cérébraux Les microfaisceaux de rayons X générés par un synchrotron permettent de délivrer des doses de radiation très élevées, jusqu’à plusieurs centaines de gray (Gy), sans pour autant induire de 4 dommages tissulaires irréversibles dans les zones avoisinant la lésion. La réactivité du réseau vasculaire à se régénérer grâce aux cellules endothéliales, est probablement un mécanisme clef dans la radio-tolérance des tissus sains, puisqu’il permet une recolonisation rapide des zones tissulaires lésées. Cette méthode permet ainsi de reproduire de façon non invasive une incision chirurgicale précise du cortex tout en préservant son architecture. Cette caractéristique présente un intérêt certain et permet d’envisager le développement de nouveaux
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