New and Emerging fields in Bio-medicine Supporting Postgraduate Research with Internships in industry and Training Excellence SPRITE : Workpackage 1 New and Emerging fields in Bio-medicine Giovanna Muggiolu Dietrich Walsh Anne-Catherine Wéra

Workpackage 1: New and Emerging fields in Bio-medicine This Work Package is led by CNRS and aims to push the frontiers of biomedical application of ion beams, in particular it targets both the treatment of cancer as well as detectors for accurate dosimetry control. By linking each of the individual training plans with a clinical or industrial environment SPRITE will ensure that the “real world” objectives of the research are translated into the clinical environment for the benefit of patients. This Work Package is particularly timely, as treating cancer with ion beams (hadron therapy) is a rapidly developing field in Europe at the present time and there is an urgent need for individuals who have the multidisciplinary training in this field to take the technology forward.

BIO MEDECINE ? ESR Dietrich Walsh ER Anne-Catherine Wéra ESR UBW Munich, Germany ER Anne-Catherine Wéra University of Surrey Surrey, United Kingdom BIO MEDECINE ESR Giovanna Muggiolu CENBG/CNRS Bordeaux, France ESR Xxxx xxxxx CEA Caen, France ?

The use of charged particles for cancer therapy In 2011, the World Health Organisation recorded 57 millions death worldwide with 15% due to cancer. Main cancer treatments are: Surgery Chemotherapy Radiotherapy 50% of patient will undergo radiotherapy during the course of their treatment. > 95% of the case radiotherapy is carried on with X-rays >> Heavy charged particles Need for new therapeutic strategies

The use of charged particles for cancer therapy Traditional radiotherapy has side effects (i.e. chronic inflammation) Proton therapy advantageous due to Bragg peak Focussed proton beams may reduce side effects common in traditional radiotherapy (Work in progress) Carbon Therapy already in use in Japan (Gunma Heavy Ion Centre)

Targeted irradiation and comparison with broad beam irradiation of cells Workpackage 1: Micro Beam Allows the analysis of cellular damage response to highly localised areas Single particle – single cell irradiation Study of the hypersensitive response of lung tumour cells after low-dose irradiation with low LET protons

Targeted irradiation and comparison with broad beam irradiation of cells A549 cells irradiated with 4 MeV proton broad beam (10keV/µm) as/ar=14 ± 6.7 77% survival (HRS): RBE=10.7 ± 3.3 ( 10% survival: RBE= 1.9± 0.4) >> 10 times lower doses for the same effect End of HRS between 0.15 and 0.20 Gy

Targeted irradiation and comparison with broad beam irradiation of cells Micro Beam Irradiation A549 cells irradiated with 3.8 MeV proton micro beam (12keV/µm) as/ar=47 ± 14 77% survival (HRS): RBE=41 ± 10 End of HRS at 10 hits

>> Poisson Statistics? Targeted irradiation and comparison with broad beam irradiation of cells ? Threshold dose or Progressive transition ? >> Poisson Statistics?

End of the HRS between 0.15 – 0.20 Gy But, due to Poisson statistics Targeted irradiation and comparison with broad beam irradiation of cells End of the HRS between 0.15 – 0.20 Gy But, due to Poisson statistics 0.15 Gy: some cells will receive a dose higher than 0.20 Gy > Outside the HRS 0.20 Gy: some cells will receive a dose smaller than 0.15 Gy > Inside the HRS

(micro beam parameters) Targeted irradiation and comparison with broad beam irradiation of cells Broad beam surviving fraction extrapolation from micro beam data and Poisson statistics Poisson distribution Induced Repair Model (micro beam parameters) >> Poisson statistics can explain the variation in the threshold dose for HRS from broad beam to micro beam irradiation

Radio-sensitivity of sarcoma cell lines Sarcomas represent a heterogeneous group of rare tumours accounting for approximately 1% of adult cancers and with more than 50 histological subtypes Surgery is the primary approach to treating most sarcoma even though radiation therapy has also been used for those patients with residual tumours following surgery. Specific point mutations Specific chromosome translocations Genetic prospective Many limited amplifications Complex genomic profile

Centre d’Etudes Nucleaire Bordeaux-Gradignan Interdisciplinary research team from Physics, Chemistry and Biology IPCV Supervisor: H. Seznec Interdisciplinary Applications of Ions Beam in the Aquitaine Region AIFIRA

Radio-sensitivity of sarcoma cell lines Institute Bergonié Treatment Regional Cancer Centre Teaching Research Sarcoma team Unit INSERM U916 VINCO Leader Research: F. Chibon 40 sarcoma cell lines from patients which are highly-well characterized from genetic/genomic and phenotypic point of view.

Radio-sensitivity of sarcoma cell lines Sarcomas lines well-characterized IB 115 amplifications of MDM2 and CDK4 genes IB 106 unclassified sarcomas from paravertebral mass with heterogenic pleomorphic cells

Radio-sensitivity of sarcoma cell lines GOAL Evaluate the radiation sensitivity of sarcoma cell lines Are sarcoma cells sensitive to photon therapy? Are they sensitive also to proton therapy? Have these two kind of radiations the same efficacy? Comparing the radiation sensitivity of these cell lines using a photon facility (normally used for patients’ treatment) and a proton micro-beam (useful to study the proton-therapy)

Radio-sensitivity of sarcoma cell lines EXPECTATIONS SHORT TIME Selection of Sarcoma cell lines according to their genetic point of view Definition of an irradiation protocol from broad beam to proton beam (define irradiation mode, irradiation dose and radiation types) Selection of biological end-points Comparison Proton vs X-rays irradiation LONG TIME Development and validation of in vitro irradiation protocols combining both protons and nanoparticles

Role of mitochondria in the cellular response to ionising radiation What are Mitochondria? Mitochondria are biological „powerplants“. There are 1-1000 per cell and they range from 500nm-2µm Main source of energy in a cell (Energy = ATP) More energy a cell requires, more mitochondria it has Cells can not survive without mitochondria! Mitochondria have a double membrane which they use to keep a constant ΔV between mitochondria and inside of the cell (polarisation). This is required for function Mitochondria have their own DNA which is inherited from the mother.

Role of mitochondria in the cellular response to ionising radiation Roles of the Mitochondria in cells Mitochondria are the energy factories of cells Use sugar to produce cellular energy (ATP) the process requires oxygen and mitochondria (Slow rate, high ATP yield) Mitochondria regulate energy „metabolism“ of cells Cancer cells require high amounts of energy Mitochondrial disfunction is involved in many diseases

Role of mitochondria in the cellular response to ionising radiation To identify biological effects of microbeam irradiation on Mitochondria. Initial Ideas: Microbeam irradiation of Mitochondria in live cells (novel techniques employed) Live cell imaging of mitochondrial depolarisation (Does pinpoint radiation destroy mitochondria?) Analysis of gene expression level of mitochondrial proteins (qRT-PCR) Imaging of DNA strand breaks in mitochondrial DNA

Role of mitochondria in the cellular response to ionising radiation SNAKE Superconducting nanoprobe for applied nuclear physics experiments Up to 25 MeV protons Lateral resolution of 1µm

Summary Diverse projects in the field of radiation biology and biomedicine The goal is to bring together physicists and biologist in a clinical and industrial environment The research being conducted aims to answer emerging questions in the field of experimental radiation oncology which are relevant for future particle therapy.

THANK YOU FOR YOUR ATTENTION!!! SPRITE : Workpackage 1 New and Emerging fields in Bio-medicine Giovanna Muggiolu Dietrich Walsh Anne-Catherine Wéra